Toxic effects of pentachlorophenol, azinphos-methyl and chlorpyrifos on the development of Paracentrotus lividus embryos.

The application of many current-use pesticides has increased after the disuse of persistent, bioaccumulative or toxic ones as DDT or chlordane. Many of the used pesticides are considered less dangerous towards the environment for their physico-chemical properties. This study investigated the toxic effects of three current-use pesticides, pentachlorophenol (PCP), azinphos-methyl (AZM), and chlorpyrifos, on Mediterranean sea urchin Paracentrotus lividus early development and offspring quality. The experimental results showed that the most toxic pesticides were PCP and AZM at EC50 level. Nevertheless at low concentration PCP resulted the less toxic compound and showed EC1 value more protective than NOEC. PCP at high concentration seemed to modify cytoskeleton assembly, while at low concentrations, it could alter the deposition of the larval skeleton. OPs at low concentrations until 300 µg/l showed a similar toxicological behaviour with a trend corresponding to the pesticide concentrations. At high concentration (500 µg/l) the effect mainly observed was the embryos pre-larval arrest. This investigation highlighted the relevance to evaluate, in coastal seawaters, the levels of the used pesticides to understand the real impact on benthic populations mainly in sites characterized by intensive agriculture or floriculture activities, such as the coastal areas of the Mediterranean Sea.

Cadmium, lead and their mixtures with copper: Paracentrotus lividus embryotoxicity assessment, prediction, and offspring quality evaluation.

The aim of this research was to assess the combined effects of three heavy metals (copper, lead, cadmium) on the fertilization and offspring quality of the sea urchin Paracentrotus lividus at EC50, NOEL, and EC1 concentrations. The observed data were compared with the predictions derived from approaches of Concentration Addition (CA) and Independent Action (IA) in order to evaluate the proper prediction of the observed mixture toxic effect. The P. lividus embryotoxicity of trace metals decreases as follows: Cu > Pb > Cd at all toxicity concentration tested. EC50 mixture revealed less toxic only than Cu; EC50 was 0.80 (± 0.07) mg/l, the offspring malformations were mainly P1 type (skeletal alterations) up to 20% mixture concentration, and P2 type from 70% concentration. The NOEL and EC1 mixtures evidenced that all compounds contribute to the overall toxicity, even if present at low concentrations: the former EC50 was 0.532 (± 0.058) mg/l and the latter was 1.081 (± 0.240) mg/l. The developmental defects observed were mainly P1 type in both mixtures. Both CA and IA models did not accurately predict mixture toxicity for EC50 and NOEL mixtures. Instead, EC1 mixture effects seemed well represented by the IA model. The protective action of the CA model, although quite accurate when applied to simple biological systems like algae and bacteria, but failed to represent the worst-case in this study with more complex organisms. It would be useful to introduce in the models one or more factors that take into account the complexity of these biological systems.

Biochemical composition and in vitro digestibility of Galdieria sulphuraria grown on spent cherry-brine liquid.

The aim of this work was to valorise an industrial food by-product and to produce a microalgal biomass rich in phytochemicals at high added value for food and nutraceutical applications. The biochemical composition, in vitro digestibility and antioxidant activity of Galdieria sulphuraria biomass grown heterotrophically on standard medium (SM) and on spent Cherry-Brine Liquid (sCBL) were assessed and compared. The biomass produced in sCBL was characterized by a lower content of proteins and lipids, while showing an increase in carbohydrates and polyphenols (5.3 vs 1.6 mg g-1). The sCBL biomass lipid moiety had a lower palmitic and linoleic acid content and a higher oleic acid concentration than SM. The total protein digestibility of Galdieria grown in SM and sCBL was 79% and 63% respectively. The antioxidant activity (AA) of G. sulphuraria biomass grown in sCBL was significantly higher than that grown in SM. Studying the AA release for sCBL biomass during the digestion, the highest value was found in the intestinal phase. In conclusion, G. sulphuraria has a valuable nutritional profile and could become a valuable source of phytochemicals, depending on the cultivation media. Cultivation on sCBL would allow an environmentally and economically sustainable process, valorising the food by-product and producing a microalgal biomass rich in cherry anthocyanins with high AA released at the intestinal level.

