Environmental Microbiology

Marine algal blooms play a vital role in oceanic carbon cycling, yet the ecological consequences of algal organic matter released following their collapse via viral infection are poorly understood. Recent studies have shown that viral infection dramatically alters the hosts intracellular metabolite composition, and the subsequent viral lysate selectively promotes the growth of specific prokaryotic populations. This study aimed to elucidate the effect of organic matter derived from healthy and virus-infected cells of the bloom-forming alga Heterosigma akashiwo on the growth of heterotrophic eukaryotes, specifically Labyrinthulomycetes. These marine protists are primarily saprotrophic or predatory and contribute to dissolved organic matter (DOM) decomposition and nutrient cycling. Our field monitoring in Osaka Bay over 12 months revealed that while the overall Labyrinthulomycetes community showed no clear seasonality, specific populations of the protists co-occurred with Heterosigma akashiwo. To mechanistically investigate the potential trophic linkage suggested by these field observations, a co-culture system comprising H. akashiwo, its specific virus (HaV53), and Aurantiochytrium sp. NBRC102614, used here as a model Labyrinthulomycete, was established. In the co-culture experiments, viral lysis of H. akashiwo led to a significant increase in the cell density of Aurantiochytrium sp., demonstrating that Aurantiochytrium can thrive on substrates derived from the virus-infected alga, such as viral-induced dissolved organic matter (vDOM). These findings highlight that heterotrophic Labyrinthulomycetes are one of key consumers of virus-modified organic matter, playing a pivotal role in carbon cycling following the collapse of harmful algal blooms and influencing carbon transfer in coastal microbial food webs. IMPORTANCEMarine ecosystems are tightly regulated by the interplay between microalgae, viruses, and heterotrophic eukaryotes, yet their roles within this network have long been underestimated. Accordingly, this study aimed to provide an overview of the dynamics of environmental microalgae and heterotrophic eukaryotes, namely Heterosigma species and Labyrinthulomycetes, and to elucidate the impact of virus-infected Heterosigma akashiwo on the growth and proliferation of Aurantiochytrium species within heterotrophic Labyrinthulomycetes. This study revealed the dynamics of several Labyrinthulomycetes species associated with Heterosigma populations in coastal marine environments and demonstrated that Aurantiochytrium species have the capacity to redistribute carbon, such as by utilizing vDOM released during the termination of Heterosigma blooms via viral infection, thereby repositioning Aurantiochytrium from a passive component of Heterosigma viral infection toward an active ecological agent that facilitates energy transfer and contributes to the maintenance of microalgal community dynamics. Overall, this work provides new insights into the fate of virus-infected Heterosigma in coastal marine systems mediated by heterotrophic Labyrinthulomycetes, particularly Aurantiochytrium species, thereby filling an important knowledge gap in microbial ecology.

Bacterial communities and the bacteriophages infecting them are the basis of every ecosystem, including holobionts. The various ways in which these microorganisms interact with each other in complex communities over the life of the host affects the holobiont fitness. Despite being ubiquitous and environmentally relevant, plant-associated microbial communities remain understudied, especially in the phyllosphere, mainly because of the low abundance of microbes and the complexity of the system. In this work we followed bacteria and phage community dynamics in the phyllosphere over a growing cycle of Arabidopsis thaliana, to understand the ecology and relevance of bacteriophages in complex bacterial communities. We focused on Pseudomonas, a common plant pathogen and commensal, and the phages infecting them, in three setups of increasing complexity: in vitro, controlled experiments in planta and in wild populations of A. thaliana. We found that bacterial communities are resilient to phage infection, and more dynamic than the phages infecting them over the growing season, suggesting that although ubiquitous and abundant, bacteriophages exert selective pressures on leaf bacterial communities only intermittently.

