Gaël Bougaran | Université de Nantes (original) (raw)

Papers by Gaël Bougaran

Monaco/Development of Marine Resources, 2014

ABSTRACT

Biotechnology for Biofuels, 2015

Microalgae are currently emerging as one of the most promising alternative sources for the next g... more Microalgae are currently emerging as one of the most promising alternative sources for the next generation of food, feed, cosmetics and renewable energy in the form of biofuel. Microalgae constitute a diverse group of microorganisms with advantages like fast and efficient growth. In addition, they do not compete for arable land and offer very high lipid yield potential. Major challenges for the development of this resource are to select lipid-rich strains using high-throughput staining for neutral lipid content in microalgae species. For this purpose, the fluorescent dyes most commonly used to quantify lipids are Nile red and BODIPY 505/515. Their fluorescent staining for lipids offers a rapid and inexpensive analysis tool to measure neutral lipid content, avoiding time-consuming and costly gravimetric analysis. This review collates and presents recent advances in algal lipid staining and focuses on Nile red and BODIPY 505/515 staining characteristics. The available literature addresses the limitations of fluorescent dyes under certain conditions, such as spectral properties, dye concentrations, cell concentrations, temperature and incubation duration. Moreover, the overall conclusion of the present review study gives limitations on the use of fluorochrome for screening of lipid-rich microalgae species and suggests improved protocols for staining recalcitrant microalgae and recommendations for the staining quantification.

Starting microalgae continuous culture generally includes a preliminary batch culture to obtain s... more Starting microalgae continuous culture generally includes a preliminary batch culture to obtain sufficient cell density. It is possible to apply continuous regime from the beginning of the culture (IC mode) rather than to begin by an initial batch (IB mode). It is our purpose to check that both initial modes lead to identical steady-state cell characteristics. The microalga Isochrysis galbana affinis Tahiti was used for this comparative study. With an initial cell density of 5 × 10 5 cell ml -1 and a dilution rate of about 1.0 d -1 , both IB and IC modes led to identical cell density once steady-state is reached between 6 and 8 d after inoculation in the two cases. Cell concentration of chlorophyll a and pheopigment a were found to be similar for IB and IC modes at steady-state. Initial culture conditions did not influence saturation irradiance and oxygen consumption rate, which were found to be 650 ± 143 µmol phot m -2 s -1 and 1.54 × 10 -3 ± 10 -5 µmol O 2 ml -1 min -1 , respectively. At steady-state, nutrient cell uptakes were q N = 83.3 ± 2.0 fmol N cell -1 d -1 for NO 3 and q P = 5.5 ± 0.4 fmol P cell -1 d -1 for H 2 PO 4 and did not exhibit significant differences between IB and IC modes. Under the prevailing experimental conditions, results show that IC mode involved very similar steady-state cell characteristics when compared to IB mode subsequent steady-state. IC mode could be an attractive alternative especially for experimental laboratory studies, as it should lead to higher flexibility in starting continuous cultures.

ABSTRACT Gerrico visait à constituer une plate-forme d'expertise reconnue sur la gestion ... more ABSTRACT Gerrico visait à constituer une plate-forme d'expertise reconnue sur la gestion durable des ressources marines et des risques dans les espaces côtiers, tant dans le domaine des sciences de la vie et de la terre que dans celui des sciences humaines et sociales ou des sciences de l'ingénieur. Les échanges hydrologiques et sédimentaires entre le bassin versant, l'estuaire de la Loire et les eaux de la baie de Bourgneuf ainsi que les conditions de croissance et d'exposition aux risques des coquillages ont été étudiés. La perception des risques sanitaires et environnementaux par les professionnels de la baie de Bourgneuf ont fait l'objet d'une étude attentive dans le but d'explorer les moyens d'une gestion durable des ressources dont ils dépendent. De nouvelles voies d'appréhension des risques microbiologiques ont été explorées : synergie entre phycotoxines et mycotoxines, recherche de marqueurs bactériens dans certains types de contamination. De nouveaux procédés de détoxification et de sauvegarde des mollusques à visée industrielle ont été élaborés. 3 grands axes de travail ont été retenus : Axe 1 : Bioproductions et ressources marines, Axe 2 : Risques : Identification et analyse pour une gestion durable, Axe 3 : Interactions nature/société. Ce travail est le fruit d'une collaboration entre Ifremer et l'Université de Nantes.

