Differential Production of Phenolics, Lipids, Carbohydrates and Proteins in Stressed and Unstressed Aquatic Plants, Azolla filiculoides and Azolla pinnata.

The metabolic plasticity of shikimate and phenylpropanoid pathways redirects carbon flow to different sink products in order to protect sessile plants from environmental stresses. This study assessed the biochemical responses of two Azolla species, A. filiculoides and A. pinnata, to the combined effects of environmental and nutritional stresses experienced while growing outdoors under Australian summer conditions. These stresses triggered a more than 2-fold increase in the production of total phenols and their representatives, anthocyanins (up to 18-fold), flavonoids (up to 4.7-fold), and condensed tannins (up to 2.7-fold), which led to intense red coloration of the leaves. These changes were also associated with an increase in the concentration of carbohydrates and a decrease in concentrations of lipids and total proteins. Changes in lipid biosynthesis did not cause significant changes in concentrations of palmitoleic acid (C16:0), linolenic acid (C18:3), and linoleic acid (C18:2), the fatty acid signatures of Azolla species. However, a reduction in protein production triggered changes in biosynthesis of alanine, arginine, leucine, tyrosine, threonine, valine, and methionine amino acids. Stress-triggered changes in key nutritional components, phenolics, lipids, proteins, and carbohydrates could have a significant impact on the nutritional value of both Azolla species, which are widely used as a sustainable food supplement for livestock, poultry, and fish industries.

Population- and sex-specific sensitivity of the marine amphipod Allorchestes compressa to metal exposure.

The sensitivity to contaminants of natural populations varies greatly depending on their historical exposure and on the sex of the individual. These factors result in great uncertainty in ecotoxicological risk assessments and challenge the protection of marine biodiversity. This study investigated the role of background pollution in the environment in shaping the sensitivity of males and females of the common marine amphipod Allorchestes compressa to the common trace marine pollutant, copper (Cu). Female and male amphipods were collected from two sites: Geelong (the polluted site) and Clifton Springs (the clean site). Amphipods were exposed to Cu treatments of 0, 50, 100, and 250 µg/L for 10 days, followed by a 10-day recovery period. Cu-exposed males from Geelong showed a reduction in feeding rate at a higher Cu concentration than males from Clifton Springs, suggesting that they have a higher tolerance to Cu than males from Clifton Springs. This can be explained by their higher base level of metallothioneins (MTs) and glutathione-S-transferase (GST), the key physiological responses for detoxification and defence against damages from Cu toxicity. Males showed a higher tolerance to Cu than females. This pattern was similar in both populations, which may be associated with a higher level of GST. During the recovery period, only males from Geelong fully recovered to the control level. Our results emphasize the importance of considering population- and sex-specific sensitivity of invertebrates to contaminants in ecotoxicological risk assessments.

Physicochemical Characteristics of Protein Isolated from Thraustochytrid Oilcake.

The oil from thraustochytrids, unicellular heterotrophic marine protists, is increasingly used in the food and biotechnological industries as it is rich in omega-3 fatty acids, squalene and a broad spectrum of carotenoids. This study showed that the oilcake, a by-product of oil extraction, is equally valuable as it contained 38% protein/dry mass, and thraustochytrid protein isolate can be obtained with 92% protein content and recovered with 70% efficiency. The highest and lowest solubilities of proteins were observed at pH 12.0 and 4.0, respectively, the latter being its isoelectric point. Aspartic acid, glutamic acid, histidine, and arginine were the most abundant amino acids in proteins. The arginine-to-lysine ratio was higher than one, which is desired in heart-healthy foods. The denaturation temperature of proteins ranged from 167.8-174.5 °C, indicating its high thermal stability. Proteins also showed high emulsion activity (784.1 m2/g) and emulsion stability (209.9 min) indices. The extracted omega-3-rich oil melted in the range of 30-34.6 °C and remained stable up to 163-213 °C. This study shows that thraustochytrids are not only a valuable source of omega 3-, squalene- and carotenoid-containing oils, but are also rich in high-value protein with characteristics similar to those from oilseeds.

Marine Protists and Rhodotorula Yeast as Bio-Convertors of Marine Waste into Nutrient-Rich Deposits for Mangrove Ecosystems.

