The recent development of biological sensors has extended marine plankton studies from conducting laboratory bench work to in vivo and real-time observations. Flow cytometry (FCM) has shed new light on marine microorganisms since the 1980s through its single-cell approach and robust detection of the smallest cells. FCM records valuable optical properties of light scattering and fluorescence from cells passing in a single file in front of a narrow-collimated light source, recording tens of thousands of cells within a few minutes. Depending on the instrument settings, the sampling strategy, and the automation level, it resolves the spatial and temporal distribution of microbial marine prokaryotes and eukaryotes. Cells are usually classified and grouped on cytograms by experts and are still lacking standards, reducing data sharing capacities. Therefore, the need to make FCM data sets FAIR (Findability, Accessibility, Interoperability, and Reusability of digital assets) is becoming critical. In this paper, we present a consensus vocabulary for the 13 most common marine microbial groups observed with FCM using blue and red-light excitation. The authors designed a common layout on two-dimensional log-transformed cytograms reinforced by a decision tree that facilitates the characterization of groups. The proposed vocabulary aims at standardising data analysis and definitions, to promote harmonisation and comparison of data between users and instruments. This represents a much-needed step towards FAIRification of flow cytometric data collected in various marine environments.
To investigate the responses of a natural microbial plankton community of coastal Mediterranean waters to warming, which are still poorly known, an in situ mesocosm experiment was carried out in Thau Lagoon during autumn 2018. Several microorganisms, including virio-, bacterio-, and phytoplankton < 10 µm in size, were monitored daily and analysed using flow cytometry for 19 consecutive days in six mesocosms. Three mesocosms (control) had the same natural water temperature as the lagoon, and the other three were warmed by + 3 °C in relation to the control temperature. The cytometric analyses revealed an unexpected community dominated by picophytoplanktonic cells, including Prochlorococcus-like and Picochlorum-like cells, which had not previously been found in Thau Lagoon. The experimental warming treatment increased the abundances of nanophytoplankton, cyanobacteria, bacteria and viruses during the experiment and triggered earlier blooms of cyanobacteria and picoeukaryotes. Only the abundance of Picochlorum-like cells was significantly reduced under warmer conditions. The growth and grazing rates of phytoplankton and bacterioplankton estimated on days 2 and 8 showed that warming enhanced the growth rates of most phytoplankton groups, while it reduced those of bacteria. Surprisingly, warming decreased grazing on phytoplankton and bacteria at the beginning of the experiment, while during the middle of the experiment it decreased the grazing on prokaryote only but increased it for eukaryotes. These results reveal that warming affected the Thau Lagoon plankton community from viruses to nanophytoplankton in fall, inducing changes in both dynamics and metabolic rates.
The abundances of 17 cytometric groups encompassing four groups of bacteria, three groups of cyanobacteria, six groups of eukaryotic picophytoplankton ( 1, suggesting that predators efficiently grazed and transferred these preys. Surprisingly, the observed variations in predator abundance across seasons did not necessarily correspond with changes in grazing pressure. The significant positive relationship found between water temperature and bacterial grazing mortality rates and the reverse trend observed for larger eukaryotic prey suggested that warmer water increases small prokaryote transfer to higher trophic levels but disadvantages larger eukaryotic prey transfer.
Ce travail de thèse focalise sur le fonctionnement du réseau microbien de la lagune de Thau à travers une approche « Réseau » ayant permis l’étude simultanée de l’ensemble des groupes microbiens (21 groupes) des virus au microzooplancton. La croissance et la mortalité des micro-organismes ont été étudiées expérimentalement lors de plusieurs saisons puis d’une étude en mésocosme. Nous avons déterminé les proportions de mortalité dont sont responsables le microzooplancton et les virus au travers de la lyse virale. Un suivi in situ puis en mésocosme dans la lagune de Thau nous a permis d'observer la réponse des composantes du réseau microbien à un forçage typiquement Méditerranéen, la crue.Il apparait que les bactéries hétérotrophes ont les plus fortes croissances allant jusqu’à 2,18 jour-1. La croissance pour divers groupes pico- et nano-phytoplanctoniques était inférieure à 1,5 jour-1. La cause majeure de mortalité des différents groupes de micro-organismes étudiés est la prédation du microzooplancton (> 90% en moyenne), tandis que la lyse virale semble être un processus sporadique et nettement moins important. Le fait que les taux de prédation observés soient très proches des taux de croissance de différents micro-organismes suggère que le réseau microbien étudié présente une forte efficacité de transfert de la biomasse vers les niveaux trophiques supérieurs.Dans ce contexte, la crue provoque à court terme une diminution de la croissance microbienne et du broutage, alors qu’elle déclenche la lyse virale chez certains groupes. Cependant, la réactivité de ce réseau permet un rétablissement rapide indiquant une forte capacité de résilience de ce système.