Microbial Ecology

Myrmecocystus honeypot ants rely on specialized workers, repletes, to store dissolved carbohydrates in their crops long term. The repletes store this liquid, which does not spoil in their crops, for many months at a time. When resources are scarce, repletes redistribute the stored nutrients to their colony members via trophallaxis. While we suspect that the gut microbiome of honeypot ants may aid in spoilage prevention, before we can investigate this, we must first characterize it. Here, we used 16S rRNA gene sequencing to determine the microbial community composition across six Myrmecocystus honeypot ant species, sampling multiple colonies, castes, and organs. We found that microbiome community composition was strongly shaped by species, with variation between colonies in M. arenarius, M. depilis, and M. mexicanus. Organ level differences were observed in the crop and midgut in M. mexicanus. Caste differences were observed in M. flaviceps and M. mexicanus. Replete crops of M. mexicanus and M. depilis were enriched in Fructilactobacillus, other lactic acid bacteria, and acetic acid bacteria, whereas halophiles were more prominent in the gut of species such as M. flaviceps and M. wheeleri. In this study we demonstrate that Myrmecocystus ants host species-specific gut microbiomes and identify an association between lactic acid bacteria, acetic acid bacteria, and halophiles within replete crops. While much work remains in understanding the roles of the microbes in the symbiosis with their host ants, the dominance of these particular taxonomic groups suggests an association with a high sugar environment and a potential microbial role in preventing spoilage of the crop contents.

Characterizing the household bacterial microbiome allows for a stronger understanding of the various microbes that a person is exposed to everyday in their home. Exploring household microbiomes in different countries around the world increases - our understanding of the impact cultural differences might have on niche microbial communities in the house. The goal of this study was to use shotgun metagenomics to characterize the microbiome for ten locations around the home in ten different houses from three different countries (Mysuru, India; Dubai, United Arab Emirates (UAE); and Tucson, United States of America (USA)). There was a significant difference in alpha diversity between the three countries (ANOVA, p<0.05) with homes in Mysuru, India showing significantly higher bacterial diversity compared to Dubai, UAE and Tucson, AZ, USA. Beta diversity analysis of the homes found that bacterial communities significantly differed between cities (PERMANOVA, p<0.01) and within cities by household locations (PERMANOVA, p<0.001). Locations such as underneath the toilet rim, bathroom and kitchen sinks had the highest levels of bacterial diversity across the three cities compared to other sampling areas. A core microbiome of Actinomycetes and Gammaproteobacteria was found in all homes in all three cities. Within each city, a core microbiome was identified at the species level within specific household locations in each city. Over 90% of bacterial taxa found in the homes were a part of the human-associated phyla Actinomycetes (eg. genera Brevibacterium, Corynebacterium, and Microbacterium), Pseudomonadota (eg. genera Acinetobacter, Moraxella, Pantoea, Paracoccus, and Psuedomonas), and Bacillota (genus Streptococcus), which was comparable to previous studies. The household microbiome is variable in different locations in the house and on a global scale. Factors such as human activity, cultural practices, climate, and surface type and use may drive this diversity. Characterizing the household microbiome on a global scale allows for a better understanding of what drives microbial diversity, increasing our understanding of how microbial communities are shaped by the environment and how humans influence their dynamics, as well as any risks to human health that the built microbiome may potentially pose. Impact StatementThis research contributes to the understanding of the built microbiome, specifically how it varies within the house, within cities, and across the globe. This can aid in our understanding of microbial dynamics in environments with heavy human influence and help develop and improve hygiene habits and products which are mindful of the existing microbiome. Data SummaryDNA sequence data from this research is publicly available on the NCBIs Sequence Read Archive under BioProject PRJNA1416920. Data was analyzed using python and R code. Analysis protocols and information on software versions, packages, and more can be found within the text and in the following github repository: https://github.com/carolinescranton01/Global_Household_Microbiome. The authors confirm all supporting data, code and protocols have been provided within the article or through supplementary data files.