This paper represents a comprehensive study of two new thraustochytrids and a marine Rhodotorula red yeast isolated from Australian coastal waters for their abilities to be a potential renewable feedstock for the nutraceutical, food, fishery and bioenergy industries. Mixotrophic growth of these species was assessed in the presence of different carbon sources: glycerol, glucose, fructose, galactose, xylose, and sucrose, starch, cellulose, malt extract, and potato peels. Up to 14g DW/L (4.6gDW/L-day and 2.8gDW/L-day) of biomass were produced by Aurantiochytrium and Thraustochytrium species, respectively. Thraustochytrids biomass contained up to 33% DW of lipids, rich in omega-3 polyunsaturated docosahexaenoic acid (C22:6, 124mg/g DW); up to 10.2mg/gDW of squalene and up to 61µg/gDW of total carotenoids, composed of astaxanthin, canthaxanthin, echinenone, and ß-carotene. Along with the accumulation of these added-value chemicals in biomass, thraustochytrid representatives showed the ability to secrete extracellular polysaccharide matrixes containing lipids and proteins. Rhodotorula sp lipids (26% DW) were enriched in palmitic acid (C16:0, 18mg/gDW) and oleic acid (C18:1, 41mg/gDW). Carotenoids (87µg/gDW) were mainly represented by ß-carotene (up to 54µg/gDW). Efficient growth on organic and inorganic sources of carbon and nitrogen from natural and anthropogenic wastewater pollutants along with intracellular and extracellular production of valuable nutrients makes the production of valuable chemicals from isolated species economical and sustainable.

Aquatic Plants, Landoltia punctata, and Azolla filiculoides as Bio-Converters of Wastewater to Biofuel.

The aquatic plants, Azolla filiculoides, and Landoltia punctate, were used as complementing phytoremediators of wastewater containing high levels of phosphate, which simulates the effluents from textile, dyeing, and laundry detergent industries. Their complementarities are based on differences in capacities to uptake nitrogen and phosphate components from wastewater. Sequential treatment by L. punctata followed by A. filiculoides led to complete removal of NH4, NO3, and up to 93% reduction of PO4. In experiments where L. punctata treatment was followed by fresh L. punctata, PO4 concentration was reduced by 65%. The toxicity of wastewater assessed by shrimps, Paratya australiensis, showed a four-fold reduction of their mortality (LC50 value) after treatment. Collected dry biomass was used as an alternative carbon source for heterotrophic marine protists, thraustochytrids, which produced up to 35% dry weight of lipids rich in palmitic acid (50% of total fatty acids), the key fatty acid for biodiesel production. The fermentation of treated L. punctata biomass by Enterobacter cloacae yielded up to 2.14 mol H2/mole of reduced sugar, which is comparable with leading terrestrial feedstocks. A. filiculoides and L. punctata can be used as a new generation of feedstock, which can treat different types of wastewater and represent renewable and sustainable feedstock for bioenergy production.

The Nutritional and Pharmacological Potential of New Australian Thraustochytrids Isolated from Mangrove Sediments.

Mangrove sediments represent unique microbial ecosystems that act as a buffer zone, biogeochemically recycling marine waste into nutrient-rich depositions for marine and terrestrial species. Marine unicellular protists, thraustochytrids, colonizing mangrove sediments have received attention due to their ability to produce large amounts of long-chain ω3-polyunsaturated fatty acids. This paper represents a comprehensive study of two new thraustochytrids for their production of valuable biomolecules in biomass, de-oiled cakes, supernatants, extracellular polysaccharide matrixes, and recovered oil bodies. Extracted lipids (up to 40% of DW) rich in polyunsaturated fatty acids (up to 80% of total fatty acids) were mainly represented by docosahexaenoic acid (75% of polyunsaturated fatty acids). Cells also showed accumulation of squalene (up to 13 mg/g DW) and carotenoids (up to 72 µg/g DW represented by astaxanthin, canthaxanthin, echinenone, and ß-carotene). Both strains showed a high concentration of protein in biomass (29% DW) and supernatants (2.7 g/L) as part of extracellular polysaccharide matrixes. Alkalinization of collected biomass represents a new and easy way to recover lipid-rich oil bodies in the form of an aqueous emulsion. The ability to produce added-value molecules makes thraustochytrids an important alternative to microalgae and plants dominating in the food, pharmacological, nutraceutical, and cosmetics industries.

