Characterisation of algicidal bacterial exometabolites against the lipid-accumulating diatom Skeletonema sp.

Microalgae are of increasing interest due to their occurrence in the environment as harmful algal blooms and as a source of biomass for the production of fine and bulk chemicals. A method for the low cost disruption of algal biomass for environmental remediation or bioprocessing is desirable. Naturally-occurring algal lytic agents from bacteria could provide a cost-effective and environmentally desirable solution. A screen for algal lytic agents against a range of marine microalgae has identified two strains of algicidal bacteria isolated from the coastal region of the Western English Channel. Both strains (designated EC-1 and EC-2) showed significant algicidal activity against Skeletonema sp. and were identified as members of Alteromonas sp. and Maribacter sp. respectively. Characterisation of the two bioactivities revealed that they are small extracellular metabolites displaying thermal and acid stability. Purification of the EC-1 activity to homogeneity and initial structural analysis has identified it as a putative peptide with a mass of 1266. amu.

Characterisation of algicidal bacterial exometabolites against the lipid-accumulating diatom Skeletonema sp.

Microalgae are of increasing interest due to their occurrence in the environment as harmful algal blooms and as a source of biomass for the production of fine and bulk chemicals. A method for the low cost disruption of algal biomass for environmental remediation or bioprocessing is desirable. Naturally-occurring algal lytic agents from bacteria could provide a cost-effective and environmentally desirable solution. A screen for algal lytic agents against a range of marine microalgae has identified two strains of algicidal bacteria isolated from the coastal region of the Western English Channel. Both strains (designated EC-1 and EC-2) showed significant algicidal activity against Skeletonema sp. and were identified as members of Alteromonas sp. and Maribacter sp. respectively. Characterisation of the two bioactivities revealed that they are small extracellular metabolites displaying thermal and acid stability. Purification of the EC-1 activity to homogeneity and initial structural analysis has identified it as a putative peptide with a mass of 1266. amu.

Shirley Winifred Jeffrey 1930–2014

Australian scientist Shirley Jeffrey was a pioneer in oceanographic research, identifying the then theoretical chlorophyll c, and was a worldwide leader in the application of pigment methods in quantifying phytoplankton as the foundation of the oceanic food supply. Her research paved the way for the successful application of microalgae in aquaculture around the world. Jeffrey earned bachelor's and master's degrees at University of Sydney, majoring in microbiology and biochemistry, followed by a PhD from the King's College London Hospital Medical School. Returning to Sydney, she was hired by the Commonwealth Scientific and Industrial Research Organisation (CSIRO) to research chlorophyll c. Following this successful effort, she became a research fellow at the University of California, Berkeley from 1962 to 1964. She then became affiliated with the Scientific Committee on Oceanic Research. After a 1973 sabbatical at the Scripps Institution of Oceanography in San Diego, she returned to CSIRO, where she spent the rest of her career.

PhytoREF: a reference database of the plastidial 16S rRNA gene of photosynthetic eukaryotes with curated taxonomy

Photosynthetic eukaryotes have a critical role as the main producers in most ecosystems of the biosphere. The ongoing environmental metabarcoding revolution opens the perspective for holistic ecosystems biological studies of these organisms, in particular the unicellular microalgae that often lack distinctive morphological characters and have complex life cycles. To interpret environmental sequences, metabarcoding necessarily relies on taxonomically curated databases containing reference sequences of the targeted gene (or barcode) from identified organisms. To date, no such reference framework exists for photosynthetic eukaryotes. In this study, we built the PhytoREF database that contains 6490 plastidial 16S rDNA reference sequences that originate from a large diversity of eukaryotes representing all known major photosynthetic lineages. We compiled 3333 amplicon sequences available from public databases and 879 sequences extracted from plastidial genomes, and generated 411 novel sequences from cultured marine microalgal strains belonging to different eukaryotic lineages. A total of 1867 environmental Sanger 16S rDNA sequences were also included in the database. Stringent quality filtering and a phylogeny-based taxonomic classification were applied for each 16S rDNA sequence. The database mainly focuses on marine microalgae, but sequences from land plants (representing half of the PhytoREF sequences) and freshwater taxa were also included to broaden the applicability of PhytoREF to different aquatic and terrestrial habitats. PhytoREF, accessible via a web interface (http://phytoref.fr), is a new resource in molecular ecology to foster the discovery, assessment and monitoring of the diversity of photosynthetic eukaryotes using high-throughput sequencing.

