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Marie, B.

Publications and source records attributed to Marie, B..

At least 19 recordsLinked to original sources

Microdiversity drives resilience and niche partitioning in the bloom forming cyanobacteria Limnospira platensis

The intraspecific evolutionary mechanisms of adaptation in blooming cyanobacteria, particularly those occurring on small spatial and temporal scales, remain poorly understood. Here, we used the unique properties of Lake Dziani Dzaha, which hosts a persistent population of Limnospira platensis inhabiting distinct macro-habitats, to explore the spatial and temporal dynamics of microevolution. The Lake Dziani Dzaha (Mayotte), very stable at least since 2010, has experienced a seismo-volcanic crisis from May 2018 to November 2019, characterized by CO2 outgassing that led to the mixing of the initially seasonally stratified water column, as well as changes in numerous physicochemical parameters. Phylogenomic analysis of 108 strains isolated during sampling campaigns in 2010, 2014, 2022, and 2024 revealed a population that remained highly phylogenetically cohesive over time. In 2022, genomic expansion was observed, characterized by an increase in genome size due to the proliferation of insertion sequences and a subsequent loss of adaptive genes. By 2024, a rapid recovery of the 2010-2014 genetic content took place through multiple independent events of horizontal gene transfer. At the spatial scale, although no significant differences in gene content were detected, distinct subpopulations were identified based on allele frequencies. Some alleles were specifically fixed and subject to different selection pressures among the subpopulations, suggesting the existence of distinct ecotypes. Overall, this study highlights the role of microdiversity in rapid resilience of blooming cyanobacteria populations within dynamic environments and in their adaptation through niche partitioning.

microbiology↗

Integrating metagenome-scale metabolic modelling and metabolomics to identify biochemical interactions in Microcystis phycospheres

Favoured by global changes, freshwater cyanobacterial harmful blooms generate major ecological, economical and public health challenges. Microcystis, one of the most widespread cyanobacterial genera, grows within a phycosphere where specialised interactions with its microbiome occur, and are suspected to influence bloom appearance and its potential toxicity. Using a combination of metagenomic, metabolomic and metabolic modelling, we characterised the phycospheres of twelve Microcystis strains isolated from a French pond. The distribution of metabolic reactions within Microcystis was consistent with their genospecies, whereas the metabolic landscape at the community level diverged from cyanobacterial phylogeny indicating functional decoupling between cyanobacteria and their associated microbiomes. Phycosphere-associated bacteria substantially expand the metabolic repertoire of the system, while maintaining functional redundancy within and across communities. On the other hand, metabolomic profiles were largely driven by cyanobacterial metabolic outputs. Metabolic modelling, together with the identification of toxic specialised metabolites produced by specific biosynthetic gene clusters, further highlighted differences in metabolic potential among phycospheres. Together, these findings deepen the understanding of Microcystis phycosphere functioning, demonstrate the value of multi-omics systems biology approaches, and underscore the ecological relevance of interspecies and inter-phycosphere metabolic interactions as a structuring process in bloom-associated microbiomes.

systems biology↗

Higher trophic status leads to more diverse and divergent microeukaryote communities over time in urban lakes from the Greater Paris (France)

Effects of trophic status on lake microeukaryote community dynamics remain underexplored. Spanning oligotrophic to hypereutrophic conditions, peri-urban lakes located in the Greater Paris region (France) offer unique opportunities to compare these dynamics along a eutrophication gradient. Here, community composition was characterized using 18S rRNA gene metabarcoding, and analyzed in the context of environmental parameters and chlorophyll a-derived trophic status throughout an 18-month sampling period. Microeukaryote assemblages were dominated by Cryptophyceae, Spirotrichea, Chrysophyceae, and Chlorophyceae, with mixotrophic and heterotrophic taxa dominating across all lakes. Taxa richness peaked at intermediate trophic levels, consistent with the intermediate disturbance hypothesis. Beta-diversity and network analyses revealed increasing community modularity, reduced connectivity, and enhanced temporal variability with higher trophic status. In lakes reaching the hypereutrophic status, communities diverged progressively over time, suggesting the onset of regime shifts. Conversely, oligo-to mesotrophic lakes maintained more connected and stable assemblages. These findings demonstrate that eutrophication fosters more diverse and increasingly divergent microeukaryote communities, underscoring its role as a central driver of microbial community restructuring in urban freshwater systems.

