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The potential spread of infectious diseases in response to climate change and rising sea surface temperatures in temperate regions has been a growing concern for the past several decades. Extreme heat waves in the North Atlantic and North Sea regions have been correlated with an increase in human Vibrio infections; of particular concern to human health are Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus. While these species are well-known to cause disease in humans, most environmental strains are not pathogenic. Studying not only the behavior of the pathogenic strains, but that of non-pathogenic environmental isolates, may better elucidate their ecological relationship in their native microbiome and the dispersal of these species in coastal regions. Using red fluorescent protein-tagged and gentamycin-resistant V. cholerae, V. parahaemolyticus, and V. vulnificus strains, we investigated whether increasing temperatures confer greater competitive fitness to these species when incubated within a natural North Sea water sample still containing its microbiome in a small-scale niche investigation. Increased incubation temperatures alone did not confer a competitive advantage to V. cholerae, V. parahaemolyticus, and V. vulnificus. The microbial community could limit Vibrio growth at all temperatures. To the best of our knowledge, we also demonstrate the first (albeit unintentional) genetic modification of multiple species of marine bacteria through the introduction of a genetically modified V. vulnificus strain into a natural water sample in a contained system.

Future mining of polymetallic nodules in the Clarion Clipperton Zone (Northeastern Pacific) is expected to affect all benthic ecosystems. The diversity, distribution, and environmental functions of microorganisms inhabiting abyssal sediments are barely understood. To understand consequences of deep-sea mining, experimental in vitro systems needs to be established to test hypotheses on the environmental impact of mining. For this, 40 bacterial strains, belonging to proteobacteria, actinobacteria and firmicutes were isolated from deep-sea sediments and nodules sampled at depths of ≥ 4000 m. Phenotypic characterization revealed a strong inter-species and moderate intra-species variability. Determination of metal minimum inhibitory concentrations indicated the presence of acute manganese-resistant bacteria such as Rhodococcus erythropolis (228.9 mM), Loktanella cinnabarina (57.2 mM), and Dietzia maris (14.3 mM) that might be suitable systems for testing the effects of release of microbes from nodules and their interactions with sediment particles in plumes generated during mining. Comparative genomic analysis indicated the presence of manganese efflux systems relevant for future transcriptomics or proteomics approaches with environmental samples and might serve in paving the way to develop model systems including representative organisms which are currently not cultivable. Monitoring deep-sea mining activity at abyssal depth is a challenge that has to be tackled. We proposed the use of API strips as a fast on-board methodology for bacterial monitoring as an indicator for sediment plume dispersions within the water column.

Diverse microalgae-bacteria interactions play important roles for nutrient exchange processes and marine aggregate formation leading to the cycling, mineralization, or sedimentation of organic carbon in the Oceans. The main goal of this study is to report an alternative way to assess a distinct diatom-bacteria interaction. To study this at the cellular scale, an in vitro interaction model system consisting of the diatom, Thalassiosira weissflogii, and the gamma-proteobacterium, Marinobacter adhaerens HP15, had previously been established. HP15 is able to attach to T. weissflogii cells, to induce transparent exopolymeric particle formation, and to increase marine aggregation formation. Several bacterial genes important during this interaction have been studied thus far, but genes specifically expressed in co-culture remained unknown. To identify such bacterial genes, an in vivo Expression Technology (IVET) screen was employed. For this, a promoter-trapping vector containing a fusion between a promoter-less selection marker gene and a promoterless reporter gene was constructed. Construction of a library of plasmids carrying genomic fragments upstream of the fusion and its subsequent transformation into a selection marker mutant allowed detection of 30 bacterial promoters specifically expressed during the interactions with T. weissflogii. Their sequence analyses revealed that the corresponding genes could be involved in many processes such as biochemical detection of diatom cells, bacterial attachment, metabolic exchange of nitrogen compounds, and resistance toward heavy metals. Identification of genes potentially involved in branched-chain amino acid uptake and utilization confirmed our previous results of a proteomics analysis. Since our current approach identified several additional genes to be induced in co-culture, use of IVET might be a valuable complementing strategy to proteomic or transcriptomic analysis of the diatom-bacteria crossplay. The current IVET approach demonstrated that the interaction between M. adhaerens HP15 and T. weissflogii is multifactorial and is likely to involve a complex network of physiological processes.

Mangrove forests are highly productive ecosystems but represent low nutrient environments. Nitrogen availability is one of the main factors limiting mangrove growth. Diazotrophs have been identified as key organisms that provide nitrogen to these environments. N2-fixation by such organisms was found to be higher in the mangrove roots than in surrounding rhizosphere. Moreover, previous studies showed that mangroves grew better in the presence of N2-fixers indicating a potentially mutualistic relationship. However, the molecular signals and mechanisms that govern these interactions are still poorly understood. Here we present novel insights in the interaction of a diazotroph with a mangrove species to improve our understanding of the molecular and ecophysiological relationship between these two organisms under controlled conditions. Our results showed that M. mangrovicola is a versatile organism capable of competing with other organisms to survive for long periods in mangrove soils. N2-fixation by this bacterium was up-regulated in the presence of mangrove roots, indicating a possible beneficial interaction. The increase in N2-fixation was limited to cells of the exponential growth phase suggesting that N2-fixation differs over the bacterial growth cycle. Bacterial transformants harboring a transcriptional nifH::gusA fusion showed that M. mangrovicola successfully colonized mangrove roots and simultaneously conducted N2-fixation. The colonization process was stimulated by the lack of an external carbon source suggesting a possible mutualistic relationship. M. mangrovicola represents an interesting genetically accessible diazotroph, which colonize mangrove roots and exhibit higher N2-fixation in the presence of mangrove roots. Consequently, we propose this microorganism as a tool to study molecular interactions between N2-fixers and mangrove plants and to better understand how changes in the environment could impact these important and relatively unknown interactions.