Despite their importance for individual fitness and population processes, the microbiota of many ecologically significant insects remains poorly explored. Even less is known about the interactions between microbial communities inhabiting insects and their surrounding environment. Ant infrabuccal pockets (IBPs), representing the interface between the digestive tract and the external environment, provide an opportunity to study these interactions. Here, we aimed to characterize ant-microbial interaction networks in the forest floor by profiling fungal and bacterial communities associated with the IBP of Formica polyctena ants, known as ecosystem engineers in temperate forests. We used direct microscopy, culturing, and sequencing amplicons of ITS1, ITS2, 18S rRNA marker regions to describe fungal communities, and 16S rRNA metabarcoding to characterize bacterial communities. Classical methods combined with a multi-marker amplicon sequencing allowed for a comprehensive description of the IBP microbiota. Fungal communities consistently contained representatives of 15 ecologically diverse genera, including insect-associated yeasts and primarily saprotrophic or endophytic fungi. Bacterial communities were dominated by genera previously reported from ant guts, mainly Bacilli and Alphaproteobacteria, and showed greater stability among ant colonies than fungal communities. Further studies on red wood ants IBP microbiota would enhance our understanding of their role in shaping ecological networks in forest ecosystems.

This study assessed the prevalence of antibiotic resistance genes (ARGs) and virulence factor genes (VFs), DNA viruses, and medically-relevant pathogens in three major cities around the globe - Mysuru (India), Dubai (United Arab Emirates), and Tucson (Arizona, United States of America). Ten households were sampled in each city, at ten sites in the bathroom, kitchen, and living spaces. The alpha diversity of ARGs significantly differed between household locations in each country (ANOVA, p<0.05) and beta diversity (dissimilarity) analysis showed a significant association between the ARGs and the geographic locations (PERMANOVA, p<0.01). A set of ARGs were found in every location across the dataset (the core ARG profile) included 25 different genes. The alpha diversity of virulence factors differed across the household locations within the three cities (ANOVA, p<0.01). The beta diversity of VFs was not well explained by geographic location or location within the household (PERMANOVA, p=0.129 (geographic), p=0.127 (household)). There were 341 unique VFs found in the study, but only 5 core VFs across the dataset. Bacterial pathogens detected across the household included Escherichia coli, Acinetobacter baumanii, Klebsiella pneumoniae, and more. The DNA (and bacteriophage) virome varied between countries and was more diverse in Tucson and Dubai (top viral families included Poxviridae and Orthoherpesviridae - two families which contain human pathogens - and Steitzviridae, a family of bacteriophages) compared to Mysuru, where nearly all viruses were a part of the Muvirus genus (a bacteriophage which contributes to horizontal gene transfer by phage transduction). ImportanceThe diversity of the built environment microbiome is not well characterized globally. Household occupants interact with the built microbiome on a daily basis, and the built microbiome may have the potential to influence human health. The presence of pathogens in the built environment and the key genes contributing to microorganism pathogenicity were investigated in this study, as information on this is lacking on an international scale. The diversity of ARG and VFs across the globe, as well as the presence of medically relevant pathogens within the house that were found in this study highlights the need to explore further the factors which influence the household microbiome, virome, and resistome. This may aid in identify how the build microbiome may be shaped by humans and influence human health. Impact StatementThis research contributes to the understanding of the built microbiome, specifically how it varies within the house, within cities, and across the globe. This can aid in our understanding of microbial dynamics in environments with heavy human influence and help develop and improve hygiene habits and products which are mindful of the existing microbiome.

The microbiota and microbiome associated with zooplankton remains rather understudied compared to other animal groups and/or taxa. The present study aimed at investigating the whole-body bacterial microbiota of Daphnia spp. in two contrasting Greek lakes, the shallow and hypertrophic Lake Koronia vs. the deep and mesotrophic Lake Vegoritida, including both egg-bearing and non-egg-bearing individuals. In both lakes, 2,060 bacterial operational taxonomic units (OTUs) were found, with 223 of them being conditionally rare (crOTUs) with low contribution even for the dominant phyla, with L. Vegoritida having more crOTUs than L. Koronia. The individuals microbiota had inconsiderable overlap with the surrounding water microbiota in both lakes. The two lakes showed significant differences in their Daphnia -associated microbiota. L. Koronia had richer OTUs and rather homogeneous bacterial communities, with higher occupancy. Overall, no significant differences in between the microbiota of egg-bearing and non-egg-bearing Daphnia individuals in both lakes. However, regarding the most important OTUs (miOTUs), the L. Koronia miOTUs were highly overlapped between the individuals with and without eggs, with only one missing from the individuals without eggs. In L. Vegoritida the individuals without eggs had only six miOTUs and while egg-bearing individuals had nine different ones; the two lakes had no shared miOTUs., considerable differences occurred.. A total of 27 miOTUs, was found and belonged to the Pseudomonadota, unclassified Bacteria, Cyanobacteria, Bacteroidota, Bacillota and Actinomycetota. Those miOTUs, where assignment to the genus level was possible, they were related to Cyanobium, Mucilaginibacter, Flavobacterium and Staphylococcus. This study showed that lake morphotype and ecological status can exert some impact on Daphnia-associated bacterial microbiota, with more pronounced effects on egg-bearing and non-egg-bearing individuals.