microbiology↗

The Neuromuscular system of Chironomus vitellinus (Diptera: Chironomidae)

Chironomids are important laboratory model organisms used to assess toxicity in freshwater environments. Cell and tissue features are not commonly used as chironomid markers to detect toxicity, but they could be extremely helpful in identifying acute and chronic effects of pollutants. The nervous system is an excellent cellular candidate since it is reactive to toxic substances. However, a detailed description of the chironomid nervous system is required prior to considering it as a candidate for a cellular toxicity marker. The present study describes the central ganglia, nerves, axons, and the neuromuscular system of Chironomus vitellinus (Freeman, 1961) to facilitate its use as a model organism in environmental studies. We find that the structure of the C. vitellinus central nervous system is identical to that observed in other Chironomus larvae. We then focused our study on the first abdominal segment and labeled the 31 hemi-segmental muscles according to a nomenclature based on their position and orientation. We also characterized their innervation and assigned the nerves a nomenclature based on their terminals location in the muscle tissue. Finally, we investigated the neuromuscular junctions (NMJs) throughout this segment and defined four types of NMJs illustrating their great variability in size and shape. We selected a model NMJ, VEL 2, and quantified its mean bouton number and muscle size. Together with documenting a neurobiological system that could be informative to insects comparative biology, these results could help establish the Chironomus NMJ as an aquatic toxicity marker.

neuroscience↗

Unveiling Crocosphaera responses to phosphorus depletion: insights from genome analysis and functional characterization

Unicellular, nitrogen-fixing cyanobacteria (UCYN) thrive and support primary production in oligotrophic oceans, playing a significant role in the marine nitrogen cycle. Crocosphaera sp, a model for studying marine nitrogen fixation, is adapted to low phosphate (Pi) conditions. Yet, how Crocosphaera copes with Pi depletion is rather poorly understood. We present genomics analysis of Pi stress-responsive genes in this genus, encompassing six C. watsonii genomes and two strains isolated in coastal environments, C. subtropica and C. chwakensis. We identified genes involved in Pi signaling, uptake, and dissolved organic phosphorus (DOP) hydrolysis. Results showed different genetic potentials to cope with Pi scarcity between the Crocosphaera strains. Physiological monitoring of cultures of C. watsonii WH8501 exposed to Pi depletion highlighted a capacity to divide several times and survive for a few more days, albeit with a skewed C:N:P stoichiometry. Upon addition of DOP, cultures efficiently recovered to a growth rate and cell composition equivalent to those observed under favorable conditions. The concomitant transcription analysis revealed diel expression patterns of Pi-related genes and endogenous clock genes, suggesting a possible circadian regulation. Our data deepen our understanding of the growth strategies Crocosphaera employs in Pi-limited environments, offering broader insights into microbial resilience in marine ecosystems.

microbiology↗

Autophagy, the Ubiquitin proteasome system, and the MAPK pathway control the temperature dependence of synaptic growth.

There is clear evidence that Earths temperature is rising at an unprecedented rate. While consequences on ecosystems are being extensively studied, little is known about the consequences of temperature on the nervous system of ectothermic animals. Here we used the Drosophila model NMJ to assess the effect of temperature on the synaptic growth of a phasic, and a tonic, motoneuron. We find that the tonic neurons synaptic size is not affected by temperature, however we do observe a temperature-dependent synaptic growth in the phasic neuron, which might be related to the increased motility observed previously at higher temperatures. We find that the level of autophagy activity changes with temperature and that autophagy genes are responsible for the temperature dependence of synaptic growth. We present evidence that this regulation could occur through the major synaptic growth regulator and ubiquitin ligase Highwire, and a pathway involving the Mitogen-Activated Protein Kinases. We present a new function for the MAPKKK, Wallenda and the MAPK P38b in directing the additional synaptic growth that takes place between 25{degrees}C and 29{degrees}C. This illustrates that temperature has different effects on a diverse population of neurons and that distinct genetic pathways are involved in regulating temperature driven changes.