Physiologia Plantarum, 2015

Microalgae have a diversity of industrial applications such as feed, food ingredients, depuration... more Microalgae have a diversity of industrial applications such as feed, food ingredients, depuration processes and energy. However, microalgal production costs could be substantially improved by controlling nutrient intake. Accordingly, a better understanding of microalgal nitrogen metabolism is essential. Using in silico analysis from transcriptomic data concerning the microalgae Tisochrysis lutea, four genes encoding putative high-affinity nitrate/nitrite transporters (TlNrt2) were identified. Unlike most of the land plants and microalgae, cloning of genomic sequences and their alignment with complementary DNA (cDNA) sequences did not reveal the presence of introns in all TlNrt2 genes. The deduced TlNRT2 protein sequences showed similarities to NRT2 proteins of other phyla such as land plants and green algae. However, some interesting specificities only known among Haptophyta were also revealed, especially an additional sequence of 100 amino acids forming an atypical extracellular loop located between transmembrane domains 9 and 10 and the function of which remains to be elucidated. Analyses of individual TlNrt2 gene expression with different nitrogen sources and concentrations were performed. TlNrt2.1 and TlNrt2.3 were strongly induced by low NO3 (-) concentration and repressed by NH4 (+) substrate and were classified as inducible genes. TlNrt2.2 was characterized by a constitutive pattern whatever the substrate. Finally, TlNrt2.4 displayed an atypical response that was not reported earlier in literature. Interestingly, expression of TlNrt2.4 was rather related to internal nitrogen quota level than external nitrogen concentration. This first study on nitrogen metabolism of T. lutea opens avenues for future investigations on the function of these genes and their implication for industrial applications.

Journal of Process Control, 2014

PLoS ONE, 2014

The applied exploitation of microalgae cultures has to date almost exclusively involved the use o... more The applied exploitation of microalgae cultures has to date almost exclusively involved the use of wild type strains, deposited over decades in dedicated culture collections. Concomitantly, the concept of improving algae with selection programs for particular specific purposes is slowly emerging. Studying since a decade an economically and ecologically important haptophyte Tisochrysis lutea (Tiso), we took advantage of the availability of wild type (Tiso-Wt) and selected (Tiso-S2M2) strains to conduct a molecular variations study. This endeavour presented substantial challenges: the genome assembly was not yet available, the life cycle unknown and genetic diversity of Tiso-Wt poorly documented. This study brings the first molecular data in order to set up a selection strategy for that microalgae. Following high-throughput Illumina sequencing, transcriptomes of Tiso-Wt and Tiso-S2M2 were de novo assembled and annotated. Genetic diversity between both strains was analyzed and revealed a clear conservation, while a comparison of transcriptomes allowed identification of polymorphisms resulting from the selection program. Of 34,374 transcripts, 291 were differentially expressed and 165 contained positional polymorphisms (SNP, Indel). We focused on lipid over-accumulation of the Tiso-S2M2 strain and 8 candidate genes were identified by combining analysis of positional polymorphism, differential expression levels, selection signature and by study of putative gene function. Moreover, genetic analysis also suggests the existence of a sexual cycle and genetic recombination in Tisochrysis lutea.

Current Pharmaceutical Biotechnology, 2012

Through the photosynthetic activity, microalgae process more than 25% of annual inorganic carbon ... more Through the photosynthetic activity, microalgae process more than 25% of annual inorganic carbon dissolved in oceans into carbohydrates that ultimately, serve to feed the other levels of the trophic networks. Besides, microalgae synthesize bioactive molecules such as pigments and lipids that exhibit health properties. In addition, abiotic stresses, such as high irradiance, nutrient starvation, UV irradiation, trigger metabolic reorientations ending with the production of other bioactive compounds such as ω-3 fatty acids or carotenoids. Traditionally, these compounds are acquired through the dietary alimentation. The increasing, and often unsatisfied, demand for compounds from natural sources, combined with the decrease of the halieutic resources, forces the search for alternative resources for these bioactive components. Microalgae possess this strong potential. For instance, the diatom Odontella aurita is already commercialized as dietary complement and compete with fish oil for human nutrition. In this contribution, the microalga world is briefly presented. Then, the different types of biologically active molecules identified in microalgae are presented together with their potential use. Due to space limitation, only the biological activities of lipids and pigments are described in details. The contribution ends with a description of the possibilities to play with the environmental constrains to increase the productivity of biologically active molecules by microalgae and by a description of the progresses made in the field of alga culturing.

