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Vibrio splendidus is a marine bacterium often considered as a threat in aquaculture hatcheries where it is responsible for mass mortality events, notably of bivalves' larvae. This bacterium is highly adapted to dynamic salty ecosystems where it has become an opportunistic and resistant species. To characterize their membranes as a first and necessary step toward studying bacterial interactions with diverse molecules, we established a labelling protocol for in vivoH solid-state nuclear magnetic resonance (SS-NMR) analysis of V. splendidus. H SS-NMR is a useful tool to study the organization and dynamics of phospholipids at the molecular level, and its application to intact bacteria is further advantageous as it allows probing acyl chains in their natural environment and study membrane interactions. In this study, we showed that V. splendidus can be labelled using deuterated palmitic acid, and demonstrated the importance of surfactant choice in the labelling protocol. Moreover, we assessed the impact of lipid deuteration on the general fitness of the bacteria, as well as the saturated-to-unsaturated fatty acid chains ratio and its impact on the membrane properties. We further characterize the evolution of V. splendidus membrane fluidity during different growth stages and relate it to fatty acid chain composition. Our results show larger membrane fluidity during the stationary growth phase compared to the exponential growth phase under labelling conditions - an information to take into account for future in vivo SS-NMR studies. Our lipid deuteration protocol optimized for V. splendidus is likely applicable other microorganisms for in vivo NMR studies.
This article was published in the following journal.
Name: Biochimica et biophysica acta. Biomembranes
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A species of gram-negative, halophilic bacteria, in the genus VIBRIO. It is considered part of normal marine flora and commonly associated with ear infections and superficial wounds exposed to contaminated water sources.
A strain of the VIBRIO CHOLERAE bacteria belonging to serogroup non-O1, infecting humans and other PRIMATES. It is related to VIBRIO CHOLERAE O1, but causes a disease less severe than CHOLERA. Eating raw shellfish contaminated with the bacteria results in GASTROENTERITIS.
A species of bacteria found in the marine environment, sea foods, and the feces of patients with acute enteritis.
A species of gram-negative bacteria in the genus VIBRIO, isolated from SHELLFISH, as well as from human diarrheal stools and ear infections.
A species of gram-negative bacteria in the genus VIBRIO, the causative agent of cold water vibriosis, a HEMORRHAGIC SEPTICEMIA of ATLANTIC SALMON.