Fungal-animal mutualisms remain significantly understudied, yet they represent some of the most successful partnerships known in nature. Fungal farming ambrosia beetles cultivate a consortium of fungal partners that include obligate filamentous members and yeasts. These fungi are maintained in highly specialized insect organs, termed mycangia, and are cultivated as food along the beetle galleries elaborated within host trees. Here we identify two previously described filamentous species, Raffaelea arxii and R. fusca, and the yeast, Ambrosiozyma monospora, as well as two new filamentous fungal species, Neocosmospora affinis and Graphium ambrosium, and two novel yeasts, designated Alloascoidea xylebori and Wickerhamomyces ambrosius, from beetle gallery walls and ambrosia beetle mycangia, using a protocol that minimizes biases in recovery by removal of a commonly used ethanol wash. To meet Koch-like postulates, we further demonstrate that all seven fungal species were individually competent at colonizing aposymbiotic X. affinis mycangia, thus demonstrating each as a viable mycangial mutualist. These data highlight methodological considerations that overcome previous limitations in mycangial content characterization, resulting in the discovery of new ambrosia beetle fungal partners. We further validate the fungal-animal mutualism by demonstrating specific colonization of the mycangial organ by potential fungal partners.

Bacteriophages are not only the most ubiquitous biological entity on earth, they also display remarkable genetic diversity across and within populations. While macrodiversity has been extensively studied, the drivers of microdiversity (intraspecies genetic diversity) remain poorly understood, particularly in relation to phage lifestyle. The distinguishing ability of temperate phages to integrate themselves into the host genome has an unknown influence on the microdiversity present. This difference in microdiversity could impact the adaptability of phages to (a)biotic factors. To identify a possible association between microdiversity and lifestyle, we analysed 12 existing viromics datasets focusing on soil bacteriophages, including 41 412 viral genomes in total. We found that phages predicted to be temperate consistently exhibit significantly higher microdiversity than their virulent counterparts in eight of 12 datasets, whereas the remaining four datasets did not show a significant trend. The detected pattern holds across multiple quality thresholds and lifestyle prediction methods. These findings suggest that lysogeny may promote or preserve genetic variation within phage populations, with potential implications for phage-host coevolution and environmental adaptability.

Emergence of multi-drug resistant (MDR) pathogens is facilitated by the mobilization of resistance genes from bacteria in animal and environmental habitats, a process often mediated by plasmids. While fertilization of agricultural soils with manure is hypothesized to serve as a pathway for transferring antimicrobial resistance plasmids to soil and crop bacteria, evidence is limited. In this study, we aimed to determine whether MDR-plasmids from manure transfer in soil, leading to the formation of long-term agricultural resistance reservoirs. To this end, we introduced a known MDR plasmid to agricultural soil where barley was subsequently grown and monitored spread of the plasmid over the course of a growing season (up to 190 days). Our experimental design mimicked conventional agricultural practices at a microcosm scale. A digital droplet PCR approach indicated plasmid transfer in the rhizosphere, which was confirmed by a targeted Hi-C method (termed Hi-C+). This demonstrated transfer of the plasmid to soil bacteria 10 days after barley planting but was not observed afterwards. The new plasmid hosts could not be identified, as plasmid-associated host Hi-C reads were absent from existing databases. This implies these hosts were rare and likely unculturable members of the soil microbiome. Our findings demonstrate that plasmid transfer from manure to soil can occur under conditions reflecting those found in agricultural settings. Furthermore, rare and uncharacterized members of the soil microbiomes may participate in acquiring MDR plasmids from manure bacteria, raising important questions about their role in spreading resistance plasmids.

Bacteria of the genus Achromatium are known for their large cell sizes and intracellular calcium carbonate deposits. Achromatium inhabit freshwater, brackish, and marine sediments where they accumulate to high abundances at the oxic-anoxic interface. These bacteria alter their vertical position in the sediment along with daily fluctuations in oxygen concentrations. Yet, the mechanism behind their migration in the sediment remains unknown. In this study, we used chemotaxis assays and time-lapse microphotography to analyze the motility and chemotactic behavior of Achromatium oxaliferum. Microscopic observations revealed that rolling and gliding were the main forms of locomotion exhibited by Achromatium. In absence of any stimulant, the movement appeared to be mostly random and changes in direction frequently occurred. Chemotaxis assays showed a negative chemotaxis of Achromatium to oxygen, sulfide, and nitrate, as evidenced by the change from undirected to directed locomotion against the respective chemical gradient. For periods of more than 1 hour, Achromatium cells moved continuously towards regions of low concentration. We further investigated whether the genetic repertoire of Achromatium corresponds to our observations. Based on lab experiments and bioinformatic analyses we conclude that Achomatium motility is propelled by type IV pili guided by a plethora of chemo- and photoreceptors. We conclude that Achromatium uses negative chemo- and phototaxis to confine their distribution in aquatic sediments between opposing oxygen and sulfide gradients. This allows Achromatium to dynamically adjust its position in redox gradients, and thus is likely to have a major contribution to its success in the global colonization of diverse aquatic sediments.

