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Confronting the challenge of biofilm resistance and widespread antimicrobial resistance (AMR), this study emphasizes the need for innovative monitoring methods and explores the potential of bacteriophages against bacterial biofilms. Traditional methods, like optical density (OD) measurements and confocal microscopy, crucial in studying biofilm–virus interactions, often lack real-time monitoring and early detection capabilities, especially for biofilm formation and low bacterial concentrations. Addressing these gaps, we developed a new real-time, label-free radiofrequency sensor for monitoring bacteria and biofilm growth. The sensor, an open-ended coaxial probe, offers enhanced monitoring of bacterial development stages. Tested on a biological model of bacteria and bacteriophages, our results indicate the limitations of traditional OD measurements, influenced by factors like sedimented cell fragments and biofilm formation on well walls. While confocal microscopy provides detailed 3D biofilm architecture, its real-time monitoring application is limited. Our novel approach using radio frequency measurements (300 MHz) overcomes these shortcomings. It facilitates a finer analysis of the dynamic interaction between bacterial populations and phages, detecting real-time subtle changes. This method reveals distinct phases and breakpoints in biofilm formation and virion interaction not captured by conventional techniques. This study underscores the sensor’s potential in detecting irregular viral activity and assessing the efficacy of anti-biofilm treatments, contributing significantly to the understanding of biofilm dynamics. This research is vital in developing effective monitoring tools, guiding therapeutic strategies, and combating AMR.

Organic ligands such as exopolymeric substances (EPS) are known to form complexes with iron (Fe) and modulate phytoplankton growth. However, the effect of organic ligands on bacterial and viral communities remains largely unknown. Here, we assessed how Fe associated with organic ligands influences phytoplankton, microbial, and viral abundances and their diversity in the Southern Ocean. While the particulate organic carbon (POC) was modulated by Fe chemistry and bioavailability in the Drake Passage, the abundance and diversity of microbes and viruses were not governed by Fe bioavailability. Only following amendments with bacterial EPS did bacterial abundances increase, while phenotypic alpha diversity of bacterial and viral communities decreased. The latter was accompanied by significantly enhanced POC, pointing toward the relief of C limitation or other drivers of the microbial loop. Based on the literature and our findings, we propose a conceptual framework by which EPS may affect phytoplankton, bacteria, and viruses. Given the importance of the Southern Ocean for Earth's climate as well as the prevalence of viruses and their increasingly recognized impact on marine biogeochemistry and C cycling; the role of microbe-virus interactions on primary productivity in the Southern Ocean needs urgent attention.

Organic ligands such as exopolymeric substances (EPS) are known to form complexes with iron (Fe) and modulate phytoplankton growth. However, the effect of organic ligands on bacterial and viral communities remains largely unknown. Here, we assessed how Fe associated with organic ligands influences phytoplankton, microbial, and viral abundances and their diversity in the Southern Ocean. While the particulate organic carbon (POC) was modulated by Fe chemistry and bioavailability in the Drake Passage, the abundance and diversity of microbes and viruses were not governed by Fe bioavailability. Only following amendments with bacterial EPS did bacterial abundances increase, while phenotypic alpha diversity of bacterial and viral communities decreased. The latter was accompanied by significantly enhanced POC, pointing toward the relief of C limitation or other drivers of the microbial loop. Based on the literature and our findings, we propose a conceptual framework by which EPS may affect phytoplankton, bacteria, and viruses. Given the importance of the Southern Ocean for Earth’s climate as well as the prevalence of viruses and their increasingly recognized impact on marine biogeochemistry and C cycling; the role of microbe–virus interactions on primary productivity in the Southern Ocean needs urgent attention.

recent study using Fe-limited phytoplankton strains, showed that iron (Fe) uptake rates normalized by cellular surface area were best related to dissolved iron (dFe) concentrations as the inorganic Fe (Fe’) supply rates were not sufficient to satisfy the Fe biological demand. Short-term (24 h) shipboard incubations with the in-situ phytoplankton community were used to measure Fe uptake rates that were normalized per biomass (as particulate organic carbon, POC). Fe uptake rates measured following 55FeCl3 additions (0.05 to 0.9 nM) were fitted to different Fe pools (dFe, Felabile, and Fe’) using the Michaelis-Menten equation. Data showed a similar high conditional stability constant for biological transporters across all sites and phytoplankton size classes, with only a 2-fold variation in the concentrations of cellular transporters. These observations are in line with previous reports that eukaryotic phytoplankton takes up Fe close to the limit imposed by transporters cellular density and uses similar high-affinity Fe uptake systems. To further explore the link between Fe uptake rates and Fe chemistry, we also studied the effect of Fe additions preequilibrated with different Fe-binding ligands (L) including: the siderophore desferrioxamine B, two carbohydrates (glucuronic acid and carrageenan) and two different bacterial exopolycarbohydrates (L6 and L22, referred as EPS). For all stations, phytoplankton were able to acquire Fe associated to DFB as previously reported, however, different Fe:L ratios prevent quantitative comparison with other studies. Iron bound to carbohydrates, glucuronic acid, carrageenan and EPS could enhance or decrease Fe uptake rates in comparison to equimolar FeCl3 addition. These results illustrate that the effect of such L on Fe uptake rates will depend on the in-situ plankton community and their chemical structure. The variation of the Fe’ concentrations was able to explain up to 69% of the Fe uptake rates observed for the Antarctic communit

