Microalgae as source of functional ingredients in new-generation foods: challenges, technological effects, biological activity, and regulatory issues.

Microalgae feasibility as food ingredients or source of nutrients and/or bioactive compounds and their health effects have been widely studied. This review aims to provide an overview of the use of microalgae biomass in food products, the technological effects of its incorporation, and their use as a source of health-promoting bioactive compounds. In addition, it presents the regulatory aspects of commercialization and consumption, and the main trends and market challenges Microalgae have stood out as sources of nutritional compounds (polysaccharides, proteins, lipids, vitamins, minerals, and dietary fiber) and biologically active compounds (asthaxanthin, ß-carotene, omega-3 fatty acids). The consumption of microalgae biomass proved to have several health effects, such as hypoglycemic activity, gastroprotective and anti-steatotic properties, improvements in neurobehavioral and cognitive dysfunction, and hypolipidemic properties. Its addition to food products can improve the nutritional value, aroma profile, and technological properties, with important alterations on the syneresis of yogurts, meltability in cheeses, overrun values and melting point in ice creams, physical properties and mechanical characteristics in crisps, and texture, cooking and color characteristics in pastas. However, more studies are needed to prove the health effects in humans, expand the market size, reduce the cost of production, and tighter constraints related to regulations.

A photobioreactor using Nannochloropsis oculata marine microalgae for removal of polycyclic aromatic hydrocarbons and sorption of metals in produced water.

The objective of the present work is to evaluate the potential of the removal of PAHs and metal sorption for the treatment of petroleum produced water using a photobioreactor system with Nannochloropsis oculata microalgae. A set of photobioreactors with different gradients of produced water concentration diluted in saline water was designed, establishing five gradients (v/v): 0, 25, 50, 75 and 100%. These concentrations were established to test the removal of PAHs. The microalgal growth was monitored daily, noting the adaptation of microalgae to the addition of produced water as a culture medium, with cell growth of 5.24 × 107 cells mL-1 from 25% (v/v), 4.09 × 107 cells mL-1 from 50% (v/v), 2.77 × 107 cells mL-1 from 75% (v/v), and 1.17 × 107 cells mL-1 from 100%. The total removal efficiency of PAHs in the produced water was 94%. Organic compounds such as naphthalene, benzo(a)pyrene, benzo(b)fluoranthene, and acenaphthylene showed higher removal percentages, between 89 and 99% efficiency in produced water. Iron and zinc were the metals detected in the water produced, and iron reduced from 1.57 ± 0.08 mg L-1 to <0.1 mg L-1 after 28 days of cultivation, whereas zinc increased by 0.23 ± 0.05 to 3.90 ± 0.46 mg L-1. The PAHs removal may have occurred in two ways, by intracellular bioaccumulation or biodegradation by oxidoreductase enzymes. 0.2 g of dry biomass with maximum extraction of oil obtained 3.07% and generation of 3.70% of protein was considered as value-added products for biodiesel and bioplastics.

Enhanced Separation Capability of Sequential Injection Chromatography for Fluorimetric Determination of Intracellular Dissolved Free Amino Acids in Marine Microalgae.

This chapter describes improvements in a sequential injection method to automate the fluorimetric determination of amino acids by pre-column derivatization with o-phthaldialdehyde in presence of 2-mercaptoethanol. Separation is achieved by reversed-phase liquid chromatography in a 50 × 4.6 mm C18 silica-based monolithic column. The method is low-priced, and the separation is performed by stepwise gradient elution using six mobile phases. The mobile phase used for the first elution step is composed of methanol/tetrahydrofuran/10 mM phosphate buffer (pH 7.2) at volumetric ratio 8:1:91. Additional elution steps use mobile phases containing methanol and 10 mM phosphate buffer at volumetric ratios of 17.5:82.5, 25:75, 35:65, 50:50, and 65:35. Nineteen chromatographic peaks are observed in a mixture of twenty amino acids. The only complete co-elution is between tryptophan and methionine. The entire cycle of amino acid derivatization, chromatographic separation, and column conditioning at the end of separation lasts for 30 min. The method is successfully applied to quantify the major intracellular dissolved free amino acids in the marine microalgae Tetraselmis gracilis, Phaeodactium tricornutum, and Synechococcus elongatus.

Algevir: An Expression System for Microalgae Based on Viral Vectors.

The use of recombinant algae for the production of valuable compounds is opening promising biotechnological applications. However, the development of efficient expression approaches is still needed to expand the exploitation of microalgae in biotechnology. Herein, the concept of using viral expression vectors in microalgae was explored for the first time. An inducible geminiviral vector leading to Rep-mediated replication of the expression cassette allowed the production of antigenic proteins at high levels. This system, called Algevir, allows the production of complex viral proteins (GP1 from Zaire ebolavirus) and bacterial toxin subunits (B subunit of the heat-labile Escherichia coli enterotoxin), which retained their antigenic activity. The highest achieved yield was 1.25 mg/g fresh biomass (6 mg/L of culture), which was attained 3 days after transformation. The Algevir system allows for a fast and efficient production of recombinant proteins, overcoming the difficulties imposed by the low yields and unstable expression patterns frequently observed in stably transformed microalgae at the nuclear level; as well as the toxicity of some target proteins.