Plasmids are key drivers of horizontal gene transfer, yet their dissemination is not limited to conjugation. Extracellular vesicles (EVs) can transport plasmid DNA, but the factors governing plasmid incorporation into EVs remain poorly understood. Here, we tested whether principles of conjugative plasmid transfer, including plasmid mobility type and plasmid-plasmid interactions, extend to EV-mediated export. Using a conjugative plasmid (pKJK5) and a mobilizable plasmid (RSF1010) in two Gram-negative hosts, we quantified plasmid incorporation into EVs under single- and dual-plasmid conditions. When present individually, the conjugative plasmid was preferentially incorporated into EVs, exceeding RSF1010 by 10-23-fold despite its lower intracellular abundance. Under co-residence, this pattern reversed: RSF1010 became enriched by 13-39-fold, while pKJK5 was reduced by 2-7-fold. Consequently, EV-associated plasmid cargo shifted to RSF1010 dominance, deviating strongly from the expected 10-fold higher pKJK5 cargo if a stochastic model based on intracellular abundance and single-plasmid conditions were applicable. We propose that mobilizable plasmids under coexistence exploit conjugative plasmid transfer machinery to access membrane-associated sites, increasing their likelihood of incorporation into EVs. Our findings demonstrate that plasmid-plasmid interactions reshape EV cargo and identify a previously unrecognized mechanism that may influence extracellular gene transfer potential in microbial communities.

Pathogenic aquatic oomycetes Aphanomyces astaci and Saprolegnia parasitica represent a major threat to biodiversity and aquaculture production, but their interactions with host-associated microbes remain poorly understood. From a collection of bacterial isolates (n = 336) obtained from fish and crayfish hosts, we focused on Pseudomonas spp. (n = 70) and confirmed their previously reported strong inhibitory potential against A. astaci and S. parasitica. However, our results also revealed substantial inter- and intra-species variation in antagonism. To capture this variation, we selected eight isolates belonging to different Pseudomonas species groups (P. fluorescens, P. putida, and P. syringae) and displaying contrasting levels of anti-oomycete activity for further phenotypic assays and comparative genomic analysis. Across these isolates, mycelial inhibition was markedly stronger against A. astaci than against S. parasitica, indicating species-specific differences in susceptibility. Comparative genomic analysis revealed substantial variation in biosynthetic gene cluster (BGC) repertoires among the analysed strains. Strongly inhibitory isolates carried candidate BGCs with similarity to characterised bioactive pathways, including pyoluteorin, rhizoxin, pyrrolnitrin, DAPG, and orfamide, alongside with multiple uncharacterised clusters that were either shared among inhibitory isolates or restricted to individual strains. All analysed genomes also contained clusters related to siderophore and HCN biosynthesis. However, in vitro assays showed that siderophore production was not clearly associated with inhibitory activity and that inhibition was mediated mainly by diffusible rather than volatile compounds. Altogether, our results suggest that Pseudomonas anti-oomycete activity is species- and strain-dependent and likely reflects different combinations of multiple, predominantly diffusible metabolites rather than a single conserved mechanism. In conclusion, this study provides a foundation for future work aimed at resolving mechanisms underlying microbial antagonism toward aquatic oomycete pathogens.