Bacterial interactions-whether positive or negative - are crucial for the functioning of microbial communities. Though bacterial interactions are mainly expected to be negative, the sign and strength of interactions are predicted to be context dependent, with interactions typically being more positive in more stressful and nutrient-poor conditions. However, systematic studies investigating how the environment affects interactions between multiple taxa are lacking. Here, we determine if interactions between a panel of natural soil isolates change in response to the environment in which they are grown, with two different artificial media used (one simple and one complex) and a more ecologically relevant soil wash. To maximise natural variation in interactions, we collected multiple isolates from multiple sites: co-occurring (sympatric) isolates were predicted to show more negative interactions than allopatric isolates because of greater overlap in resource use. Pairwise interactions were in general negative, but more negative when grown in a complex lab-derived medium (Tryptic Soy Broth). Mutually beneficial interactions were most common in a simple resource medium (M9 minimal media) and exploitative interactions were most frequent in a soil broth. These patterns were independent of whether species originated from the same or a different site. The study supports the prediction that nutrient rich environments promote more negative interactions, and that measuring interactions of soil isolates in standard lab media is likely to misrepresent interactions occurring in natural environments.

Ectomycorrhizal (EcM) fungi are well-recognized symbionts impacting tree health and ecosystem functioning globally, yet understanding of their timing of proliferation in soils across seasons and years remains limited. We analyzed monthly patterns of EcM fungal abundance and community structure over two years in five temperate monodominant forest plots via quantitative PCR and Illumina sequencing. We found that the phenological dynamics of EcM fungi differed significantly by host tree leaf habit, fungal exploration type, fungal genus, and soil moisture. Overall, total EcM fungal abundances based on qPCR consistently peaked in autumn, and were more dynamic in evergreen than deciduous plots, supporting ideas of surplus carbon and asymmetric above-belowground dynamics. Longer-distance exploration types peaked earlier and were more stable than shorter-distance types, suggesting an independent and supportive role in releasing spring nutrients. About half of 20 focal taxa consistently peaked in either autumn, summer, or spring, while others were either host- and/or year-dependent. Our findings highlight that phenology is a key EcM fungal trait best explained by both host and fungal contributions, and future studies across biomes should consider seasonal shifts and sampling to elucidate phenological traits. Summary- The timing of belowground production and seasonal community dynamics remain poorly understood for ectomycorrhizal (EcM) fungi. - We collected soils monthly for two years from five temperate monodominant forest plots. - Fungal production peaked in autumn, shorter-distance and evergreen-associated spanned wider ranges, and half of focal fungal genera showed seasonal preference, emphasizing autumn surplus carbon and spring nutrients from long-distance types. - Future studies should consider seasonal shifts when sampling EcM fungal communities, and forest carbon models should include asymmetric above-belowground phenology. Translated Summary (Spanish)- La fenologia de la produccion y composicion de comunidades de hongos ectomicorrizicos (EcM) es poco estudiada. - Recolectamos suelos mensualmente por dos anos de cinco parcelas mono-dominantes templados. - Produccion maxima de hongos ocurrio en otono, hongos asociados con arboles siempreverdes y de exploracion de corta-distancia observaron rangos mas amplios, y la mitad de generos de hongos focales observaron preferencia estacional, enfatizando extra carbono en otono y nutrientes en primavera de tipos larga-distancia. - Estudios deben considerar cambios estacionales para el muestreo de hongos EcM, y modelos de carbono deben incluir fenologia asimetrica entre hojas y hongos. Plain language summaryEctomycorrhizal fungi are critical for the global carbon cycle, but their seasonal and inter-annual growth patterns remain unclear. We sample soil DNA monthly over two years across five different monodominant temperate forest stands. We find an overall belowground peak in autumn, with significantly later growth under wetter conditions, more dynamism with evergreen trees, and distinct spring growth by longer-distance fungi.