neuroscience↗

A summer in the Greater Paris: trophic status of peri-urban lakes shapes prokaryotic community structure and functional potential

With more than 12 million inhabitants, the Greater Paris offers a "natural laboratory" to explore the effects of eutrophication on freshwater lakes within a relatively restricted area. Here, a four-month time-series was carried out during summertime to monitor planktonic microbial communities of nine lakes located within a [~]70 km radius around Paris (Ile-de-France, France) of comparable morphologies, yet distinct trophic statuses (mesotrophic to hypereutrophic). The contribution of the trophic status and intra-summer variations were investigated on prokaryotic community structures (16S rRNA gene sequencing) and functions (shotgun metagenomics). Sampled lakes harbored highly distinct and diverse prokaryotic communities. Although their gene pool was quite stable and shared among lakes, taxonomical and functional changes were correlated. The trophic status appears as the main driver of both community structure and functional potential. When focusing on function involved in biogeochemical cycles, responses to phosphorus limitation (mostly polyphosphate-related processes) were highlighted between trophic statuses. Hypereutrophic lakes communities displayed the highest contrast and heterogeneity over time, suggesting a regime shift compared to lakes of lower trophic status. This study explores the influence of eutrophication on lakes microbiome ecology, by comparing for the first time the planktonic microbial communities and their functional potential of lakes of distinct trophic status in close vicinity over a summer season.

microbiology↗

Limnospira (Cyanobacteria) chemical fingerprint reveals local molecular adaptation

Limnospira can colonize a wide variety of environments (e.g., freshwater, brackish, alkaline or alkaline-saline water) and develop dominant and even permanent blooms that limit over-shadowed adjacent phototrophs diversity, especially in alkaline and saline environments. Previous phylogenomic analysis of Limnospira allowed us to distinguish two major phylogenetic clades (I and II) but failed to clearly segregate strains according to their respective habitats in terms of salinity or biogeography. In the present work, we attempt to determine whether Limnospira displays metabolic signatures specific to its different habitats, particularly brackish or alkaline-saline ecosystems, and question the impact of accessory gene repertoires on respective chemical adaptations. The study of the metabolomic diversity of 93 strains of Limnospira from the Paris Museum Collection, grown under standardized lab culture conditions, showed clearly distinct chemical fingerprints that were correlated with the respective biogeographic origins of the strains. The molecules that most distinguish the different Limnospira geographic groups are sugars, lipids, peptides, photosynthetic pigments, and antioxidant molecules. Interestingly, these molecule enrichments might represent adaptation traits to the local conditions encountered in their respective sampling environments concerning salinity, light and oxidative stress. We hypothesize that within extreme environments, such as those colonized by Limnospira, a large set of flexible genes can provide remarkable adaptation to specific local environmental conditions (e.g., salinity, light, and oxidative pressure). Thus, the occurrence within Limnospira population genomes of a specific set of flexible genes potentially involved in the production of certain metabolites may provide valuable adaptative traits that may support the bloom persistence beyond environmental condition variations. ImportanceLimnospira are ubiquitous cyanobacteria able to colonize and dominate a wide range of alkaline-saline environments around the world according to remarkable adaptative strategies. Phylogenomic analysis of Limnospira platensis allowed to distinguish two major phylogenetic clades (I and II) but failed to clearly segregate strains following their habitats in terms of salinity or biogeography. One can presume that the genes found within this variable portion of the genome of these clades could be involved in L. platensis adaptation to local environmental conditions. In the present paper, we attempt to determine whether Limnospira platensis displays metabolic signatures specific to its different habitats, particularly brackish or alkalinesaline ecosystems, and question the impact of accessory gene repertoire on respective chemical adaptation.

microbiology↗

The Wingless Planar Cell Polarity pathway is essential for optimal activity-dependent synaptic plasticity.