Parmales (Bolidophyceae), the closest relatives of diatoms, includes isolates exhibiting one of two distinct morphotypes: a silicified non-flagellated form (S-type) and a naked flagellated form (F-type). Although alternation of these two forms for a single isolate has not been formally established, previous studies hypothesized that these morphotypes represent alternating stages of a single parmalean. In this study, we investigated the global expression patterns of S- and F-type marker genes by integrating parmalean Metagenome-Assembled Genomes (MAGs) and the metatranscriptomic dataset from Tara Oceans. We detected the expression of both S- and F-type marker genes from individual environmental MAGs. This finding provides the first metatranscriptomic evidence that natural parmalean genomes possess the potential to manifest both morphotypes. Furthermore, our analysis revealed different geographical expression patterns between the two forms. The expression of F-type markers showed a broad distribution, whereas that of S-type markers was more restricted, suggesting distinct niches for the two morpho-phases. Moreover, S-type gene expression appears to require specific environmental triggers that lead to a higher population density, whereas F-type expression is rather constitutively maintained. Overall, our results support the hypothesis of a life cycle involving morphological switching and reconcile the long-standing discrepancy between the ubiquity of parmaleans in molecular surveys and the limited geographical range for the observation of silicified cells. Based on these patterns, we propose a threshold-based model in which F-type-dominated populations persist under conditions unfavorable for growth and a morphological switch to the S-type is triggered once environmental conditions exceed a critical threshold for growth.

Symbiodiniaceae dinoflagellates are the primary photosymbionts in reef ecosystems, crucial for reef productivity. Although they are widely recognized as symbionts of animals such as corals and clams, they can occupy a broad range of reef niches, including water, sediment, and macroalgae. Understanding their ecology is typically hampered by their horizontal acquisition. Combining evidence from multiple sample types collected at the same location has the potential to address this issue, but such analyses are surprisingly rare. Here, we analysed Symbiodiniaceae communities across 74 environmental and host samples in one reef flat in Okinawa, Japan. We detected ten Symbiodiniaceae genera or genus-level clades using the ITS2 marker metabarcoding, including Clade J, previously known only from Okinotori Island, Japan. Cladocopium, Symbiodinium, and Durusdinium dominated multicellular hosts (hexacorals and Tridacna). In contrast, foraminiferal hosts were dominated by Cladocopium or genus-specific Freudenthalidium, Fugacium, and Miliolidium. Symbiont communities were mostly specific to the host genera. Water samples, with higher proportions of Durusdinium and free-living Symbiodinium, were distinct from macroalgae and sediment samples. The latter did not differ significantly from each other and contained Freudenthalidium, Fugacium, Miliolidium, Clade I, and Clade J. Only three ITS2 variants were shared across all sample categories, but many variants were unique to hosts or habitats. We highlight that both unicellular and multicellular hosts harbor specific endosymbiont types, with lower diversity than in the surrounding environment. Our results imply that host diversity, availability, and environmental context jointly structure photosymbiont communities at fine spatial scales within coral reef ecosystems.

Foods are spatially structured and heterogeneous matrices in which microbial pathogens predominantly grow as immobilised microcolonies rather than planktonic free cells. However, most predictive microbiology and risk assessment models rely on homogeneous liquid cultures, potentially overlooking spatial effects on stress adaptation. Here, we investigated how growth within food-like semi-solid matrices influences stress adaptation and digestive tolerance of major foodborne pathogens. We compared planktonic and spatialised lifestyles across multiple species exposed to salt and organic acid stresses. Spatial growth profoundly altered growth dynamics in a stress- and species-dependent manner. Notably, spatial growth markedly enhanced tolerance to simulated gastrointestinal stresses in vitro, particularly under acidic conditions. This protective effect was further confirmed in vivo within the acidic midgut of Hermetia illucens larvae. Our findings demonstrate that spatial organisation generates distinct physiological states that increase pathogen resilience, highlighting the need to integrate spatialisation into predictive models and quantitative microbial risk assessment.