Effects of perfluorooctanoic acid (PFOA) on the thyroid status, vitellogenin, and oxidant-antioxidant balance in the Murray River rainbowfish.

Perfluorooctanoic acid's (PFOA) widespread use, presence and persistence in the aquatic environment has led to an increasing number of studies focusing on its toxicological effects. In Australia, PFOA has been detected in the aquatic environment, however its effects on Australian native fauna are unknown. In this study, male Australian native fish Murray River rainbowfish (Melanotaenia fluviatilis) were exposed to four different concentrations of PFOA (0.01, 0.1, 1 and 10 mg L-1). Variations in thyroid hormones (Triiodothyronine (T3)/Thyroxine (T4)) and the presence of vitellogenin were determined in plasma. Oxidative stress responses were evaluated in gills and liver. Exposure of male fish to PFOA resulted in altered T3/T4 ratios and the presence of vitellogenin in the plasma. Activities of catalase (CAT) and glutathione- S-transferase (GST) were significantly increased in the gills and significantly reduced in the liver. Lipid peroxidation was observed in both tissues showing that vital organs could not neutralize the peroxides generated by oxidative stress resulting from exposure to PFOA. In natural populations exposed to PFOA, such hormonal disturbances can have negative effects, notably through altered capacity to respond to changes in environmental conditions.

The Toxicity of Nonaged and Aged Coated Silver Nanoparticles to Freshwater Alga Raphidocelis subcapitata.

The transformation of coated silver nanoparticles (AgNPs) and their impacts on aquatic organisms require further study. The present study investigated the role of aging on the transformation of differently coated AgNPs and their sublethal effects on the freshwater alga Raphidocelis subcapitata. The stability of AgNPs was evaluated over 32 d, and the results indicated that transformation of AgNPs occurred during the incubation; however, coating-specific effects were observed. Fresh AgNPs increased reactive oxygen species (ROS) formation, whereas aged AgNPs induced excessive ROS generation compared with their fresh counterparts. Increased ROS levels caused increased lipid peroxidation (LPO) in treatment groups exposed to both fresh and aged NPs, although LPO was comparatively higher in algae exposed to aged AgNPs. The observed increase in catalase (CAT) activity of algal cells was attributed to early stress responses induced by excessive intracellular ROS generation, and CAT levels were higher in the aged NP treatment groups. In conclusion, AgNPs increased ROS levels and LPO in algae and caused the activation of antioxidant enzymes such as CAT. Overall, the results suggest that aging and coating of AgNPs have major impacts on AgNP transformation in media and their effects on algae. Environ Toxicol Chem 2019;38:2371-2382. © 2019 SETAC.

The toxicity of coated silver nanoparticles to Daphnia carinata and trophic transfer from alga Raphidocelis subcapitata.

Nanoparticles (NPs) are causing threats to the environment. Silver NPs (AgNPs) are increasingly used in commercial products and may end up in freshwater ecosystems. The freshwater organisms are vulnerable due to water-borne and dietary exposure to AgNPs. Surface properties play an important role in the fate and behavior of AgNPs in the aquatic environment and their effects on organisms. However, effects of surface properties of AgNPs on organisms are poorly understood. In this study, we explored the effects of AgNPs coated with three different ligands; Tyrosine (T-AgNP), Epigallocatechin gallate (E-AgNP) and Curcumin (C-AgNP) in relation to the toxicity to a key aquatic organism; Daphnia carinata. The study focused on how coatings determine fate of NPs in the medium, mortality, feeding behaviour, bioaccumulation and trophic transfer from the freshwater alga, Raphidocelis subcapitata to daphnids. NP stability tests indicated that T-AgNPs were least stable in the ASTM daphnia medium while C-AgNPs were most stable. 48 h EC50 values of AgNPs to D. carinata were in the order of E-AgNP (19.37 µg L-1) > C-AgNP (21.37 µg L-1) > T-AgNP (49.74 µg L-1) while the 48 h EC50 value of Ag+ ions was 1.21 µg L-1. AgNP contaminated algae significantly decreased the feeding rates of daphnids. However, no significant differences were observed in feeding rates between algae contaminated with differently coated AgNPs. Trophic transfer studies showed that AgNPs were transferred from algae to daphnids. The bioacumulation of AgNPs in algae and the diet-borne bioaccumulation of AgNPs in daphnids varied for differently coated AgNPs. Bioaccumulation of C-AgNPs in algae was 1.5 time higher than T-AgNPs. However, the accumulation of T-AgNPs in daphnids via trophic transfer was 2.6 times higher than T-AgNPs. The knowledge generated from this study enhances the understanding of surface property dependent toxicity, bioaccumulation and trophic transfer of AgNPs in aquatic environments.