The identification of the mechanisms by which marine dissolved organic matter (DOM) is produced and regenerated is critical to develop robust prediction of ocean carbon cycling. Polysaccharides represent one of the main constituents of marine DOM and their degradation is mainly attributed to polysaccharidases derived from bacteria. Here, we report that marine viruses can depolymerize the exopolysaccharides (EPS) excreted by their hosts using 5 bacteriophages that infect the notable EPS producer, Cobetia marina DSMZ 4741. Degradation monitorings as assessed by gel electrophoresis and size exclusion chromatography showed that 4 out of 5 phages carry structural enzymes that depolymerize purified solution of Cobetia marina EPS. The depolymerization patterns suggest that these putative polysaccharidases are constitutive, endo-acting, and functionally diverse. Viral adsorption kinetics indicate that the presence of these enzymes provides a significant advantage for phages to adsorb onto their hosts upon intense EPS production conditions. The experimental demonstration that marine phages can display polysaccharidases active on bacterial EPS lead us to question whether viruses could also contribute to the degradation of marine DOM and modify its bioavailability. Considering the prominence of phages in the ocean, such studies may unveil an important microbial process that affects the marine carbon cycle.

Exopolysaccharides (EPSs) are a class of biopolymer synthesized by Eukarya, Archea and Procarya.Bacterial EPSs are involved in biofilm establishment and biofouling phenomenon. These polymers have physicochemical and biological properties suitable with biotechnological valorization. At the opposite, their involvment in biofouling of pathogenic strains can be problematic.Enzymatic depolymerization process are necessary for EPSs structural elucidation, Bioactive oligosaccharides production or to disrupt polysaccharidic biofilms. The highlight of enzymatic phenomenon can help to understand biogeochimical process in the ocean. Nevertheless the important structural diversity as well as their complexity make the sourcing of specific enzymes difficult.Two strategies were used to find enzymes.1. The bacterial way by using EPS-producing marine strains2. The viral way, with marine bacteriophages.For the need of the study, several EPS-substrates were produced and characterized. The majority of them were totally new. An enzymatic screening on 11 marine Alteromonas strains shown that 6 were able to depolymerize their EPS in an endogenous way. A bioprospection was realized to isolates marine bacteriophages with potential viral Cazymes. 10 out of 33 phages were selectionned for their ability to be infectious with their hosts in EPS production induced. Finally, a host/virus system was chosen. The bacteriophages infecting Cobetia marina DSMZ 4741 (named Carin-1 to 5) were studied. The polysaccharidase activities of Carin-1 and Carin-5 on the L6 EPS were studied more deeply. In parallel, the complete structural elucidation of the L6 EPS was realized., Les exopolysaccharides (EPS) sont des biopolymères pouvant être synthétisés par les Eucaryotes, lesArchées et les Procaryotes. Au niveau bactérien les EPS peuvent être impliqués dans la constitution du biofilm (phénomène de biofouling) lors de la colonisation de nouveaux milieux. Ces biopolymères ont des propriétés physico-chimiques et biologiques spécifiques et innovantes à haut potentiel biotechnologique (agroalimentaire, santé, cosmétique, ingénierie environnementale ...). A l'opposé, leurs rôles écologiques lors de l'établissement de biofilms de souches potentiellement pathogènes peuvent rendre leur éradication compliquée.Les processus de dépolymérisation par voie enzymatique sont nécessaires pour réaliser l'élucidation structurale fine des EPS complexes, pour la production de dérivés bio-actifs calibrés à faible poids moléculaire ou pour empêcher la formation de biofilm. La mise en évidence de ces phénomènes enzymatiques sur des microorganismes modèles peut également permettre de mieux cerner les flux de matière au sein de certains compartiments biologiques en particulier en milieu marin. Néanmoins la complexité et grande diversité de structures des EPS rendent la recherche d’enzymes de dépolymérisation spécifiques difficile.Deux stratégies ont été employées pour trouver des sources d'enzymes.1. La voie bactérienne via l’utilisation de bactéries marines productrices d’EPS.2. La voie virale par la recherche de polysaccharidases de bactériophages marins. En plus d’EPS marins déjà connus, de nouveaux substrats (EPS) originaux ont été produits et caractérisés à partir de batéries marines d’intérêts biotechnologiques et/ou écologiques pour les besoins du projet. Un criblage enzymatique sur 11 souches bactériennes du genre Alteromonas a permis de mettre en évidence que 7 d’entre elles présentaient une activité de dépolymérisation endogène vis-à-vis de leur propre EPS. Une bioprospection a été réalisée afin de constituer une virothèque à partir d’hôtes bactériens prod