From fly to man, the Wingless (Wg) / Wnt signaling molecule is essential for both the stability and plasticity of the nervous system. The Drosophila neuromuscular junction (NMJ) has proven to be a useful system for deciphering the role of Wg in directing activity-dependent synaptic plasticity, which, in the motoneuron, has been shown to be dependent on both the canonical and the noncanonical calcium Wg pathways. Here we show that the noncanonical planar cell polarity (PCP) pathway is an essential component of the Wg signaling system controlling plasticity at the motoneuron synapse. We present evidence that disturbing the PCP pathway leads to a perturbation in activity-dependent synaptic plasticity (ADSP). We first show that a PCP-specific allele of dishevelled (dsh) affects the de novo synaptic structures produced during ADSP. We then show that the Rho GTPases downstream of Dsh in the PCP pathway are also involved in regulating the morphological changes that take place after repeated stimulation. Finally, we show that Jun kinase is essential for this phenomenon, whereas we found no indication of the involvement of the transcription factor complex AP1 (Jun/Fos). This work shows the involvement of the neuronal PCP signaling pathway in supporting ADSP. Because we find that AP1 mutants can perform ADSP adequately, we hypothesize that, upon Wg activation, the Rho GTPases and Jun kinase are involved locally at the synapse, in instructing cytoskeletal dynamics responsible for the appearance of the morphological changes occurring during ADSP.

neuroscience↗

Tagging of water masses with covariance of trace metals and prokaryotic taxa in the Southern Ocean

Marine microbes are strongly interrelated to trace metals in the ocean. How the availability of trace metals selects for prokaryotic taxa and the potential feedbacks of microbial processes on the trace metal distribution in the ocean remains poorly understood. We investigate here the potential reciprocal links between diverse prokaryotic taxa and iron (Fe), manganese (Mn), copper (Cu), Nickel (Ni) as well as apparent oxygen utilization (AOU) across 12 well-defined water masses in the Southern Indian Ocean (SWINGS-South West Indian Ocean GEOTRACES GS02 Section cruise). Applying Partial Least Square Regression (PLSR) analysis we show that the water masses are associated with particular latent vectors that are a combination of the spatial distribution of prokaryotic taxa, trace elements and AOU. This approach provides novel insights on the potential interactions between prokaryotic taxa and trace metals in relation to organic matter remineralization in distinct water masses of the ocean.

microbiology↗

New sensitive tools to characterize meta-metabolome response to short- and long-term cobalt exposure in dynamic river biofilm communities

Untargeted metabolomics is a non-a priori analysis of biomolecules that characterizes the metabolome variations induced by short- and long-term exposures to stressors. Even if the metabolite annotation remains lacunar due to database gaps, the global metabolomic fingerprint allows for trend analyses of dose-response curves for hundreds of cellular metabolites. The combination of untargeted metabolomic features and benchmark-dose (BMD) calculations then makes it possible to determine concentration range inducing defense responses (CRIDeR) and concentration range inducing damage responses (CRIDaR). To develop this approach in a context of time-dependent microbial community changes, mature river biofilms were exposed for 1 month to four cobalt (Co) concentrations (background concentration, 1 x 10-7, 5 x 10-7 and 1 x 10- 6 M) in an open system of artificial streams. The meta-metabolomic response of biofilms was compared against a multitude of biological parameters (including bioaccumulation, biomass, chlorophyll a content, composition and structure of prokaryotic and eukaryotic communities) monitored at set exposure times (from 1 hour to 28 days). Cobalt exposure induced extremely rapid responses of the meta-metabolome, with time range inducing defense responses (TRIDeR) of around ten seconds, and time range inducing damage responses (TRIDaR) of several hours. Even in biofilms whose structure had been altered by Co bioaccumulation (reduced biomass, chlorophyll a contents and changes in the composition and diversity of prokaryotic and eukaryotic communities), CRIDeRs with similar initiation thresholds (1.41 {+/-} 0.77 x 10-10 M Co2+ added in the exposure medium) were set up at the meta-metabolome level at every time point. In contrast, the CRIDaR initiation thresholds increased by 10 times in long-term Co exposed biofilms. The present study demonstrates that defense and damage responses of biofilm meta-metabolome exposed to Co are rapidly and sustainably impacted, even within tolerant and resistant microbial communities. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=77 SRC="FIGDIR/small/567369v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@e7a87eorg.highwire.dtl.DTLVardef@e712b7org.highwire.dtl.DTLVardef@7de128org.highwire.dtl.DTLVardef@493053_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIProkaryotic community structures were impacted after 1 h of exposure to Co C_LIO_LIBiofilm meta-metabolome was impacted after 36 s of exposure to Co C_LIO_LIBiofilm meta-metabolome response was faster than changes in biofilm communities C_LIO_LIShort- and long-term exposed biofilms have similar CRIDeR initiation thresholds C_LIO_LILong-term exposed biofilms have higher CRIDaR initiation thresholds C_LI