Methane production from marine microalgae Isochrysis galbana.

Methane production from marine microalgae Isochrysis galbana was assessed before and after mechanical and chemical pretreatments. Mechanical pretreatment resulted in a 61.7% increase in soluble Chemical Oxygen Demand. Different hydrolysis conditions were evaluated by varying temperature - T, sulfuric acid concentration - AC and biomass suspension concentration (measured as particulate COD - CODp) using an experimental design. The most significant interaction occurred between AC and T and the hydrolysis condition that showed the best result in the anaerobic digestion step was the condition at 40°C with addition of 0.2% (v/v) acid for 16h (9.27LCH4/kgVS). The low methane yields were attributed to inhibitory sodium concentrations for anaerobic digestion. Eliminating inhibitory sodium in the anaerobic digestion by biomass prewashing, there was a 71.5% increase in methane yield for biomass after acid hydrolysis, demonstrating the need for pretreatment and reduction in sodium concentration in the anaerobic digestion.

The phytoplankton of Guanabara Bay, Brazil: I. historical account of its biodiversity / O fitoplâncton da Baía de Guanabara, Brasil: I. histórico da biodiversidade local

This is a historical account of the biodiversity of phytoplankton in Guanabara Bay, Brazil. It is based on 57 publications that refer to sampling carried out between 1913 and 2004. The publications included are those with direct microscopic identification. Although 80 percent of the studies focus on ecological issues that tend to mention only the most abundant species, 24 publications provide comprehensive check-lists at the species level, especially of taxa > 20 μm. The inventory of species includes, to date, 308 taxa among 199 diatoms, 90 dinoflagellates, 9 cyanobacteria, 5 euglenophyceans, 1 chlorophycean, 1 prasinophycean, 1 silicoflagellate, and 2 ebriids. The most conspicuous species were the dinoflagellate Scrippsiella trochoidea and diatoms from the Skeletonema costatum complex. The first was the theme of the very first publication in the area (Faria 1914) that reported on its bloom associated with the mass mortality of fish due to oxygen depletion; it is still often found in high abundances (10(6) cell.L-1) in more protected areas. The second was long considered in the literature as a cosmopolitan and opportunistic species, until the recent discovery of cryptic species within the genus; taxonomic re-evaluation of local populations is, therefore, needed. Besides these two species, only other 25 species stood out in terms of frequency of occurrence and widespread distribution in the Bay, some known to be implicated in harmful blooms elsewhere. The biodiversity of dinoflagellates, especially within the Gymnodiniales, and that of other unidentified flagellates (Haptophyceae, Cryptophyceae, Prasinophyceae, Raphidophyceae) is largely underestimated because of the use of fixatives that distort/destroy diagnostic characters. From the initial inventory of 124 taxa published in 1917 and the subsequent additions in species numbers, one can have a false perception that the phytoplankton biodiversity has increased throughout the years, despite the overall increase in eutrophication observed in Guanabara Bay. The reason for this may be twofold: 1) it is an artifact caused by our progressively improving technical capability to detect and identify species and 2) the possible effects of eutrophication could be better perceived when the community structure is evaluated, that is, when space-time variations in the abundances of the populations (rather than just number of species) are also taken into account.

Este é o histórico dos estudos sobre a biodiversidade do fitoplâncton da Baía da Guanabara, Brasil, com base em 57 publicações relativas a amostras coletadas no período entre 1913 e 2004. Estão aqui incluídas apenas as investigações que identificaram espécies por microscopia. Apesar de 80 por cento dos estudos serem de cunho ecológico, os quais tendem a citar apenas as espécies mais abundantes, 24 publicações incluem listas completas dos táxons identificados em nível específico, especialmente daqueles > 20 μm. Atualmente, o inventário de espécies inclui 308 táxons dentre 199 diatomáceas, 90 dinoflagelados, 9 cianobactérias, 5 euglenofíceas, 1 clorofícea, 1 prasinofícea, 1 silicoflagelado e 2 ebriideos. As duas espécies que mais se destacaram foram o dinoflagelado Scrippsiella trochoidea e diatomáceas do complexo Skeletonema costatum. A primeira foi o tema de publicação pioneira sobre o fitoplâncton da Baía (Faria 1914), que relatou sua floração associada à mortandade de peixes, e continua sendo detectada em altas concentrações (10(6) cell.L-1) em áreas mais protegidas. A segunda foi considerada por muitos anos na literatura como uma espécie cosmopolita e oportunista, até a descoberta recente de espécies crípticas dentro do gênero, indicando que as populações da Baía da Guanabara requerem revisão taxonômica. Além destas duas espécies, apenas 25 apresentaram alta freqüência de ocorrência e ampla distribuição, algumas relatadas como causadoras de florações nocivas em outros locais. A biodiversidade de dinoflagelados, especialmente dentre os Gymnodiniales, assim como a de outros grupos de flagelados ainda não identificados (Haptophyceae, Cryptophyceae, Prasinophyceae, Raphidophyceae), está subestimada devido ao uso de fixadores que destorcem ou destroem caracteres diagnósticos. Se considerarmos o inventário de espécies mais antigo, uma lista de 124 táxons publicada em 1917, e o incremento no número de espécies desde então, é possível ter a falsa impressão de que a biodiversidade do fitoplâncton da Baía de Guanabara aumentou apesar da crescente eutrofização local. Há dois motivos possíveis: 1) trata-se apenas do reflexo do aprimoramento da capacidade técnica de detectar e identificar as espécies e 2) os efeitos potenciais da eutrofização podem ser melhor percebidos através da análise da estrutura da comunidade, ou seja, quando as tendências espaciais e temporais na abundância das populações (e não apenas no número de espécies) são levadas em consideração.