The toxicity of non-aged and aged coated silver nanoparticles to the freshwater shrimp Paratya australiensis.

Nanoparticles (NPs) transform in the environment which result in alterations to their physicochemical properties. However, the effects of aging on the toxicity of NPs to aquatic organisms remain to be determined. Further the reports that have been published present contradictory results. The aim of this study was to examine the stability of differently coated silver nanoparticles (AgNPs) in media and the influence of aging of these NP on potential toxicity to freshwater shrimp Paratya australiensis. Coating-dependent changes in the stability of AgNP were observed with aging. Curcumin (C) coated AgNPs were stable, while tyrosine (T) coated AgNPs and epigallocatechin gallate (E) coated AgNPs aggregated in the P. australiensis medium. Increased lipid peroxidation and catalase activity was noted in P. australiensis exposed to AgNPs, suggesting oxidative stress was associated with NP exposure. The enhanced oxidative stress initiated by aged C-AgNPs suggests that aging of these NPs produced different toxicological responses. In summary, data suggest that coating-dependent alterations in NPs, together with aging affect both persistence and subsequent toxicity of NPs to freshwater organisms. Thus, the coating-dependent fate and toxicity of AgNPs together with the effect of their aging need to be considered in assessing the environmental risk of AgNPs to aquatic organisms.

Lipid production in aquatic plant Azolla at vegetative and reproductive stages and in response to abiotic stress.

The aquatic plant Azolla became increasingly popular as bioenergy feedstock because of its high growth rate, production of biomass with high levels of biofuel-producing molecules and ability to grow on marginal lands. In this study, we analysed the contribution of all organs of Azolla to the total yield of lipids at vegetative and reproductive stages and in response to stress. Triacylglycerol-containing lipid droplets were detected in all (vegetative and reproductive) organs with the highest level in the male microsporocarps and microspores. As a result, significantly higher total yields of lipids were detected in Azolla filiculoides and Azolla pinnata at the reproductive stage. Starving changed the yield and composition of the fatty acid as a result of re-direction of carbon flow from fatty acid to anthocyanin pathways. The composition of lipids, in regard the length and degree of unsaturation of fatty acids, in Azolla meets most of the important requirements for biodiesel standards. The ability of Azolla to grow on wastewaters, along with their high productivity rate, makes it an attractive feedstock for the production of biofuels.

New Insights in Vanishing White Matter Disease: Isolated Bilateral Optic Neuropathy in Adult Onset Disease.

BACKGROUND: Vanishing white matter disease (VWMD) is a rare disease affecting cerebral white matter. The adult form is even rarer and manifests with motor symptoms, behavioral problems, and dementia. There is no treatment and progression is inevitable. We describe a case with atypical manifestations and an unusual course. METHODS: Description of a 42-year-old man with VWMD complaining of progressive visual loss in the right eye. RESULTS: The patient's visual acuity was 20/60, right eye, and 20/25, left eye, with pale optic nerves bilaterally. MRI showed atrophy of the corpus callosum, diffuse rarefaction of cerebral white matter including the anterior and posterior visual pathways. CONCLUSION: Our patient had no further symptoms besides loss of visual acuity, which is rare in patients with VWMD of the same age and genetic mutation.

Applications of microalgal biofilms for wastewater treatment and bioenergy production.