Labyrinthulomycetes are a class of fungus-like heterotrophic protists from the Stramenopiles lineage, recognized for their ecological role as decomposers and contributors to nutrient cycling. They colonize various substrates, from seaweed to terrestrial environments, utilizing ectoplasmic networks for nutrient absorption. This study characterized a novel Labyrinthula strain associated with the marine diatom Biddulphia. Phylogenetic analysis of the full-length 18S rRNA gene positioned this strain as a new species, Labyrinthula merlionensis sp. nov. Scanning electron and light microscopy observations revealed bi-flagellated zoospores and spindle-shaped vegetative cells with ectoplasmic networks. Time-series observations of the interactions between L. merlionensis and Biddulphia were categorised into different phases: establishment, infection, and aggregation. Scanning electron and confocal microscopy observations during the infection phase established the use of ectoplasmic nets to target the marginal ridge regions between diatoms, and the detection of labyrinthulid cells within diatom frustules. These findings enhance the understanding of the diversity, morphology, and ecological roles of Labyrinthulomycetes, particularly their intra- and extra-cellular interactions with diatom hosts.

Most gut microbiome studies have focused on bacteria, leaving a knowledge gap regarding gut associated fungi. We assessed fungal diversity in the gastrointestinal tract of the reptilian order Testudines (turtles and tortoises) using samples from 6 families, 19 genera, and 27 species. A highly diverse community affiliated with 17 phyla and 157 orders was encountered, with four phyla (Neocallimastigomycota, Chytridiomycota, Ascomycota, and Basidiomycota) representing 89.13% of the community. Neocallimastigomycota was identified in host families Testudinidae (land tortoises), Chelidae, Chelydridae, Emydidae, Geoemydidae, and Kinosternidae, with higher relative abundances in Testudinidae (40.18{+/-}37.97%) compared to all other families combined (2.71{+/-}4.04%). Neocallimastigomycota sequences were mostly affiliated with orders Testudinimycetales in the host family Testudinidae and Neocallimastigales in other host families. Chytridiomycota was identified in all host families, but was more ubiquitous and abundant in Kinosternidiae (45.17{+/-}34.12%), and exhibited a high level of variability across samples. Dikarya communities were highly diverse, with 108 orders identified, and, similar to Chytridiomoycota, exhibited a highly stochastic distribution pattern. Representatives of multiple yet-uncultured phyla (Candidatus "Algovoracomycota", "Sedimentomastigomycota", "Tartumycota" and "Cantoromastigomycota") were identified, as well as eight novel orders in Chytridiomycota and Rozellomycota. Deterministic selection shaped community assembly in the host family Testudinidae, while the process was more stochastic in other host families. Distinct community structure was driven by differences in abundance and identity of the Neocallimastigomycota when comparing Testudinidae to. Our results describe a diverse and dynamic fungal community, shaped by the co-occurrence of autochthonous (resident) and transient (allochthonous) members of the gut microbiome. ImportanceFungi are known to inhabit the gastrointestinal tract (GIT) of humans and mammals. However, information on the fungal community in the GIT of reptiles is relatively sparse. We investigated the diversity and community structure of fungi in the reptilian order Testudines. We conducted a culture-independent diversity survey on fecal samples obtained from 27 different host species. We identify representatives of 17 fungal phyla. As well, we demonstrate that the anaerobic gut fungi (phylum Neocallimastigomycota) are not restricted to the family Testudinidae (land tortoises) as previously suggested, but could successfully colonize and inhabit all other testudines families, including those exhibiting a predominantly omnivorous or carnivorous lifestyles. In addition, we expand on the known fungal diversity by identifying additional representatives of multiple recently described yet-uncultured phyla, and describe multiple novel orders and classes within existing phyla. Collectively, this effort adds to the growing body of knowledge of mycobiomes in underexplored animal hosts.