Evaluation of anaerobic digestates from different feedstocks as growth media for Tetradesmus obliquus, Botryococcus braunii, Phaeodactylum tricornutum and Arthrospira maxima.

In this paper, two freshwater microalgae (Tetradesmus obliquus and Botryococcus braunii), a marine diatom (Phaeodactylum tricornutum) and a photosynthetic cyanobacterium (Arthrospira maxima) were investigated for their ability to grow on liquid digestates (LDs). Three LDs were obtained from anaerobic digestion of different organic wastes: zootechnical (ZW LD), vegetable biomass (VW LD) and the organic fraction of municipal solid wastes (MW LD). All the strains showed the same growth performance on VW LD as on the respective standard media (SM), while ZW LD was efficient only for growth of T. obliquus and B. braunii. MW LD was the poorest growth medium for all the strains. Data on nutrient removal efficiency showed that A. maxima and T. obliquus made the best use of NH4+-N with removal values ranging between 98.9-99.8%, while P. tricornutum and B. braunii showed values of 79.0 and 88.5% respectively. Applying repeated batch cultivation in photobioreactors, the biochemical composition of A. maxima and T. obliquus biomass grown on ZW LD and VW LD, showed an increase of lipid, carbohydrates and ash in both microalgae. Biomass biochemical profiles suggest possible applications in feed, chemicals and energy sectors.

Functional ingredients from microalgae.

A wide variety of natural sources are under investigation to evaluate their possible use for new functional ingredient formulation. Some records attested the traditional and ancient use of wild harvested microalgae as human food but their cultivation for different purposes started about 40 years ago. The most popular species are Arthrospira (traditional name, Spirulina), Chlorella spp., Dunaliella spp. and Haematococcus spp. Microalgae provide a bewildering array of opportunities to develop healthier food products using innovative approaches and a number of different strategies. Compared to other natural sources of bioactive ingredients, microalgae have many advantages such as their huge biodiversity, the possibility to grow in arid land and with limited fresh water consumption and the flexibility of their metabolism, which could be adapted to produce specific molecules. All these factors led to very sustainable production making microalgae eligible as one of the most promising foods for the future, particularly as source of proteins, lipids and phytochemicals. In this work, a revision of the knowledge about the use of microalgae as food and as a source of functional ingredients has been performed. The most interesting results in the field are presented and commented upon, focusing on the different species of microalgae and the activity of the nutritionally relevant compounds. A summary of the health effects obtained together with pros and cons in the adoption of this natural source as functional food ingredients is also proposed.

Toxic effects of ZnO nanoparticles towards marine algae Dunaliella tertiolecta.

Dose response curve and population growth rate alterations of marine Chlorophyte Dunaliella tertiolecta derived from the exposure to ZnO nanoparticles were evaluated. Bulk ZnO and ionic zinc were also investigated for comparison. At the same time, the aggregation state and particle size distribution were monitored. The evaluated 50% effect concentration (EC50 1.94 [0.78-2.31] mg Zn L(-1)) indicates that nano ZnO is more toxic than its bulk counterpart (EC50 3.57 [2.77-4.80] mg Zn L(-1)). Cross-referencing the toxicity parameters calculated for ZnCl(2) (EC50 0.65 [0.36-0.70] mg Zn L(-1)) and the dissolution properties of the ZnO, it can be gathered that the higher toxicity of nano ZnO is most likely related to the peculiar physicochemical properties of the nanostate with respect to the bulk material. Furthermore growth rate of D. tertiolecta was significantly affected by nano ZnO exposure. Our findings suggest that the primary particle size of the dispersed particles affect the overall toxicity.

Embryotoxicity and spermiotoxicity of nanosized ZnO for Mediterranean sea urchin Paracentrotus lividus.