BACKGROUND: Microalgae have shown clear advantages for the production of biofuels compared with energy crops. Apart from their high growth rates and substantial lipid/triacylglycerol yields, microalgae can grow in wastewaters (animal, municipal and mining wastewaters) efficiently removing their primary nutrients (C, N, and P), heavy metals and micropollutants, and they do not compete with crops for arable lands. However, fundamental barriers to the industrial application of microalgae for biofuel production still include high costs of removing the algae from the water and the water from the algae which can account for up to 30-40% of the total cost of biodiesel production. Algal biofilms are becoming increasingly popular as a strategy for the concentration of microalgae, making harvesting/dewatering easier and cheaper. RESULTS: We have isolated and characterized a number of natural microalgal biofilms from freshwater, saline lakes and marine habitats. Structurally, these biofilms represent complex consortia of unicellular and multicellular, photosynthetic and heterotrophic inhabitants, such as cyanobacteria, microalgae, diatoms, bacteria, and fungi. Biofilm #52 was used as feedstock for bioenergy production. Dark fermentation of its biomass by Enterobacter cloacae DT-1 led to the production of 2.4 mol of H2/mol of reduced sugar. The levels and compositions of saturated, monosaturated and polyunsaturated fatty acids in Biofilm #52 were target-wise modified through the promotion of the growth of selected individual photosynthetic inhabitants. Photosynthetic components isolated from different biofilms were used for tailoring of novel biofilms designed for (i) treatment of specific types of wastewaters, such as reverse osmosis concentrate, (ii) compositions of total fatty acids with a new degree of unsaturation and (iii) bio-flocculation and concentration of commercial microalgal cells. Treatment of different types of wastewaters with biofilms showed a reduction in the concentrations of key nutrients, such as phosphates, ammonia, nitrates, selenium and heavy metals. CONCLUSIONS: This multidisciplinary study showed the new potential of natural biofilms, their individual photosynthetic inhabitants and assembled new algal/cyanobacterial biofilms as the next generation of bioenergy feedstocks which can grow using wastewaters as a cheap source of key nutrients.

Horizontal Gaze Palsy and Progressive Scoliosis With ROBO 3 Mutations in Patients From Cape Verde.

Horizontal gaze palsy with progressive scoliosis (HGPPS) is a rare and autosomal recessive syndrome. We describe 2 cases of HGPPS which are the first documented in patients of African ancestry from an isolated population in Cape Verde. They demonstrated typical findings on neuro-ophthalmic examination and brain magnetic resonance imaging. One patient had novel heterozymous mutations of the ROB0 3 gene.

Aquatic plant Azolla as the universal feedstock for biofuel production.

BACKGROUND: The quest for sustainable production of renewable and cheap biofuels has triggered an intensive search for domestication of the next generation of bioenergy crops. Aquatic plants which can rapidly colonize wetlands are attracting attention because of their ability to grow in wastewaters and produce large amounts of biomass. Representatives of Azolla species are some of the fastest growing plants, producing substantial biomass when growing in contaminated water and natural ecosystems. Together with their evolutional symbiont, the cyanobacterium Anabaena azollae, Azolla biomass has a unique chemical composition accumulating in each leaf including three major types of bioenergy molecules: cellulose/hemicellulose, starch and lipids, resembling combinations of terrestrial bioenergy crops and microalgae. RESULTS: The growth of Azolla filiculoides in synthetic wastewater led up to 25, 69, 24 and 40 % reduction of NH4-N, NO3-N, PO4-P and selenium, respectively, after 5 days of treatment. This led to a 2.6-fold reduction in toxicity of the treated wastewater to shrimps, common inhabitants of wetlands. Two Azolla species, Azolla filiculoides and Azolla pinnata, were used as feedstock for the production of a range of functional hydrocarbons through hydrothermal liquefaction, bio-hydrogen and bio-ethanol. Given the high annual productivity of Azolla, hydrothermal liquefaction can lead to the theoretical production of 20.2 t/ha-year of bio-oil and 48 t/ha-year of bio-char. The ethanol production from Azolla filiculoides, 11.7 × 103 L/ha-year, is close to that from corn stover (13.3 × 103 L/ha-year), but higher than from miscanthus (2.3 × 103 L/ha-year) and woody plants, such as willow (0.3 × 103 L/ha-year) and poplar (1.3 × 103 L/ha-year). With a high C/N ratio, fermentation of Azolla biomass generates 2.2 mol/mol glucose/xylose of hydrogen, making this species a competitive feedstock for hydrogen production compared with other bioenergy crops. CONCLUSIONS: The high productivity, the ability to grow on wastewaters and unique chemical composition make Azolla species the most attractive, sustainable and universal feedstock for low cost, low energy demanding, near zero maintenance system for the production of a wide spectrum of renewable biofuels.