Plants recruit microorganisms to form mutually beneficial associations that enhance their health, productivity, and resilience. The composition of the plant microbiome is shaped by factors such as host species, developmental stage, genotype, and tissue type, with microbial recruitment mediated by plant exudates and secondary metabolites. Eugenia uniflora, a Myrtaceae species native to Brazils Atlantic Forest, produces pharmacologically relevant secondary metabolites and holds ecological and economic value. However, little is known about its associated microbiome, particularly from a metagenomic perspective. In this study, we investigated the phyllosphere bacterial communities, both epiphytic and endophytic, of E. uniflora across two developmental stages (young and mature trees). We also examined the core microbiome shared between E. uniflora and other Myrtaceae genera to better understand microbial diversity and structure within this family. Amplicon sequencing of the V3-V4 region of the 16S rRNA gene was conducted on 19 E. uniflora samples and 13 additional samples from three other Myrtaceae genera. In E. uniflora, we identified 1,456 bacterial ASVs representing 17 phyla, 115 families, and 171 genera. Alpha and beta diversity analyses revealed significant differences in bacterial community composition between developmental stages. Genera such as Massilia and Hymenobacter were more abundant in mature trees, while Aureimonas and Terriglobus were more common in young plants. Leaf microbiome functional potential shifted with plant age, with older leaves favoring secondary metabolite production and younger leaves emphasizing microbial interactions and defense. A total of 16 genera formed the Myrtaceae core microbiome, with five, Methylobacterium-Methylorubrum, Hymenobacter, Sphingomonas, Bdellovibrio, and Terriglobus, present in 100% of samples. Notably, [~]0.7% of the bacterial diversity remained poorly classified, highlighting the underexplored nature of Myrtaceae-associated microbiomes and their potential for bioprospecting.

Microbial communities are central to the biogeochemical cycling of nutrients, critically shaping ecosystem functioning and influencing climate change mitigation. Mangrove ecosystems are among the most important global carbon sinks that enable large amounts of carbon to be sequestered and stored. However, gaps persist in understanding the fundamental aspects of microbial-driven carbon cycling in these environments. This research explores the microbial taxonomic and functional diversity related to carbon cycling in selected tropical mangrove sediments across various locations and depths. Sequencing data analyses based on the 16S rRNA gene revealed distinct microbial community composition but conserved predicted functions across the different mangrove locations. Depth was a strong influence on the functional composition, with carbon-related pathways and metabolic strategies differing between top and bottom sediments. Putative functional gene abundance analyses revealed that carbon fixation processes were among the top carbon-related pathways, suggesting the key role of mangrove microbial communities in sustaining long-term carbon storage. Within these communities, Desulfobacterota appeared as a primary contributor to carbon fixation, while Chloroflexota played a significant role in carbon metabolism and methane cycling. Co-occurrence network analyses also revealed that these microbial groups were among the keystone taxa in mangrove sediments. Our study adds on to the body of knowledge on the mangrove microbiome and their carbon metabolic processes, which helps to improve strategies for managing and leveraging these vital carbon sinks.