PhytoREF: a reference database of the plastidial 16S rRNA gene of photosynthetic eukaryotes with curated taxonomy

Photosynthetic eukaryotes have a critical role as the main producers in most ecosystems of the biosphere. The ongoing environmental metabarcoding revolution opens the perspective for holistic ecosystems biological studies of these organisms, in particular the unicellular microalgae that often lack distinctive morphological characters and have complex life cycles. To interpret environmental sequences, metabarcoding necessarily relies on taxonomically curated databases containing reference sequences of the targeted gene (or barcode) from identified organisms. To date, no such reference framework exists for photosynthetic eukaryotes. In this study, we built the PhytoREF database that contains 6490 plastidial 16S rDNA reference sequences that originate from a large diversity of eukaryotes representing all known major photosynthetic lineages. We compiled 3333 amplicon sequences available from public databases and 879 sequences extracted from plastidial genomes, and generated 411 novel sequences from cultured marine microalgal strains belonging to different eukaryotic lineages. A total of 1867 environmental Sanger 16S rDNA sequences were also included in the database. Stringent quality filtering and a phylogeny-based taxonomic classification were applied for each 16S rDNA sequence. The database mainly focuses on marine microalgae, but sequences from land plants (representing half of the PhytoREF sequences) and freshwater taxa were also included to broaden the applicability of PhytoREF to different aquatic and terrestrial habitats. PhytoREF, accessible via a web interface (http://phytoref.fr), is a new resource in molecular ecology to foster the discovery, assessment and monitoring of the diversity of photosynthetic eukaryotes using high-throughput sequencing.

Macroalgae contribute to the diet of Patella vulgata from contrasting conditions of latitude and wave exposure in the UK

Analysis of gut contents and stable isotope composition of intertidal limpets (Patella vulgata) showed a major contribution of macroalgae to their diet, along with microalgae and invertebrates. Specimens were collected in areas with limited access to attached macroalgae, suggesting a major dietary component of drift algae. Gut contents of 480 animals from 2 moderately wave-exposed and 2 sheltered rocky shores in each of 2 regions (western Scotland, 55-56°N; and southwest England, 50°N), were analysed in 2 years (n = 30 site-1 yr-1). The abundance of microalgae, macroalgae and invertebrates within the guts was quantified using categorical abundance scales. Gut content composition was compared among regions and wave exposure conditions, showing that the diet of P. vulgata changes with both wave exposure and latitude. Microalgae were most abundant in limpet gut contents in animals from southwestern sites, whilst leathery/corticated macroalgae were more prevalent and abundant in limpets from sheltered and northern sites. P. vulgata appears to have a more flexible diet than previously appreciated, and these keystone grazers consume not only microalgae, but also large quantities of macroalgae and small invertebrates. To date, limpet grazing studies have focussed on their role in controlling recruitment of macroalgae by feeding on microscopic propagules and germlings. Consumption of adult algae suggests that P. vulgata may also directly control the biomass of attached macroalgae on the shore, whilst consumption of drift algae indicates that the species may play important roles in coupling subtidal and intertidal production.

Macroalgae contribute to the diet of Patella vulgata from contrasting conditions of latitude and wave exposure in the UK