SARS{square}CoV{square}2 infection is associated with marked changes of the upper respiratory tract mycobiome. URT mycobiome Changes in non-hospitalized patients however, remains poorly defined. We performed shotgun metagenomic sequencing of 95 upper respiratory tract swab samples from 48 symptomatic SARS{square}CoV{square}2-positive individuals and 47 healthy controls from central India. Fungal diversity and community structure were compared using alpha- and beta-diversity analyses, while differential taxa were identified using prevalence-based testing and LEfSe. SARS{square}CoV{square}2-positive samples showed significantly higher fungal alpha diversity than controls, with increased Shannon diversity (p = 0.000319) and Simpson diversity (p = 0.017). Beta-diversity analysis showed significant separation between groups for both Bray-Curtis and Jaccard distances (PERMANOVA p = 0.001), with significant dispersion effects as well (PERMDISP p = 0.001). Differential analysis identified more SARS{square}CoV{square}2-enriched than control-enriched taxa, including Candida orthopsilosis, Malassezia furfur, M. sympodialis, M. globosa, Aspergillus niger, A. terreus, and A. nidulans. Aspergillus sydowii was the main control-enriched taxon. LEfSe and concordant multi-test analysis supported these findings, and sensitivity analysis confirmed robustness across thresholds. Certain SARS{square}CoV{square}2-enriched taxa were linked to confirmed or probable COVID{square}19-associated fungal infections, whereas no such pathogens were detected in controls. These findings indicate that SARS{square}CoV{square}2 infection is associated with URT mycobiome dysbiosis and enrichment of clinically relevant opportunistic fungi in community cases.

Bacterial adaptation to fluctuations in salinity includes the intracellular accumulation of organic compounds called compatible solutes (CS) such as the amino acid derivatives ectoine and 5-hydroxyectoine. These compounds also play a less appreciated role as readily available nutrients, scavenged from dissolved organic matter in both marine and terrestrial environments. Vibrio diabolicus is a marine bacterium originally isolated from deep-sea hydrothermal vents and later shown to have worldwide distribution. In this work, we demonstrated the biosynthesis and uptake of CS ectoine and glycine betaine under high osmotic stress conditions, but not in unstressed V. diabolicus cells. A region on chromosome 1 of V. diabolicus strain 3098 encoded homologues of genes for ectoine and 5-hydroxyectoine catabolism (eutDE, ssd_atf_eutBCA), regulation (asnC, enuR), and transport (ectoine TRAP-type uehP, uehQM). Our data showed that ectoine was used as a high energy yielding sole carbon source and the eutD gene was essential for ectoine consumption. Phylogenetics based on EutD (DoeA) and gene neighborhood analyses showed that a catabolism cluster was present in Proteobacteria, Thermosulfobacteriota, Bacillota, Actinomycetota, and Archaea. The cluster had a limited phylogenetic distribution in Gammaproteobacteria and Betaproteobacteria and was widespread in Alphaproteobacteria. Phylogenetic reconstruction was consistent with vertical inheritance with gene loss with repeated horizontal acquisitions of the pathway across lineages. The ectoine catabolism pathway was vertically inherited in Halomonadaceae and Vibrionaceae, with patterns of gene and pathway loss. Betaproteobacteria Burkholderia, Caballeronia, and Paraburkholderia EutD proteins clustered together and EutD from most Pseudomonas species shared a most recent common with this group. EutD from Alphaproteobacteria branched in eight divergent clusters with long branch lengths but showed a remarkable conservation of synteny. Catabolism and transporter genes in this group were contiguous and contained either a TRAP-type UehPQM or an ABC-type EhuABCD ectoine transporter. Gram-positive bacteria and Archaea were not previously shown to consume ectoines, however, we identified putative ectoine catabolism clusters among Bacilli, Clostridia, Actinomycetes, and Halobacteria. IMPORTANCEEctoine is a well-established CS used to overcome osmotic stress, produced by a wide range of bacteria. The demonstration of ectoine biosynthesis and catabolism in V. diabolicus showed that it is conditionally utilized as an osmoprotectant or a nutrient source depending on environmental cues. The conservation of large syntenic blocks of ectoine catabolism, transport, and regulatory genes suggested strong selective pressure to maintain this trait. EutD (DoeA) phylogeny patterns largely followed taxonomy with evidence of horizontal transfer in specific clades, and showed ectoine consumption has a broad taxonomic spread and is lineage enriched. In our dataset, Alphaproteobacteria contained the largest diversity of EutD lineages; Gammaproteobacteria from marine environments formed a strong secondary group; and EutD from Betaproteobacteria were the least diverse. Many species that contained EutD are associated with saline, marine, and plant-associated niches, where ectoine can be scavenged as a nutrient source. The identification of a putative ectoine catabolism pathway in Gram-positive bacteria and Archaea needs to be experimentally confirmed and suggests undiscovered diversity to be revealed by future genome sequencing.