OUTCOMES AFTER PARS PLANA VITRECTOMY FOR EPIRETINAL MEMBRANES ASSOCIATED WITH TOXOPLASMOSIS.

PURPOSE: To evaluate outcomes and complications of pars plana vitrectomy in patients with epiretinal membrane secondary to toxoplasmic retinochoroiditis. METHODS: Retrospective evaluation of the records of 14 patients who underwent pars plana vitrectomy for epiretinal membrane secondary to toxoplasmic retinochoroiditis. The best-corrected visual acuity, intraoperative and postoperative complications, and macular optical coherence tomography were analysed. All patients received postoperative prophylactic treatment with trimethoprim/sulfamethoxazole. RESULTS: Fourteen patients, 5 men and 9 women, were included. Mean follow-up period after surgery was 6.07 ± 2.64 months. Preoperative mean best-corrected visual acuity was 20/200, and postoperative mean best-corrected visual acuity was 20/60. There were no intraoperative complications. Three patients developed posterior capsule opacification, and one patient developed cataract. CONCLUSION: Pars plana vitrectomy is a safe and effective procedure in patients with epiretinal membrane secondary to toxoplasmic retinochoroiditis, improving both visual acuity and anatomical result on macular optical coherence tomography. The most frequent postoperative complications were posterior capsule opacification and cataract. No recurrences of the disease were recorded.

Lipid production in association of filamentous fungi with genetically modified cyanobacterial cells.

BACKGROUND: Numerous strategies have evolved recently for the generation of genetically modified or synthetic microalgae and cyanobacteria designed for production of ethanol, biodiesel and other fuels. In spite of their obvious attractiveness there are still a number of challenges that can affect their economic viability: the high costs associated with (1) harvesting, which can account for up to 50 % of the total biofuel's cost, (2) nutrients supply and (3) oil extraction. Fungal-assisted bio-flocculation of microalgae is gaining increasing attention due to its high efficiency, no need for added chemicals and low energy inputs. The implementation of renewable alternative carbon, nitrogen and phosphorus sources from agricultural wastes and wastewaters for growing algae and fungi makes this strategy economically attractive. RESULTS: This work demonstrates that the filamentous fungi, Aspergillus fumigatus can efficiently flocculate the unicellular cyanobacteria Synechocystis PCC 6803 and its genetically modified derivatives that have been altered to enable secretion of free fatty acids into growth media. Secreted free fatty acids are potentially used by fungal cells as a carbon source for growth and ex-novo production of lipids. For most of genetically modified strains the total lipid yields extracted from the fungal-cyanobacterial pellets were found to be higher than additive yields of lipids and total free fatty acids produced by fungal and Synechocystis components when grown in mono-cultures. The synergistic effect observed in fungal-Synechocystis associations was also found in bioremediation rates when animal husbandry wastewater was used an alternative source of nitrogen and phosphorus. CONCLUSION: Fungal assisted flocculation can complement and assist in large scale biofuel production from wild-type and genetically modified Synechocystis PCC 6803 strains by (1) efficient harvesting of cyanobacterial cells and (2) producing of high yields of lipids accumulated in fungal-cyanobacterial pellets.

Evaluating the efficacy of bioremediating a diesel-contaminated soil using ecotoxicological and bacterial community indices.

Diesel represents a common environmental contaminant as a result of operation, storage, and transportation accidents. The bioremediation of diesel in a contaminated soil is seen as an environmentally safe approach to treat contaminated land. The effectiveness of the remediation process is usually assessed by the degradation of the total petroleum hydrocarbon (TPH) concentration, without considering ecotoxicological effects. The aim of this study was to assess the efficacy of two bioremediation strategies in terms of reduction in TPH concentration together with ecotoxicity indices and changes in the bacterial diversity assessed using PCR-denaturing gradient gel electrophoresis (DGGE). The biostimulation strategy resulted in a 90 % reduction in the TPH concentration versus 78 % reduction from the natural attenuation strategy over 12 weeks incubation in a laboratory mesocosm-containing diesel-contaminated soil. In contrast, the reduction in the ecotoxicity resulting from the natural attenuation treatment using the Microtox and earthworm toxicity assays was more than double the reduction resulting from the biostimulation treatment (45 and 20 % reduction, respectively). The biostimulated treatment involved the addition of nitrogen and phosphorus in order to stimulate the microorganisms by creating an optimal C:N:P molar ratio. An increased concentration of ammonium and phosphate was detected in the biostimulated soil compared with the naturally attenuated samples before and after the remediation process. Furthermore, through PCR-DGGE, significant changes in the bacterial community were observed as a consequence of adding the nutrients together with the diesel (biostimulation), resulting in the formation of distinctly different bacterial communities in the soil subjected to the two strategies used in this study. These findings indicate the suitability of both bioremediation approaches in treating hydrocarbon-contaminated soil, particularly biostimulation. Although biostimulation represents a commercially viable bioremediation technology for use in diesel-contaminated soils, further research is required to determine the ecotoxicological impacts of the intervention.