molecular biology↗

Multi-omics analyses from a single sample: Prior metabolite extraction does not alter the 16S rRNA-based characterization of prokaryotic community in a diversity of sample types

Massive sequencing of the 16S rRNA gene has become a standard first step to describe and compare microbial communities from various samples. Parallel analysis of high numbers of samples makes it relevant to the statistical testing of the influence of natural or experimental factors and variables. However, these descriptions fail to document changes in community or ecosystem functioning. Non-targeted metabolomics are a suitable tool to bridge this gap, yet extractions protocols are different. In this study, prokaryotic community compositions are documented by 16S rRNA sequencing after direct DNA extraction, or after metabolites extraction followed by DNA extraction. Results obtained using the V3-V4 region on non-axenic cultures of cyanobacteria, lake water column, biofilm, gut of wild and lab-reared fish, indicate that prior extraction of metabolites does not influence the obtained image of prokaryotic communities. This validates sequential extraction of metabolites followed by DNA as a way to combine 16S rRNA sequencing with metabolome characterization from a single sample. This approach has the potential to complement community structure characterization with a proxy of their functioning, without the uncertainties associated with the use of separate samples.

microbiology↗

Meta-metabolomic Responses of River Biofilms to Cobalt Exposure and Use of Dose-response Model Trends as an Indicator of Effects

Metabolites are low molecular-weight molecules produced during cellular metabolism. The global expression of the meta-metabolome (metabolomics at the community level) could thus potentially be used to characterize the exposure of an organism or a community to a specific stressor. Here, the meta-metabolomic fingerprints of mature biofilms were examined after 1, 3 and 7 days of exposure to five concentrations of cobalt (0, 1 x 10-7, 1 x 10-6, 5 x 10-6 and 1 x 10-5 M) in aquatic microcosms. The global changes in meta-metabolomic fingerprints were in good agreement with those of the other biological parameters studied (cobalt bioaccumulation, biomass, chlorophyll content). To better understand the dose-responses of the biofilm meta-metabolome, the untargeted LC-HRMS metabolomic data were further processed using the DRomics tool to build dose-response model curves and to calculate benchmark doses (BMD). These BMDs were aggregated into an empirical cumulative density function. A trend analysis of the metabolite dose-response curves suggests the presence of a concentration range inducing defense mechanisms (CRIDeM) between 4.7 x 10-7 and 2.7 x 10-6 M, and of a concentration range inducing damage mechanisms (CRIDaM) from 2.7 x 10-6 M to the highest Co concentration. The present study demonstrates that the molecular defense and damage mechanisms can be related to contaminant concentrations and represent a promising approach for environmental risk assessment of metals. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=94 SRC="FIGDIR/small/545533v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@179fd06org.highwire.dtl.DTLVardef@1d2cd10org.highwire.dtl.DTLVardef@9508a3org.highwire.dtl.DTLVardef@6a1818_HPS_FORMAT_FIGEXP M_FIG C_FIG SYNOPSISThis study focuses on the interpretation of the metabolite dose-response trends in river biofilms exposed to cobalt to identify concentration range inducing cellular mechanisms and improve the environmental risk assessment of metals.

microbiology↗

Gut microbiota and holobiont metabolome composition of the Medaka fish (Oryzias latipes) are affected by a short exposure to the cyanobacterium Microcystis aeruginosa