Holometabolous polyphagous insects undergo complete metamorphosis and exploit multiple host plants, exposing them to highly variable ecological conditions across both life stages and host plants. Whether these species, like specialist ones, harbour a stable core microbiota, or whether life stages or host plants act as the primary drivers of microbiota assembly remain open questions. Here, we characterized the fungal and bacterial communities associated with Drosophila suzukii across life stages and host fruits using 16S and ITS metabarcoding. We tested the relative influence of life stage and host fruit on microbiota composition, using community and network-based analyses. We first identified that host fruit significantly structured fungal communities, but not bacterial ones. Yeast communities were rather fruit-specific: Hanseniaspora and Pichia mostly associated with cherries and strawberries, contrary to Metschnikowia with blackberries. In contrast, bacteria and filamentous fungi were shared across fruits, constituting for fruits a core microbiota dominated by Gluconobacter cerinus, Tatumella and Cladosporium. Second, we found that both bacterial and fungal D. suzukii communities were structured by life stage, and that fungal, but not bacterial communities, were also structured by host fruits. D. suzukii individuals harboured a core bacteria composed of G. cerinus and a niche-specific microbiota composed of yeasts: Hanseniaspora typical in individuals related to cherry and strawberry, and Metschnikowia to blackberry. Components of both core and niche-specific microbiota were most likely horizontally acquired by D. suzukii from host fruits. Taken together our results underline the importance of meta-community approaches to investigate tripartite interactions among insects, host plants and microbiota. IMPORTANCEThe role of gut microbiota in mediating interactions between phytophagous insects and their host plants has been well illustrated in specialist species. However, it has been less comprehensively studied in polyphagous species, which infest multiple host plants, and across life stages for holometabolous species experiencing separate ecological niches through development. We tested the existence of a core, a niche-specific and a stage-specific microbiota in a polyphagous holometabolous species, D. suzukii. We examined both fungal and bacterial communities in larvae, pupae and emerging flies infesting three host fruits. Our results showed first that the assembly of bacteria, filamentous fungi and yeasts on fruits is driven by different ecological processes. Second, that D. suzukii harbours a core bacterial microbiota, a niche-specific microbiota constituted by yeasts and no stage-specific microbiota. Our study emphasizes the importance of considering jointly the assembly of host plant and polyphagous insect microbial communities to better understand the ecology and evolution of insect-microbe interactions.

The fish skin microbiome serves as a protective barrier, influencing host health and facilitating interactions between the host and its environment. While several studies have characterised the composition and roles of the fish skin microbiome, there remains a paucity of data on how environmental variation influences these microbes in natural populations. Here, we used 16S rRNA gene sequencing to characterise the skin microbiome of wild three-spined stickleback populations and examine how environmental factors influence microbial diversity and community composition across 17 freshwater lochs on the island of North Uist, Scotland. Analysis of 239 samples revealed a set of dominant bacterial genera commonly associated with other fish species, including Janthinobacterium, Pseudomonas, Acinetobacter, and Psychrobacter, that constituted a core skin microbiota across lochs. Microbiome composition was primarily shaped by environmental variables, particularly habitat, water pH, conductivity, and metal concentrations, with pH emerging as a key driver of community structure. Host sex also influenced microbiome variation, with several taxa differing in relative abundance between males and females. Alpha-diversity was higher among stickleback fish from lochs with a neutral pH compared with those from alkaline and acidic environments. Differential abundance analyses identified 27 and 24 amplicon sequence variants (ASVs), respectfully, associated with variations in pH and host sex, including members of Psychrobacter, Sphingobacterium, Carnobacterium, Chryseobacterium, and Arthrobacter, highlighting the combined influence of environmental and host factors on microbiome composition in wild fish populations in freshwater environments.

O_LIPhyllosphere microbiomes are subject to microbial import from various sources and undergo substantial changes during phenological changes of plants. However, these processes are still poorly understood for forest canopies. We propose that phenology-driven changes in host properties, and rainwater-mediated, within-canopy transport shape the phyllosphere microbiome in temperate forests. Leaves and throughfall samples were collected from oak, ash and linden trees at top, mid, and bottom canopy positions at the Leipzig canopy crane facility (Germany) at time points representing early, mid and late phenological stages. Bacterial community composition was assessed by 16S rRNA gene amplicon sequencing. C_LIO_LIPhenological stages explained 19% of phyllosphere bacterial community variation, followed by tree species identity (12%) and canopy position (2%). Later phenological stages exhibited more homogeneous and functionally redundant phyllosphere communities along with a strong decline of plant pathogens and increasing potential for microbially mediated biocontrol mechanisms. Throughfall transported up to 1011 bacterial cells per litre with maximum bacterial fluxes at the canopy top. C_LIO_LIOur findings demonstrate that in temperate forests, phenology-driven effects on the phyllosphere microbiome are far more important than tree species specific effects. Extent and selectivity of throughfall-mediated mobilization may play a crucial role for the spatial heterogeneity of microbial communities in tree crowns. C_LI