Analysis of gut contents and stable isotope composition of intertidal limpets (Patella vulgata) showed a major contribution of macroalgae to their diet, along with microalgae and invertebrates. Specimens were collected in areas with limited access to attached macroalgae, suggesting a major dietary component of drift algae. Gut contents of 480 animals from 2 moderately wave-exposed and 2 sheltered rocky shores in each of 2 regions (western Scotland, 55-56°N; and southwest England, 50°N), were analysed in 2 years (n = 30 site-1 yr-1). The abundance of microalgae, macroalgae and invertebrates within the guts was quantified using categorical abundance scales. Gut content composition was compared among regions and wave exposure conditions, showing that the diet of P. vulgata changes with both wave exposure and latitude. Microalgae were most abundant in limpet gut contents in animals from southwestern sites, whilst leathery/corticated macroalgae were more prevalent and abundant in limpets from sheltered and northern sites. P. vulgata appears to have a more flexible diet than previously appreciated, and these keystone grazers consume not only microalgae, but also large quantities of macroalgae and small invertebrates. To date, limpet grazing studies have focussed on their role in controlling recruitment of macroalgae by feeding on microscopic propagules and germlings. Consumption of adult algae suggests that P. vulgata may also directly control the biomass of attached macroalgae on the shore, whilst consumption of drift algae indicates that the species may play important roles in coupling subtidal and intertidal production.

Optimisation of Spirulina platensis convective drying: evaluation of phycocyanin loss and lipid oxidation

The aim of the study was the optimisation of Spirulina platensis drying on convective hot air through the response surface methodology. The responses were thiobarbituric acid (TBA) and phycocyanin loss percentage values in final product. Experiments were carried out in perforated tray drier with parallel air flow, and the wet samples thickness and drying air temperatures were in range of 3–7 mm and 50–70 °C, respectively. The statistical analysis showed significant effect (P < 0.05) for air temperature and samples thickness. In the best drying condition, 55 °C and 3.7 mm, presented the phycocyanin loss percentage and the TBA values of approximately 37% and 1.5 mgMDA kg−1, respectively. In this drying condition, the fatty acids composition of the microalgae Spirulina did not show significance difference (P > 0.05) in relation to fresh biomass. The lipid profile of dried product presented high percentage of polyunsaturated fatty acids (34.4%), especially the gamma-linolenic acid (20.6%).

Effect of diatom supplementation during the nursery rearing of Litopenaeus vannamei (Boone, 1931) in a heterotrophic culture system

Bio-floc shrimp culture systems have been investigated in an attempt to optimize water use and prevent the discharge of effluent into the environment. The importance of microalgae in maintaining water quality and nutrition of the shrimp is well known in conventional systems; however, its maintenance amid bio-flocs and its role in the shrimp performance in this system are still poorly understood. The aim of this study was to evaluate the contribution of diatoms in the performance of Litopenaeus vannamei reared during the nursery phase in intensive system with minimal water exchange. Shrimp (0.31 ± 0.10 g) were reared among diatoms, bio-flocs and the combination of the two forming the mixture medium. The survival of shrimp was high in all treatments (90–97%). However, the shrimp reared among diatoms showed higher weight gain (P\0.05) and feed conversion ratio significantly more efficient, reaching a value of 0.47. The results indicate the importance of diatoms in bio-floc culture systems and points out to future research in an attempt to maintain a constant presence of these microalgae in culture medium without requiring successive inoculations.

Viabilidade do biodiesel de microalgas: otimização de técnicas de extração

Os problemas relacionados com o consumo energético e as emissões de poluentes relativas ao setor dos transportes representam seguramente a maior preocupação ao nível Europeu no que respeita aos gases com efeito de estufa e à poluição atmosférica. Uma das formas de resolver/minimizar estes problemas é através da aposta em combustíveis alternativos. Em particular, os biocombustíveis poderão ser uma alternativa interessante aos combustíveis convencionais. As microalgas, como matéria-prima para produção de biodiesel, apresentam-se com excelentes perspetivas de futuro e com vantagens competitivas no campo das energias renováveis. É nesta perspetiva que se enquadra o presente trabalho, cujos objetivos consistiram na extração de biodiesel a partir de microalgas secas (Nannochloropsis gaditana e Scenedesmus sp.) e na otimização das respetivas técnicas de extração lipídica. Verificou-se que, em função dos métodos e condições utilizadas, a espécie Nannochloropsis gaditana apresenta um potencial de produção de biodiesel superior à espécie Scenedesmus sp. (eficiências de extração de 24,6 (wt.%) e 9,4 (wt.%) respetivamente). Um método de rotura celular conjugado com o processo de extração lipídica via solvente orgânico é essencial, pois conseguiu-se um acréscimo de 42% no rendimento de extração, sendo 10 minutos o tempo ideal de operação. Solventes como o metanol e sistema de solventes diclorometano/metanol mostraram ser mais eficazes quando se pretende extrair lípidos de microalgas, com valores de eficiência de remoção de 30,9 (wt.%) e 23,2 (wt.%) respetivamente. De forma a valorizar os resultados obtidos no processo de extração recorreu-se à sua conversão em biodiesel através da transesterificação catalítica ácida, onde se obteve uma eficiência de conversão de 17,8 (wt.%) para a espécie Nannochloropsis gaditana.

