Project description:Unicellular algae, termed phytoplankton, greatly impact the marine environment by serving as the basis of marine food webs and by playing central roles in biogeochemical cycling of elements. The interactions between phytoplankton and heterotrophic bacteria affect the fitness of both partners. It is becoming increasingly understood that metabolic exchange determines the nature of such interactions, but the underlying molecular mechanisms remain underexplored. Here, we investigated the molecular and metabolic basis for the bacterial lifestyle switch, from coexistence to pathogenicity, in Sulfitobacter D7 during interactions with Emiliania huxleyi, a cosmopolitan bloom-forming phytoplankter. The interaction displays two distinct phases: first, there is a coexisting phase in which the alga grows exponentially and the bacterium grows as well. The interaction shifts to pathogenic when the virulence of Sulfitobacter D7 towards E. huxleyi is invoked upon exposure to high concentrations of algal dimethylsulfoniopropionate (DMSP), which occurs when the algae reach stationary growth or when DMSP is applied exogenously to algae in exponential growth. We aimed to unravel the response of Sulfitobacter D7 to the pathogenicity-inducing compound, DMSP, and to different algae-derived infochemicals that affect the lifestyle of the bacterium. We grew Sulfitobacter D7 in conditioned media (CM) derived from algal cultures at the different growth phases, exponential and stationary (Exp-CM and Stat-CM, respectively), in which DMSP concentration is low and high, respectively. This enabled us to separate between different phases of the interaction with E. huxleyi, i.e., Exp-CM representing the coexisting phase, and Stat-CM representing the pathogenic phase. An additional pathogenicity-inducing treatment was Exp-CM supplemented with 100 µM DMSP (herein Exp-CM+DMSP). This condition mimicked co-cultures to which we added DMSP exogenously and thus induced Sulfitobacter D7 pathogenicity, which lead to death of exponentially growing E. huxleyi. In order to identify bacterial genes that are specifically responsive to DMSP, and are not affected by other algae-derived factors, we grew Sulfitobacter D7 in defined minimal medium (MM), lacking algal metabolites, supplemented with 100 µM DMSP (herein MM+DMSP), and examined the transcriptional response. After 24 h of Sulfitobacter D7 growth in all 5 media, triplicates were taken for transcriptomic analysis. Altogether, this experimental design allowed to expand our understanding on the bacterial response to DMSP, algal infochemicals and which of these are essential for coexistence and pathogenicity.
Project description:Non-starch soluble polysaccharides (NSPs) produced by yeasts are used in animal nutrition to improve health and performance. However, the magnitude of the effect may be dependent upon the quantity and the composition of the polysaccharides. As seaweeds are attractive sources of NSPs, this study was set up to evaluate their potential to improve intestinal health. The effect of NSP extracts prepared from Saccharomyces cerevisiae containing β-glucan and mannan (PSY1, positive control) or a mixture of mannanoligosaccharides (PSY2, positive control), micro algae containing β-glucan (PSA1), brown macro algae containing fucoidan and laminarin (PSA2), and green macro algae containing ulvan (PSA3) on intestinal porcine epithelial cells J2 (IPEC-J2) was studied in the presence and absence of the enterotoxigenic bacterium Escherichia coli k99 strain (ETEC) as an in vitro challenge. The E.coli-k99 strain with adhesion factor F41 (41/32) was isolated from a mastitis-infected udder. In addition, a mixed extract prepared from vegatal orgin supplemented with phenolic compounds from vegetal origin, zinc and selenium (9631), and ZnO were tested to compare responses to NSP extracts. Gene expression was measured in IPEC-J2 cells after 2 and 6 hours of incubation using “whole genome” porcine microarrays (submission as a conference paper at the SEAGRICULTURE 2017 6th International Seaweed Conference).
Project description:Micro algae's are used as alternative protein source in human and animal diets. Besides micro algae contain substantial amounts of proteins they also contain a high concentration of, often unique, biological and chemical substances with potential to induce beneficial and health promoting effects in humans and animals. This study was set up to evaluate the potential of these substances to improve (intestinal) health. The effect of extracts prepared from 3 monocultures of micro algae's (Chlorella vulgaris [C], Haematococcus pluvialis [H], and Spirulina platensis [S]) and a mixed culture of micro algae's (AM; a mixture of Scenedesmus sp. and Chlorella sp. ) was studied in the presence and absence of the enterotoxigenic bacterium Escherichia coli k99 strain (ETEC, [E]) as an in vitro challenge. The E.coli-k99 strain with adhesion factor F41 (41/32) was isolated from a mastitis-infected udder. Gene expression was measured in cultured intestinal porcine epithelium cells (IPECJ2 cell line) after 2 and 6 hours incubation with C, H, and S extracts, and after 6 hours with the AM extract, using “whole genome” porcine microarrays. Gene expression profiles were analysed using functional bioinformatics programs to provide insight in the biological processes induced by micro algae extracts.
Project description:Brown algae (Phaeophyceae) are complex photosynthetic organisms with a very different evolutionary history to green plants, to which they are only distantly related. These seaweeds are the dominant species in rocky coastal ecosystems and they exhibit many interesting adaptations to these, often harsh, environments. The brown algae are also important because they are one of only a very small number of eukaryotic lineages that have evolved complex multicellularity. This work used whole genome tiling array approach to generate a comprehensive transcriptome map of the filamentous seaweed Ectocarpus siliculosus (Dillwyn) Lyngbye, a model organism for the brown algae. Keywords: high-resolution tiling array, brown algae, ectocarpus