BackgroundBuilt environment microbiome studies have identified numerous factors that shape indoor microbiomes, yet the reproducibility of these findings across buildings, timepoints, and research groups remains unclear. Differences in sequencing protocols, sampling design, and environments pose major challenges for cross-study comparisons, particularly in low-biomass environments where technical variation can obscure biological signal. To address this gap, we constructed a simple ontology which groups samples into one of three categories: hand, hand-associated surfaces, and floor then applied it to four publicly available 16S rRNA gene datasets: a hospital, university dormitory, Air Force dormitory, and private residential houses. ResultsWe identified strong and reproducible separation between floors and surfaces with frequent human contact. We found that floors consistently harbored soil-associated taxa, including KD4-96, 67-14, Skermanella, and Sphingobacterium, whereas hands and hand-associated surfaces were enriched with skin-associated genera, such as Lawsonella and Cutibacterium. Within studies, these results were generally consistent across timepoints. Across studies, mixed-model PERMANOVA analysis revealed significant clustering by sample type, with modest effects of study, suggesting that biological signal outweighed differences in laboratory or sequencing methods. Leave-one-study-out random forest models achieved high AUCs for hand vs. floor comparisons (0.865 to 0.921), moderate AUCs for hand-associated vs. floor comparisons, and weaker performance for hand vs. hand-associated comparisons. Application of the batch-correction method DEBIAS-M did not improve effect sizes or classification performance, indicating that reproducible structure was already discernible without batch adjustment. ConclusionsDespite substantial temporal and environmental heterogeneity among studies, we found that the built environment microbiome has a reproducible bacterial signal. There was consistent enrichment of soil-derived taxa on floors and human-associated taxa on hands and hand-associated surfaces suggesting a stable microbiome despite differences in building type, occupancy, and methodology. These findings establish an important foundation for future studies, suggesting cross-study comparability, the accuracy of ecological inference, and the ability to support the development of predictive applications in indoor microbiome research.

Spartina alterniflora, the dominant plant species in salt marshes along the Atlantic and Gulf of Mexico coastlines of the Americas, is affected by disease and sudden vegetation dieback. Despite the foundational role of S. alterniflora in low-elevation salt marshes, the response of the native leaf-associated microbiome (i.e., phyllosphere microbiome) to leaf damage resulting from disease and environmental stress has not been explored. We hypothesized that healthy and damaged plants would show differentiation in their phyllosphere microbiomes following primary infection or exposure to environmental stressors. Here, we analyzed changes in prokaryotic and fungal relative abundance, diversity, and community composition in the S. alterniflora phyllosphere microbiome. We compared natural marsh and greenhouse plants in Georgia and South Carolina, USA, and collected leaves from healthy and damaged natural plants across two contrasting Spartina phenotypes that differ in their exposure to environmental stress. Our results show that plant origin (i.e., greenhouse vs. natural marsh), plant health status (i.e., healthy vs. damaged), and plant phenotype (i.e., short vs. tall Spartina) affect microbial relative abundance, alpha diversity, and community composition in the S. alterniflora phyllosphere. Damaged leaves presented higher microbial abundance and alpha diversity than healthy leaves, suggesting microbial proliferation following leaf damage. Plants raised from seeds in the greenhouse presented the lowest microbial abundance and Shannon diversity for both prokaryotic and fungal communities, indicating that in natural ecosystems the phyllosphere microbiota is acquired predominantly through horizontal transmission from the environment. Overall, this study provides novel insights into the assembly of the S. alterniflora phyllosphere microbiome. ImportanceSalt marshes are tidally influenced coastal wetlands that provide a range of ecosystem services to global and local communities, including protection from storm surge, water purification, and carbon sequestration. Spartina alterniflora is the dominant plant species in Atlantic and Gulf of Mexico marshes within the Americas. Fungal disease and exposure to environmental stressors have previously been described in marsh ecosystems and linked to extensive and sudden vegetation dieback in the southeastern U.S. In this study, we show that microbial proliferation follows plant damage caused by either fungal disease or environmental stress, leading to a profound change in native leaf-associated microbiota abundance, diversity, and composition (i.e., leaf microbiome dysbiosis). Using greenhouse plants as a control, we also demonstrate that microbes colonizing marsh leaves are acquired predominantly from the environment. Overall, this study advances our understanding of the leaf-associated microbiome of S. alterniflora, with implications for ecosystem management and restoration practices.

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.