Fungal-assisted algal flocculation: application in wastewater treatment and biofuel production.

BACKGROUND: The microalgal-based industries are facing a number of important challenges that in turn affect their economic viability. Arguably the most important of these are associated with the high costs of harvesting and dewatering of the microalgal cells, the costs and sustainability of nutrient supplies and costly methods for large scale oil extraction. Existing harvesting technologies, which can account for up to 50% of the total cost, are not economically feasible because of either requiring too much energy or the addition of chemicals. Fungal-assisted flocculation is currently receiving increased attention because of its high harvesting efficiency. Moreover, some of fungal and microalgal strains are well known for their ability to treat wastewater, generating biomass which represents a renewable and sustainable feedstock for bioenergy production. RESULTS: We screened 33 fungal strains, isolated from compost, straws and soil for their lipid content and flocculation efficiencies against representatives of microalgae commercially used for biodiesel production, namely the heterotrophic freshwater microalgae Chlorella protothecoides and the marine microalgae Tetraselmis suecica. Lipid levels and composition were analyzed in fungal-algal pellets grown on media containing alternative carbon, nitrogen and phosphorus sources from wheat straw and swine wastewater, respectively. The biomass of fungal-algal pellets grown on swine wastewater was used as feedstock for the production of value-added chemicals, biogas, bio-solids and liquid petrochemicals through pyrolysis. Co-cultivation of microalgae and filamentous fungus increased total biomass production, lipid yield and wastewater bioremediation efficiency. CONCLUSION: Fungal-assisted microalgal flocculation shows significant potential for solving the major challenges facing the commercialization of microalgal biotechnology, namely (i) the efficient and cost-effective harvesting of freshwater and seawater algal strains; (ii) enhancement of total oil production and optimization of its composition; (iii) nutrient supply through recovering of the primary nutrients, nitrogen and phosphates and microelements from wastewater. The biomass generated was thermochemically converted into biogas, bio-solids and a range of liquid petrochemicals including straight-chain C12 to C21 alkanes which can be directly used as a glycerine-free component of biodiesel. Pyrolysis represents an efficient alternative strategy for biofuel production from species with tough cell walls such as fungi and fungal-algal pellets.

Co-cultivation of fungal and microalgal cells as an efficient system for harvesting microalgal cells, lipid production and wastewater treatment.

The challenges which the large scale microalgal industry is facing are associated with the high cost of key operations such as harvesting, nutrient supply and oil extraction. The high-energy input for harvesting makes current commercial microalgal biodiesel production economically unfeasible and can account for up to 50% of the total cost of biofuel production. Co-cultivation of fungal and microalgal cells is getting increasing attention because of high efficiency of bio-flocculation of microalgal cells with no requirement for added chemicals and low energy inputs. Moreover, some fungal and microalgal strains are well known for their exceptional ability to purify wastewater, generating biomass that represents a renewable and sustainable feedstock for biofuel production. We have screened the flocculation efficiency of the filamentous fungus A. fumigatus against 11 microalgae representing freshwater, marine, small (5 µm), large (over 300 µm), heterotrophic, photoautotrophic, motile and non-motile strains. Some of the strains are commercially used for biofuel production. Lipid production and composition were analysed in fungal-algal pellets grown on media containing alternative carbon, nitrogen and phosphorus sources contained in wheat straw and swine wastewater, respectively. Co-cultivation of algae and A. fumigatus cells showed additive and synergistic effects on biomass production, lipid yield and wastewater bioremediation efficiency. Analysis of fungal-algal pellet's fatty acids composition suggested that it can be tailored and optimised through co-cultivating different algae and fungi without the need for genetic modification.