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Notes on the Cultivation of Two Mixotrophic Dinophysis Species and Their Ciliate Prey Mesodinium rubrum.

ABSTRACT: Kleptoplastic mixotrophic species of the genus Dinophysis are cultured by feeding with the ciliate Mesodinium rubrum, itself a kleptoplastic mixotroph, that in turn feeds on cryptophytes of the Teleaulax/Plagioselmis/Geminigera (TPG) clade. Optimal culture media for phototrophic growth of D. acuminata and D. acuta from the Galician Rías (northwest Spain) and culture media and cryptophyte prey for M. rubrum from Huelva (southwest Spain) used to feed Dinophysis, were investigated. Phototrophic growth rates and yields were maximal when D. acuminata and D. acuta were grown in ammonia-containing K(-Si) medium versus f/2(-Si) or L1(-Si) media. Dinophysis acuminata cultures were scaled up to 18 L in a photobioreactor. Large differences in cell toxin quota were observed in the same Dinophysis strains under different experimental conditions. Yields and duration of exponential growth were maximal for M. rubrum from Huelva when fed Teleaulax amphioxeia from the same region, versus T. amphioxeia from the Galician Rías or T. minuta and Plagioselmis prolonga. Limitations for mass cultivation of northern Dinophysis strains with southern M. rubrum were overcome using more favorable (1:20) Dinophysis: Mesodinium ratios. These subtleties highlight the ciliate strain-specific response to prey and its importance to mass production of M. rubrum and Dinophysis cultures.

SUBMITTER: Hernandez-Urcera J

PROVIDER: S-EPMC6316069 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:During a series of blooms of Noctiluca scintillans and Mesodinium rubrum, we applied high-throughput sequencing of the 16S rRNA gene to investigate the population dynamics of free-living (FL) and particle-attached (PA) bacteria in an attempt to evaluate the influence of protozoan bloom-induced disturbances on the structuring of these two communities. Our findings revealed that the FL and PA bacterial community compositions (BCCs) displayed distinct profiles during sequential blooms, and the PA flora responded more dynamically to these pulse perturbations. The dominant bacterial groups (e.g., Flavobacteriaceae, Rhodobacteraceae, Vibrionaceae, and SAR11 subclade I) in these two communities displayed different levels of connectivity with the bloom-causative species and environmental factors. In addition, more FL bacterial groups were associated with M. rubrum, while more PA bacterial groups were related to N. scintillans. Potential endocytic bacteria of N. scintillans, particularly Vibrionaceae and Rickettsiaceae, opportunistically thrived at the peak of the bloom, suggesting that they could be important players influencing the dynamics and biogeochemical cycling of the blooms. Overall, disparities in the substrate preferences and thermal niches of various bacterial taxa as well as the short duration of the blooms (1 to 3 days) contributed to the diverse responses of the FL and PA bacterial communities to these protozoan blooms. Our research provides insight into the responses of FL and PA bacterial communities to blooms caused by protozoa like N. scintillans and M. rubrum and highlights the ecological significance of certain keystone bacterial groups during this kind of cosmopolitan protozoan bloom. IMPORTANCE Shifts in the bacterioplankton community composition during phytoplankton blooms have been studied extensively; however, investigations on protozoan blooms are rare. This study first evaluated the impact of perturbations caused by sequential protozoan blooms of the heterotrophic dinoflagellate Noctiluca scintillans and the mixotrophic ciliate Mesodinium rubrum on the structuring of these two bacterial communities. Our findings shed light on the responses of these two bacterial communities to such cosmopolitan protozoan blooms and highlight the possible ecological significance of certain keystone bacterial groups during these blooms. This research prepares the way for more focused studies that will help in understanding the roles that bacteria play during protozoan blooms and their impact on environmental health.

Project description:The development of Dinophysis populations, producers of diarrhetic shellfish toxins, has been attributed to both abiotic (e.g., water column stratification) and biotic (prey availability) factors. An important process to consider is mixotrophy of the Dinophysis species, which is an intensive feeding of the Mesodinium species for nutrients and a benefit from kleptochloroplasts. During the feeding process, the nutritional status in the environment changes due to the preference of Mesodinium and/or Dinophysis for different nutrients, prey cell debris generated by sloppy feeding, and their degradation by micro-organisms changes. However, there is little knowledge about the role of the bacterial community during the co-occurrence of Mesodinium and Dinophysis and how they directly or indirectly interact with the mixotrophs. In this study, laboratory experiments were performed to characterize the environmental changes including those of the prey present, the bacterial communities, and the ambient dissolved nutrients during the co-occurrence of Mesodinium rubrum and Dinophysis acuminata. The results showed that, during the incubation of the ciliate prey Mesodinium with its predator Dinophysis, available dissolved nitrogen significantly shifted from nitrate to ammonium especially when the population of M. rubrum decayed. Growth phases of Dinophysis and Mesodinium greatly affected the structure and composition of the bacterial community. These changes could be mainly explained by both the changes of the nutrient status and the activity of Dinophysis cells. Dinophysis feeding activity also accelerated the decline of M. rubrum and contamination of cultures with okadaic acid, dinophysistoxin-1, and pectenotoxin-2, but their influence on the prokaryotic communities was limited to the rare taxa (<0.1%) fraction. This suggests that the interaction between D. acuminata and bacteria is species-specific and takes place intracellularly or in the phycosphere. Moreover, a majority of the dominant bacterial taxa in our cultures may also exhibit a metabolic flexibility and, thus, be unaffected taxonomically by changes within the Mesodinium-Dinophysis culture system.

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Notes on the Cultivation of Two Mixotrophic Dinophysis Species and Their Ciliate Prey Mesodinium rubrum.

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