The evolutionary divergence between Henophidia (non-venomous) and Caenophidia (venomous) snakes has produced distinct cranial morphologies, digestive strategies, and presence of specialised venom systems in Caenophidia, yet the extent to which these long-standing diverging trajectories have shaped cloacal microbiota assembly remains poorly understood. We characterised cloacal microbiota in 70 captive snakes (52 Caenophidia, 18 Henophidia) by 16S rRNA amplicon sequencing. Beta diversity was tested by PERMANOVA, differential abundance by ANCOM-BC2, community types by Dirichlet Multinomial Mixture modelling (DMM), and microbial interactions by SparCC co-occurrence networks. Predicted functional potential (PICRUSt2) was analysed by ALDEx2 differential abundance testing and elastic net feature selection. Henophidia exhibited significantly higher bacterial richness and greater compositional variability than Caenophidia. Community composition showed clade-associated differences (PERMANOVA) and partitioned into two distinct DMM community types. The Henophidia network was 11.9-fold denser and more modular, with Burkholderiaceae as a keystone hub, whereas the Caenophidia network was sparse. Henophidia showed predicted enrichment in C1 metabolic pathways (ethylmalonyl-CoA, formaldehyde assimilation I, glycine betaine degradation I, methylaspartate cycle), aromatic compound catabolism, and nitrogen recycling, whilst Caenophidia showed enrichment in allantoin and glucuronate degradation. This multi-method analysis suggests Burkholderiaceae as a candidate keystone taxon in Henophidia and indicates that phylogenetic clade is a major contributor to cloacal microbiota structure. The lower richness in Caenophidia raises a testable hypothesis that broad-spectrum antimicrobial activity of their venom components may selectively filter susceptible microbial lineages, motivating future shotgun metagenomic studies in wild populations of snakes.

Ectomycorrhizal (ECM) fungi are well-known for their crucial roles in forest health and productivity, yet their responses to various forest management practices are understudied, particularly in oak-dominated forests. The purpose of this study was to better understand the effects of silvicultural treatments on the diversity and community composition of ECM fungi in an oak-hornbeam forest in northern Hungary. We analyzed ITS2 rDNA metabarcoding data of soil-borne fungi to compare richness and community composition of ECM fungi among forest treatment types (clear-cutting, gap-cutting, preparation-cutting, tree retention in clear-cut areas, and control) and between sampling years (2020 and 2021). We found 268 ECM fungal genotypes, with the most diverse phylogenetic clades being /russula-lactarius (52), /tomentella-thelephora (47), /inocybe (40), /sebacina (27), and /cortinarius (20). We found significant compositional difference of ECM fungi among silvicultural treatments in both years, with some variations in richness. There were also small, but still significant compositional differences between the two years. Treatment effect was partly explained by altered environmental variables, such as relative humidity and soil temperature. These results highlight the importance of forest structure and the abiotic environment in driving community dynamics of plant-symbiotic fungi, with potential implications for forest health and productivity.

This study investigates the seasonal dynamics of the microbiome within the marine sponge Halichondria panicea from Baltic coastal waters, focusing on its symbiotic relationship with Candidatus Halichondribacter symbioticus. Over 16 months, we observed distinct summer and winter microbial communities, transitioning rapidly between these states during spring and fall. Marine sponges host complex microbiomes composed of diverse microbial taxa that play critical roles in host metabolism and nutrient cycling within marine ecosystems. While our understanding of sponge microbiomes has traditionally been based on static characterizations, the temporal dynamics of these associations across seasonal cycles remain poorly understood. In this study, we investigated temporal variation in bacterial symbionts of Halichondria panicea over 16 months in Baltic coastal waters using high-throughput amplicon sequencing of bacterial 16S rRNA gene sequences. The microbiota of H. panicea exhibited host-specific structure and a high degree of stability across seasons, despite fluctuations in environmental factors such as temperature, salinity, photoperiod intensity, and inorganic nutrient availability. In contrast, bacterial communities in surrounding seawater displayed large seasonal shifts which potentially mix with the sponge bacterial community, suggesting that different degrees of ecological pressures act on free-living and symbiotic marine bacteria. These findings establish an empirical baseline for identifying abnormal shifts in symbiont communities, which could be indicative of environmental stress or biological disturbance events.