Microbial habitats that receive repeated external input may not remain shaped by that input forever if local retention allows resident communities to build up over time. Here, we used a controlled bench-scale sink p-trap system to examine how community assembly unfolded during initial establishment in new, bleach-treated p-traps. Two p-traps received repeated handwashing-water input, while one received tap water as baseline. The treated p-traps, but not the control, showed clear successional change toward later resident-like states. Nested-model comparisons further showed that recent external input had its greatest influence early in succession, but the p-traps own prior state remained the stronger predictor throughout. Final-day post-flush trajectories indicated short-term displacement from pre-flush positions, with later time points tending to move back toward late-stage resident centroids. Together, these results show that repeated inoculation does not necessarily keep communities under continued outside influence. Instead, retentive microbial habitats can shift over time from early sensitivity to external input toward persistence shaped more by local history.

The pillar coral Dendrogyra cylindrus is a rare but iconic member of Caribbean reefs that has suffered range-wide losses. D. cylindrus is highly susceptible to stony coral tissue loss disease (SCTLD), and the outbreak has contributed to the functional extinction of Floridas population of pillar corals. The coral microbiome can impact the health and disease resistance of coral colonies, yet little is known about what constitutes the core microbiome of D. cylindrus. This information is crucial for comparisons of healthy and diseased tissue in pathogen identification studies and can be applied to restoration efforts as a coral health metric. Therefore, we characterized the microbiomes of D. cylindrus colonies ahead of the SCTLD disease front in Belize and Curacao. The most prevalent members of the D. cylindrus microbial community were bacteria for which taxonomy could not be assigned confidently beyond the level of domain as well as the putatively endosymbiotic genera Endozoicomonas, Ca. Amoebophilus, and Spiroplasma. The coral reefs of Belize and Curacao represent distinct Caribbean marine ecoregions, and we documented regional differences in strains among predominant bacterial taxa. The understudied microbiome of D. cylindrus harbors unique bacterial lineages that are in danger of extinction along with its critically endangered coral host, and these bacterial lineages may be important bioindicators during restoration efforts. IMPORTANCETropical corals face global extinction if average temperatures rise by 2{degrees}C (3.6{degrees}F), which may occur as soon as 2050. Included in the loss of charismatic macrofauna like the majestic pillar coral is the loss of the biological and genetic diversity of its symbionts. Here we examined the bacterial and archaeal communities associated with Caribbean pillar corals and found that the microbiome was dominated by taxonomically unclassified and putatively endosymbiotic taxa. Endosymbiotic bacteria, which live inside the coral tissue, are likely to have evolved unique adaptations to become symbionts and may be important to the health and success of pillar corals in ecosystem restoration efforts.

In response to the comparatively sudden application of industrial scale levels of antibiotics to an ecosystem where naturally produced antibiotics are scarce, namely the ecologies within and around agricultural settings, animal-associated microbes have had to rapidly adapt, mostly relying on mobile genetic elements (MGEs) taken up due to loss of CRISPR functionality. Due to selection for resistance and other adaptive traits carried by dynamic and rapidly recombining MGEs, plasmids and transposons have rapidly accumulated in this human-proximal environment. Because of the occurrence of Enterococcus faecalis in a wide range of hosts up and down the food chain, and the fact that this species represents the greatest generalist of the genus, we comprehensively examined the mobilome of multidrug-resistant E. faecalis (ST330, ST591, ST710, and ST711) from healthy piglets raised on dispersed Brazilian farms, using long-read sequencing, analysis of plasmid pangenomes, and conjugation assays with these strains serving as donors. Genomes ranged from [~]2.8-3.1 Mb, with diverse MGEs constituting [~]7-15% of those genomes. Large modular antimicrobial resistance-encoding gene blocks ([~]40 Kb) were observed to be integrated into a [~]67 Kb chromosomal segment of the pathogenicity island AF454824. Prophages contributed up to 70% of the chromosomal mobile element content, integrating into both CRISPR-deficient genomes and those with intact type II-A CRISPR1 arrays, which were enriched with Caudoviricetes phage-targeting spacers across all strains. Plasmid content showed pronounced mosaicism driven by diverse insertion sequences, transposons, and related mobile elements, many directly implicated in AMR gene cluster acquisitions. RepA_N, Inc18, and Rep3 plasmids, mostly conjugative, also carried various persistence-related traits, suggesting they may actively enhance agricultural fitness rather than passively accumulate due to loss of CRISPR protection.