The phytoplankton biodiversity of the coast of the state of São Paulo, Brazil / A biodiversidade do fitoplâncton do litoral do estado de São Paulo, Brasil

The objective of this study is to compile the inventory of nearly 100 years of research about the phytoplankton species cited for the coast of the state of São Paulo, Brazil. A state-of-the-art study on the local biodiversity has long been needed to provide a baseline for future comparisons. This type of data is scattered in old scientific journals and in the so-called "grey literature", and are in need of nomenclature updating. Twenty-six publications are considered. The earliest sampling record is from 1913 (?) and the most recent from 2002. This checklist compiled from the literature was complemented with primary data collected between August 2004 and July 2006, recent surveys of the surf-zone of 20 beaches located along the coast and of a mariculture farm at the Cocanha Beach, Caraguatatuba. The complete species list includes 572 taxa: most are diatoms (82 percent), seconded by dinoflagellates (16 percent), with a minor contribution of silicoflagellates, coccolithophorids, ebriideans and cyanobacteria. The most investigated areas were: Ubatuba, São Sebastião, Santos and Cananéia. The recent surveys have a broader spatial coverage. The objectives of the studies have changed over the years from purely taxonomic to process-oriented investigations. Therefore, the longest species lists and most first records for the area were published before the 1980's and later publications, even those in which cell counts were performed, mention only the most abundant/frequent species. Electron microscopy was used for the first time in the present surveys, and new records include 38 diatoms, 42 dinoflagellates, 1 silicoflagellate, 1 ebriidean and 2 cyanobacteria. The use and interpretation of this species list require the perception of some constraints. It is not our role to question the identification made by other researchers. On the other hand, the ability to sample, analyze and identify species has evolved over the years and some considerations...

O objetivo deste estudo é compilar o inventário de quase 100 anos de pesquisa sobre a composição de espécies do fitoplâncton do litoral do Estado de São Paulo, Brasil. O retrato atualizado da biodiversidade local é necessário para establecer um dignóstico para comparações futuras. Este tipo de informação encontra-se dispersa em antigas publicações e em material considerado como "literatura cinza" e requer atualização nomenclatural. Vinte e seis publicações foram consideradas neste estudo. O registro mais antigo é de 1913 (?) e o mais recente de 2002. Esta lista de espécies, compilada a partir de dados pretéritos, foi complementada com dados primários coletados entre agosto de 2004 e julho de 2006 nas zonas de arrebentação de 20 praias paulistas e em área de maricultura da praia da Cocanha em Caraguatatuba. A lista de espécies completa inclui 572 táxons: a maioria de diatomáceas (82 por cento), seguidas por dinoflagelados (16 por cento), com uma pequena contribuição de silicoflagelados, cocolitoforídeos, ebriideas e cianobactérias. As áreas mais estudadas foram: Ubatuba, São Sebastião, Santos e Cananéia. O levantamento atual tem uma cobertura espacial mais abrangente. Ao longo dos anos, os objetivos dos estudos pretéritos variaram desde puramente taxonômicos até investigações de aspectos funcionais do ecossistema. Desta forma, os inventários mais longos e a maioria dos primeiros registros foram publicados antes de 1980 e as publicações mais recentes fazem menção apenas a espécies mais abundantes/freqüentes. Microscopia eletrônica foi empregada pela primeira vez no levantamento atual e novos registros incluem 38 diatomáceas, 42 dinoflagelados, 1 silicoflagelado, 1 ebriidea e 2 cianobactérias. O uso e interpretação deste inventário exige cautela. Se por um lado, não é viável questionar a identificação feita por outros pesquisadores, por outro lado, a capacidade de amostrar, analisar e identificar espécies evoluiu ao longo do tempo. Algumas...