Biotechnology and Bioengineering, 2012

Microalgae offer a high potential for energetic lipid storage as well as high growth rates. They ... more Microalgae offer a high potential for energetic lipid storage as well as high growth rates. They are therefore considered promising candidates for biofuel production, with the selection of high lipid-producing strains a major objective in projects on the development of this technology. We developed a mutation-selection method aimed at increasing microalgae neutral lipid productivity. A two step method, based on UVc irradiation followed by flow cytometry selection, was applied to a set of strains that had an initial high lipid content and improvement was assessed by means of Nile-red fluorescence measurements. The method was first tested on Isochrysis affinis galbana (T-Iso). Following a first round of mutation-selection, the total fatty acid content had not increased significantly, being 262 ± 21 mgTFA (gC)-1 for the wild type (WT) and 269 ± 49 mgTFA (gC)-1 for the selected population (S1M1). Conversely, fatty acid distribution among the lipid classes was affected by the process, resulting in a 20% increase for the fatty acids in the neutral lipids and a 40% decrease in the phospholipids. After a second mutation-selection step (S2M2), the total fatty acid content reached 409 ± 64 mgTFA (gC)-1 with a fatty acid distribution similar to the S1M1 population. Growth rate remained unaffected by the process, resulting in a 80% increase for neutral lipid productivity.

Biotechnology and Bioengineering, 2009

This study deals with the scale of a new photobioreactor for continuous microalgal production in ... more This study deals with the scale of a new photobioreactor for continuous microalgal production in hatcheries. The combination of the state-of-art with the constraints inherent to hatcheries has turned the design into a closed, artificially illuminated and external-loop airlift configuration based on a succession of elementary modules, each one being composed of two transparent vertical interconnected columns. The liquid circulation is ensured pneumatically (air injections) with respect to a swirling motion (tangential inlets). A single module of the whole photobioreactor was built-up to scale its geometry (diameter and length) and to optimize its design (air sparger, tangential inlets). The volumetric productivities were predicted by modeling radiative transfer and growth of Isochrysis affinis galbana (clone Tahiti). The hydrodynamics of the liquid phase was modeled in terms of global flow behavior (circulation and mixing times, Péclet number) and of swirling motion decay along the column (Particle Image Velocimetry). The aeration performances were determined by overall volumetric mass transfer measurements. Continuous cultures of Isochrysis affinis galbana (clone Tahiti) were run in two geometrical configurations, generating either an axial or a swirling flow. Lastly, the definitive options of design are presented as well as a 120-L prototype, currently implemented in a French mollusk hatchery and commercialized.

Bioresource Technology, 2012

Association of microalgae culture and anaerobic digestion seems a promising technology for sustai... more Association of microalgae culture and anaerobic digestion seems a promising technology for sustainable algal biomass and biogas production. The use of digestates for sustaining the growth of microalgae reduces the costs and the environmental impacts associated with the substantial algal nutrient requirements. A natural marine algae-bacteria consortium was selected by growing on a medium containing macro nutrients (ammonia, phosphate and acetate) specific of a digestate, and was submitted to a factorial experimental design with different levels of temperature, light and pH. The microalgal consortium reached a maximum C conversion efficiency (i.e. ratio between carbon content produced and carbon supplied through light photosynthetic C conversion and acetate) of 3.6%. The presence of bacteria increased this maximum C conversion efficiency up to 6.3%. The associated bacterial community was considered beneficial to the total biomass production by recycling the carbon lost during photosynthesis and assimilating organic by-products from anaerobic digestion.