Influence of one selected Tisochrysis lutea strain rich in lipids on Crassostrea gigas larval development and biochemical composition

Effects of a remarkably high overall lipid Tisochrysis lutea strain (T+) upon gross biochemical composition, fatty acid (FA), sterol and lipid class composition of Crassostrea gigas larvae were evaluated and compared with a normal strain of Tisochrysis lutea (T) and the diatom Chaetoceros neogracile (Cg). In a first experiment, the influence of different single diets (T, T+ and Cg) and a bispecific diet (TCg) was studied, whereas, effects of monospecific diets (T and T+) and bispecific diets (TCg and T+Cg) were evaluated in a second experiment. The strain T+ was very rich in triglycerides (TAG: 93–95% of total neutral lipids), saturated FA (45%), monounsaturated FA (31–33%) and total fatty acids (4.0–4.7 pg cell−1). Larval oyster survival and growth rate were positively correlated with 18:1n-7 and 20:1n-7, in storage lipids (SL), and negatively related to 14:0, 18:1n-9, 20:1n-9, 20:4n-6 and trans-22-dehydrocholesterol in membrane lipids (ML). Surprisingly, only the essential fatty acid 20:5n-3 in SL was correlated positively with larval survival. Correlations suggest that physiological disruption by overabundance of TAG, FFA and certain fatty acids in larvae fed T+ was largely responsible for the poor performance of these larvae. ‘High-lipid’ strains of microalgae, without regard to qualitative lipid composition, do not always improve bivalve larval performance.

Energy requirements for the continuous biohydrogen production from Spirogyra biomass in a sequential batch reactor

The current energy market requires urgent revision for the introduction of renewable, less-polluting and inexpensive energy sources. Biohydrogen (bioH2) is considered to be one of the most appropriate options for this model shift, being easily produced through the anaerobic fermentation of carbohydrate-containing biomass. Ideally, the feedstock should be low-cost, widely available and convertible into a product of interest. Microalgae are considered to possess the referred properties, being also highly valued for their capability to assimilate CO2 [1]. The microalga Spirogyra sp. is able to accumulate high concentrations of intracellular starch, a preferential carbon source for some bioH2 producing bacteria such as Clostridium butyricum [2]. In the present work, Spirogyra biomass was submitted to acid hydrolysis to degrade polymeric components and increase the biomass fermentability. Initial tests of bioH2 production in 120 mL reactors with C. butyricum yielded a maximum volumetric productivity of 141 mL H2/L.h and a H2 production yield of 3.78 mol H2/mol consumed sugars. Subsequently, a sequential batch reactor (SBR) was used for the continuous H2 production from Spirogyra hydrolysate. After 3 consecutive batches, the fermentation achieved a maximum volumetric productivity of 324 mL H2/L.h, higher than most results obtained in similar production systems [3] and a potential H2 production yield of 10.4 L H2/L hydrolysate per day. The H2 yield achieved in the SBR was 2.59 mol H2/mol, a value that is comparable to those attained with several thermophilic microorganisms [3], [4]. In the present work, a detailed energy consumption of the microalgae value-chain is presented and compared with previous results from the literature. The specific energy requirements were determined and the functional unit considered was gH2 and MJH2. It was possible to identify the process stages responsible for the highest energy consumption during bioH2 production from Spirogyra biomass for further optimisation.