Blooms of toxic cyanobacteria are a common stress encountered by aquatic fauna. Evidence indicates that long-lasting blooms affect fauna-associated microbiota. Because of their multiple roles, host-associated microbes are nowadays considered relevant to ecotoxicology, yet the respective timing of microbiota versus functional changes in holobionts response needs to be clarified. The response of gut microbiota and holobionts metabolome to exposure to a dense culture of Microcystis aeruginosa was investigated as a microcosm-simulated bloom in the model fish species Oryzias latipes (medaka). Both gut microbiota and gut metabolome displayed significant composition changes after only 2 days of exposure. A dominant symbiont, member of the Firmicutes, plummeted whereas various genera of Proteobacteria and Actinobacteriota increased in relative abundance. Changes in microbiota composition occurred earlier and faster compared to metabolome composition, suggesting that the microbiota drives the holobionts response. Liver and muscle metabolome were much less affected than guts, supporting that gut and associated microbiota are in the front row upon exposure. This study highlights that even short cyanobacterial blooms, that are increasingly frequent, trigger changes in microbiota composition and holobiont metabolome. It emphasizes the relevance of multi-omics approaches to explore organisms response to an ecotoxicological stress. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=118 SRC="FIGDIR/small/499308v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@112a904org.highwire.dtl.DTLVardef@16589forg.highwire.dtl.DTLVardef@1146629org.highwire.dtl.DTLVardef@3f25e4_HPS_FORMAT_FIGEXP M_FIG C_FIG Highlights- A 2-day exposure to a simulated M. aeruginosa bloom is enough to sharply decrease the Firmicute/Proteobacteria ratio in the gut of Oryzias latipes fish. - The exposure induced changes in metabolome composition after 2 days in the gut and 4 days in the liver. - The gut bacterial microbiota response occurred faster than metabolomes; we hypothesize that changes in gut microbiota may drive the gut metabolome compositional changes.

microbiology↗

Disruption of fish gut microbiota composition and holobiont metabolome by cyanobacterial blooms

BackgroundCyanobacterial blooms are one of the most common stress encountered by metazoans living in freshwater lentic systems such as lakes and ponds. Blooms reportedly impair fish health, notably through oxygen depletion and production of bioactive compounds including cyanotoxins. However, in the times of the "microbiome revolution", it is surprising that so little is still known regarding the influence of blooms on fish microbiota. In this study, an experimental approach is used to demonstrate that blooms affect fish microbiome composition and functions, as well as the metabolome of holobionts. To this end, the model teleost Oryzias latipes is exposed to simulated Microcystis aeruginosa blooms of various intensities in a microcosm setting, and the response of bacterial gut communities is evaluated in terms of composition, metagenome-encoded functions and metabolome profiling. ResultsThe gut bacterial community of O. latipes exhibits marked responses to the presence of M. aeruginosa blooms in a dose-dependent manner. Notably, abundant gut-associated Firmicutes almost disappear, while potential opportunists increase. The holobionts gut metabolome displays major changes, while functions encoded in the metagenome of bacterial partners are more marginally affected. Bacterial communities tend to return to original composition after the end of the bloom suggesting post-bloom resilience, and remain sensitive in case of a second bloom, reflecting a highly reactive gut community. ConclusionIn the context of increasingly frequent and intense blooms worldwide, results point to the relevance of accounting for short- and long-term microbiome-related effects in fish ecology, with potential outcomes relevant to conservation biology as well as aquaculture.

microbiology↗

Cnr2 is important for Ribbon Synapse Maturation and Function in Hair Cells and Photoreceptors

The role of the cannabinoid receptor 2 (CNR2) is still poorly described in sensory epithelia. We found strong cnr2 expression in hair cells (HCs) of the inner ear and the lateral line (LL), a superficial sensory structure in fish. Next, we demonstrated that sensory synapses in HCs were severely perturbed in zebrafish larvae lacking cnr2. Appearance and distribution of presynaptic ribbons and calcium channels (Cav1.3) were profoundly altered in mutant animals. Clustering of membrane-associated guanylate kinase (MAGUK) in post-synaptic densities (PSDs) was also heavily affected, suggesting a role for cnr2 for maintaining the structure of the sensory synapse. Furthermore, vesicular trafficking in HCs was strongly perturbed suggesting a retrograde action of the endocannabinoid system (ECs) mediated via cnr2 that was modulating HC mechanotransduction. We found similar perturbations in retinal ribbon synapses. Finally, we showed that larval swimming behaviors after sound and light stimulations were significantly different in mutant animals. Thus, we propose that cnr2 is critical for the processing of sensory information in the developing larva.