The effect of nano ZnO (nZnO) upon the fertilization and early development of embryos of the Mediterranean sea urchin Paracentrotus lividus is reported herein for the first time. Zn ion (ZnCl2) and bulk ZnO (bZnO) toxicity were assessed for comparison. The embryotoxicity tests showed a 100% effect already at 1 µM of nZnO (expressed as [Zn]) while bZnO and ZnCl2 showed EC50s of 0.98 [0.88-1.19] µM [Zn] and 2.02 [1.97-2.09] µM [Zn], respectively. Noteworthy, the frequency of developmental defects for the three compounds was dissimilar and a specific trend for larval skeletal abnormality produced by nZnO was observed. The sperm fertilization capability was only slightly affected by the tested chemicals while the effects were dramatic on the offspring quality of sperms exposed to ZnO compounds resulting in an early block of the regular larval development. ZnO toxicity seems related not only to Zinc ions but also to some surface interactions of particle/aggregates with target organisms and/or with the seawater.

Microalgae as human food: chemical and nutritional characteristics of the thermo-acidophilic microalga Galdieria sulphuraria.

The use of microalgae as a food source is still poorly developed because of the technical difficulties related to their cultivation and the limited knowledge about their chemical composition and nutritional value. The unicellular red microalga Galdieria sulphuraria has a very high daily productivity and its cultivation under acidic conditions avoided any bacterial contamination. G. sulphuraria can be cultured under autotrophic and heterotrophic conditions: in this study a screening of 43 strains showed that in the latter case a duplication of biomass production was obtained. The proximate composition (protein, carbohydrates, fiber and lipids), the micronutrient content (carotenoids, phycobiliproteins, chlorophylls and vitamins) together with the antioxidant activity of the biomass produced by a selected strain of G. sulphuraria under both cultivation conditions were determined. Results showed that the material is rich in proteins (26-32%) and polysaccharides (63-69%) and poor in lipids. Under heterotrophic cultivation conditions, the lipid moiety mainly contained monounsaturated fatty acids. Among micronutrients, some B group vitamins are present, beta-carotene is the main carotenoid and phycobiliproteins are present under both cultivating conditions. G. sulphuraria proteins are strictly associated with polysaccharide components and therefore not digestible. In the second part of the work, an extraction protocol using Viscozyme L, a commercial enzymatic preparation containing a mixture of polysaccharidases, was developed which made G. sulphuraria proteins a good substrate for human gastrointestinal enzymes. All in all, the data suggested that G. sulphuraria biomass has a potential use as food ingredients both for protein-rich or insoluble dietary fibre-rich applications. The low concentration of lipids and the absence of green color make this microalgae source particularly useful for the addition to many food preparations.

Biological activities of dermatological interest by the water extract of the microalga Botryococcus braunii.

The use of microalgae in the skin care market is already established although the scientific rationale for their benefit was not clearly defined. In this work, the biological activities of dermatologic interest of the water extract from the microalga Botryococcus braunii (BBWE) were evaluated by a battery of in vitro assays. At concentrations ranging from 0.1 to 0.001 % (w/v) BBWE promoted adipocytes differentiation by inhibiting hormone-sensitive lipase, thus promoting triglyceride accumulation in the cells. BBWE also induced gene expression of proteins involved in the maintenance of skin cells water balance such as aquaporin-3 (AQP3), filaggrin (FLG) and involucrin (INV). 0.1 % BBWE increased the gene expression of AQP3 of 2.6-folds, that of FLG and INV of 1.5- and 1.9-folds, respectively. Moreover, it induced the biosynthesis of collagen I and collagen III by 80 and 40 %, respectively, compared to the untreated control. BBWE antioxidant activity, evaluated by oxygen radical absorbance capacity (ORAC) assay, was of 43.5 µmol Trolox per gram of extract: a quite high value among those found for other microalgae extracts. BBWE inhibited the inducible nitric oxide synthase (iNOS) gene expression and the consequent nitrite oxide (NO) production under oxidative stress. At a concentration of 0.02 % BBWE reduced by 50 % the expression of iNOS and by about 75 % the NO production. Taken together, the results demonstrated that B. braunii water extract exerted an array of biological activities concurring with the skin health maintenance; therefore, it is a potential bioactive ingredient to be included in cosmetic products.