The Canadian Arctic Archipelago (CAA) is warming at an unprecedented rate, leading to sea ice loss and glacial retreat. Marine-terminating (tidewater) glaciers can fuel summertime marine productivity by delivering nutrient-rich deep waters via upwelling to the surface ocean. While the impact of glacier-induced upwelling has been well-studied in the context of phytoplankton and primary productivity, its effects on broader marine microbial communities remain poorly understood. We investigated how glacier-driven upwelling shapes marine microbial (bacterial and archaeal) communities across a series of sites in the CAA. At upwelling sites, the upper 50 m of the water column exhibited elevated nutrient concentrations and physical characteristics that resembled deeper waters, which were associated with differences in microbial community composition relative to non-upwelling sites. Our results indicate that upwelling influences microbial communities in surface waters in two ways. It directly introduces typically deeper-water-associated taxa into surface waters and reshapes ecological niches by enhancing nutrient supply and stimulating primary production, indirectly driving changes in microbial communities. The enrichment of Candidatus Nitrosopumilus, a deep water nitrifier, likely affects nitrogen cycling and raises the possibility of active nitrification in surface waters. Likewise, the increased abundance of taxa known to be associated with phytoplankton-derived organic matter in upwelling regions suggests an enhanced capacity to process organic matter generated from elevated primary productivity. Ultimately, as tidewater glaciers continue to retreat, the resulting changes in the glacially-driven upwelling regime will likely shift marine microbial communities towards assemblages adapted to less productive ecosystems, with implications for nutrient cycling in these systems. ImportanceClimate change has a disproportionate impact on the Arctic, with rising temperatures causing increased marine-terminating glacier retreat and changes in the marine water column structure. The consequent loss of the ability of these glaciers to upwell deep water to the surface ocean results in a reduction of nutrient delivery and mixing in these ecosystems. Previous work has highlighted the importance of marine-terminating glaciers in sustaining phytoplankton productivity during the summer season through this delivery of deep-water nutrients to the surface ocean. The impact of glacially-induced upwelling on marine bacterial and archaeal communities, however, remains underexplored. We found that in regions with glacially-driven upwelling, the surface ocean showed enrichment of phytoplankton-associated taxa and nitrifiers commonly associated with deep waters. This work underscores the role of glacially-driven upwelling in structuring both microbial communities and nutrient cycling, suggesting that glacier loss could reshape community composition and biogeochemical processes in a rapidly changing Arctic.

Coral white syndrome (WS) is a widespread condition characterized by tissue loss and skeletal exposure, with multiple bacterial pathogens--particularly Vibrio species--implicated in its etiology. To investigate the genomic basis of potential virulence in Vibrio associated with WS of Porites cylindrica, we isolated 57 Vibrio strains from healthy and diseased coral tissues collected from reefs in Guam. Thirteen representative strains from five dominant species--V. coralliilyticus, V. owensii, V. tubiashii, V. harveyi, and V. tetraodonis--were selected for whole-genome sequencing using Oxford Nanopore Technology. Comparative genomic analyses revealed a conserved repertoire of extracellular enzymes, including hemolysins, cytolysins, metalloproteases, and subtilisin-like peptidases, alongside lineage-specific toxin and regulatory modules. Variation in secretion systems (T1SS-T6SS), particularly in T3SS, T4SS, and T6SS subtypes, reflected diversification of host-interaction and competitive capacities across species. Mobile genetic elements, including plasmids and prophages, contributed additional virulence-associated genes and secretion clusters, underscoring the role of horizontal gene transfer in shaping the accessory genome content. Notably, V. coralliilyticus harbored cholera toxin-related genes (ace and zot), while conjugative plasmid systems indicated potential for gene dissemination across lineages. Together, these findings demonstrate that virulence potential in coral-associated Vibrio is broadly distributed and structured by a conserved ecological core overlaid with flexible, horizontally acquired modules. This study provides the first comparative genomic framework for Vibrio associated with P. cylindrica, advancing our understanding of how genomic plasticity and modular virulence repertoires may contribute to opportunistic disease dynamics in coral reef ecosystems.