neuroscience↗

Toxicity, transfer and depuration of anatoxin-a (cyanobacterial neurotoxin) in medaka fish exposed by single-dose gavage

The proliferations of cyanobacteria are increasingly prevalent in warm and nutrient-enriched waters and occur in many rivers and water bodies due especially to eutrophication. The aim of this work is to study in female medaka fish the toxicity, the transfer and the depuration of the anatoxin-a, a neurotoxin produced by benthic cyanobacterial biofilms. This work will provide answers regarding acute toxicity induced by single gavage by anatoxin-a and to the risks of exposure by ingestion of contaminated fish flesh, considering that data on these aspects remain particularly limited. The oral LD50 of a single dose of ({+/-})-anatoxin-a was determined at 11.50 {micro}g.g-1. First of all, lethal dose (100% from 20 {micro}g.g-1) provokes rapid respiratory paralysis (in 1-2 min) of the fish inducing the death by asphyxia. Noticeably, no death nor apparent neurotoxicologic effect occurred during the experimentation period for the 45 fish exposed to a single sub-acute dose of ({+/-})-anatoxin-a corresponding to the no observable adverse effect level (NOAEL = 6.67 {micro}g.g-1). Subsequently, the toxico-kinetics of the ({+/-})-anatoxin-a was observed in the guts, the livers and the muscles of female medaka fish for 10 days. In parallel, a protocol for extraction of anatoxin-a has been optimized beforehand by testing 3 different solvents on several matrices, the extraction with 75% methanol + 0.1% formic acid appearing to be the most efficient. Anatoxin-a was quantified by high-resolution qTOF mass spectrometry coupled upstream to a UHPLC chromatographic chain. The toxin could not be detected in the liver after 12 h, and in the gut and muscle after 3 days. The mean clearance rates of ({+/-})-anatoxin-a calculated after 12 h are above 58%, 100% and 90% for the guts, the livers and the muscles, respectively. Non-targeted metabolomics investigations performed on the fish liver indicates that the single sub-acute exposure by gavage induces noticeable metabolome dysregulations, including important phospholipid decreases, with an organism recovery period of above 12-24h. Overall, the medaka fish do not appear to accumulate ({+/-})-anatoxin-a and to largely recover after 24h following a single sub-acute oral liquid exposure at the NOAEL.

pharmacology and toxicology↗

Subcellular localization of microcystin in the liver and the gonad of medaka fish acutely exposed to microcystin-LR

Among the diverse toxic components produced by cyanobacteria, microcystins (MCs) are one of the most toxic and notorious cyanotoxin groups. Besides their potent hepatotoxicity, MCs have been revealed to induce potential reproductive toxicity in various animal studies. However, little is still known regarding the distribution of MCs in the reproductive organ, which could directly affect reproductive cells. In order to respond to this question, an acute study was conducted in adult medaka fish (model animal) gavaged with 10 {micro}g.g-1 body weight of pure MC-LR. The histological and immunohistochemical examinations reveal an intense distribution of MC-LR within hepatocytes along with a severe liver lesion in the toxin-treated female and male fish. Besides being accumulated in the hepatocytes, MC-LR was also found in the connective tissue of the ovary and the testis, as well as in oocytes and degenerative spermatocyte-like structures. Both liver and gonad play important roles in the reproductive process of oviparous vertebrates. This observation constitutes the first observation of the presence of MC-LR in the reproductive cell of a vertebrate model with in vivo study. Our results, which provide intracellular localization of MC-LR in the gonad, advance our understanding of the potential reproductive toxicity of MC-LR in fish.

cell biology↗