The plant-beneficial bacterium Pseudomonas protegens CHA0 (CHA0) is widely studied for the biological control of soil-borne plant diseases. Beyond its root-colonising capabilities, CHA0 can also infect and kill insect larvae and thus exhibits a multi-host lifestyle shared with other plant- and insect-colonising bacteria. To better understand the robustness of this multi-host lifestyle, we subjected CHA0 to ten consecutive passages through larvae of the pest insect Plutella xylostella via repeated cycles of insect colonisation and killing forcing it into an insect-only lifestyle. Overall, serial passaging did not result in consistent changes in insect killing speed, larval or root colonisation, plant protection efficiency, microbial antagonism or in vitro growth. This suggests that its multi-host lifestyle was conserved following serial passage. Nonetheless, a few independently passaged lines showed an increase in larval killing speed, which in one case might be linked to choline uptake. To disentangle changes specific to the insect host from those arising due to the experimental system itself, we conducted parallel serial passages through the same system while omitting the insect host. In some of these lines, exposure to the background of the system led to changes in microbial antagonism and in in vitro growth, which likely are associated with mutations in regions encoding for regulatory systems. Our findings indicate that P. protegens CHA0 remains phenotypically stable in complex environments such as an insect host, suggesting that the multi-host lifestyle might also be conserved when applied in the field and supporting CHA0s potential for reliable biocontrol performance against both plant diseases and insect pests. Author summaryControlling insect pests with living organisms, known as biological control, offers an environmentally friendly alternative to chemical pesticides. The plant-beneficial bacterium Pseudomonas protegens CHA0 is a promising biocontrol candidate that not only colonizes plant roots but also infects and kills certain insect larvae. This ability to colonize different hosts appears to be a conserved trait also observed in other bacteria. To better understand the robustness of this multi-host lifestyle, we repeatedly exposed CHA0 to larvae of the insect pest Plutella xylostella and assessed the resulting physiological and genetic changes. Surprisingly, after ten cycles, CHA0 largely retained its insect-killing and plant-protective traits. Although a few populations showed minor changes, including slightly faster insect killing and traits associated with aspects of the experimental system, these changes were limited in scope. Overall, our findings suggest that P. protegens CHA0 does not change rapidly in complex environments such as an insect host, supporting its potential for reliable biocontrol performance in the field.

Ostreobium, a siphonous green alga capable of living inside of calcium carbonate substrates, including the skeletons of reef-building corals. This study investigates spectral niche preferences and physiological strategies of Ostreobium using community-wide experiments. We exposed natural Ostreobium communities from Porites lutea collected across shallow, mid, and deeper-water sites to three light conditions: far-red, blue, and white light, simulating healthy shallow-water corals, deeper water conditions, and bleached coral skeletons respectively. Using 16S rRNA metabarcoding and chlorophyll analysis, we assessed community changes and physiological responses over 16 weeks. We show significant variation in spectral preferences among Ostreobium OTUs, with clear evidence for both generalist and specialist strategies. Chlorophyll analysis showed photoacclimation responses through changes in pigment compositions. Our work shows that the spectral architecture of the reef plays a role in structuring Ostreobium communities, but the many mismatches between spectral preferences of OTUs and their observed presence in nature, suggests that inter-species competition is likely to be an even stronger contributor to community structure across the reefs microhabitats. We show that physiological heterogeneity within Ostreobium is strongly phylogenetically structured, and our results clearly highlight the importance of considering OTU-level differences when predicting community responses to environmental disturbances such as coral bleaching. While generalist OTUs dominate natural communities, these do poorly in incubations, and we hypothesise that white light specialists may become key players during coral bleaching events. Our work is a substantial advance in our understanding of Ostreobium ecology and provides a framework for interpreting future environmental sequencing data, offering insights into the functional roles of the different OTUs.