Animal Microbiome

The teleost gastrointestinal tract hosts metabolically active microbial assemblages that play critical roles in nutrient use, immune regulation, physiology, behavior, growth, reproduction, and health status. However, the extent to which aquaculture production systems restructure these communities remains poorly defined. In this study, metagenomic sequencing was applied to characterize gut microbiota in largemouth bass (Micropterus salmoides) reared under four representative farming conditions: industrial aquaculture (IA), captive pond-based farming (CA), ordinary pond aquaculture with healthy fish (PH), and ordinary pond aquaculture with diseased fish (PD). Results revealed that the four groups exhibited pronounced differences in microbial diversity, taxonomic composition, functional capacity, carbohydrate-active enzyme profiles, and antibiotic resistance genes (ARGs). Notably, bacterial taxa accounted for more than 95% of the gut microbial communities across all groups. IA fish showed greater microbial richness and diversity than CA and PH fish, consistent with a more complex and potentially more stable intestinal ecosystem under industrialized rearing conditions. Taxonomic profiling at both the phylum and genus levels identified production system-specific microbial signatures. In particular, opportunistic pathogens, including Aeromonas veronii and Aeromonas hydrophila, were disproportionately enriched in CA fish, suggesting a microbiota profile associated with increased disease vulnerability. Functional prediction analysis further indicated distinct metabolic states among groups, with purine and fatty acid metabolism enriched in PD fish, potentially reflecting disease-associated disruption of host-microbiota interactions. Carbohydrate-active enzyme analysis revealed substantial variation in microbial carbohydrate-processing potential, whereas ARG profiling indicated that farming conditions were linked to distinct gut resistome patterns. Collectively, these findings identify aquaculture regimes as a key ecological driver of intestinal microbiome structure in largemouth bass and provide a mechanistic basis for optimizing microbiome-informed health management in aquaculture systems. Keywords: Largemouth bass (Micropterus salmoides); gut microbiota; aquaculture modes

Domestic dogs may influence human microbiota and act as reservoirs or sentinels for zoonotic bacteria, yet little is known about these dynamics in Southeast Asia. Using full-length 16S rRNA gene sequencing from rectal swabs, we analyzed microbiota from dog-owner pairs from four villages of rural Thailand (Nan province) to assess microbial diversity, shared taxa, and zoonotic potential. Dogs exhibited higher microbial and putative pathogen diversity than humans, particularly during the rainy season. While dog-owner pairs shared more bacteria than unrelated individuals, overall overlap remained limited - likely driven by their dogs free-roaming lifestyles and broader environmental exposures. However, co-occurrence of putative zoonotic pathogens such as Shigella spp. and Klebsiella pneumoniae suggests dogs could serve as useful indicators for specific public health risks, with further targeted pathogen screening needed to validate these results. Our findings highlight the value of integrating dogs into zoonotic surveillance programs, particularly in regions with free-ranging dog populations and close human-animal contact.

Cattle harbor diverse bacterial communities across the gastrointestinal, respiratory, reproductive, mammary and other anatomical systems, but body-wide microbial biogeography remains poorly defined. To address this knowledge gap, we conducted a meta-analysis of publicly available bovine 16S rRNA gene amplicon sequencing data from 5,637 samples from 47 studies across six geographic regions and 27 anatomical sample types. Bacterial community structure differed significantly among sample types (PERMANOVA, R2 = 0.245, P = 0.0001), indicating spatial organization of bacterial communities across the bovine body, although study-level effects also contributed substantially to community variation. Communities were generally more similar within than between anatomical systems. Bacterial richness, diversity, taxonomic composition, and indicator taxa varied among sample types, with gastrointestinal, mammary-associated, ocular, and hoof microbiota exhibiting greater diversity than microbiota from liver, joint, and several reproductive samples. Distinct bacterial communities characterized the gastrointestinal, respiratory, reproductive, and mammary systems, as well as other anatomical sites. Despite these differences, several bacterial taxa were shared across multiple anatomical niches, particularly among male and female reproductive sites and among mammary-associated niches. This study provides a comprehensive body-wide characterization of bacterial biogeography in cattle and establishes a baseline for future studies of bovine microbial ecology and host-microbiome interactions.

Gut microbiome responses to anthropogenic disturbance vary across wildlife species, even within similarly disturbed landscapes. What drives this variation is unclear: whether it reflects anthropogenic exposure itself or broader ecological differences among hosts. We tested this using three macaque species with contrasting ecology, Bonnet, Rhesus, and Lion-tailed macaques, sampled across 12 sites in southern India spanning contrasting anthropogenic exposure, using 16S rRNA gene sequencing (n = 127) and shotgun metagenomics on a subset of samples. The two synurbanized species exhibited a similar magnitude of microbiome restructuring but differed in the taxa underlying these changes; no differentially abundant amplicon sequence variants were shared across all three species, indicating that shared anthropogenic exposure did not produce uniform microbial responses across hosts. The specialist Lion-tailed macaque showed a more extensive response, characterized by reduced diversity and phylogenetically structured compositional change. The Bonnet macaque showed greater microbial similarity with the Rhesus macaque than with the Lion-tailed macaque during sympatric co-occurrence. Despite taxonomic divergence, functional pathway architecture was broadly conserved across species and habitats, with selective shifts in pathways including vitamin B6 biosynthesis and fermentation. Together, these findings show that microbiome responses to anthropogenic environments are jointly shaped by ecological context and host ecology, with host differences in diet, habitat use, and ecological history influencing the magnitude and nature of microbial restructuring. These findings show that taxonomic diversity and functional potential respond as partially decoupled axes under anthropogenic pressure, with implications for assessing microbiome resilience across ecologically heterogeneous wildlife.

This study was carried out on two rabbit lines, each selected for a feed efficiency criterion (growth on restricted feed for the AlimR line or residual consumption on an adlibitum diet for the ConsR line), introduced into a facility by adoption of newborn kits by SPF (specific pathogen free) females. Following this break in symbiotic transmission, fecal samples from females were used to study the microbiota composition of the two lines over seven generations (17 females per line and per generation). The proportion of Firmicutes was significantly higher in the AlimR line than in the ConsR line (82.22% vs 78.21%, p<0.0001). Conversely, the proportion of Bacteroidota was significantly higher in the ConsR line than in the AlimR line (20.38% vs 16.50%, p<0.0001). The effect of generation was also significant for the abundance of Firmicutes and Bacteroidota (p=0.0009 and p<0.0001, respectively). The Shannon diversity index was significantly higher in the AlimR line than in the ConsR line (5.23 vs 5.14, p=0.02). The effect of generation was also significant for the Shannon index (p=0.003). Finally, a discriminant principal component analysis of the microbiota composition allowed us to distinguish both the two lines and the generations, in particular the 11th selected generation (animals adopted by SPF females) and the 18th generation.

Wildlife habituation across the globe poses threats to human and animal safety and alters regular ecological function, yet the wildlife, environmental, and public health ramifications are poorly characterized. The gut microbiome is a valuable proxy for these factors as it influences host health, is affected by anthropogenic exposure, and may contribute to the environmental spread of virulence factors. Despite their importance, wildlife gut microbiomes are largely uncharacterized, both in composition and function. We characterized American black bear (Ursus americanus) gut microbiomes across a spectrum of lifestyles including wild (conflict-free), habituated (conflict-prone), and captive individuals. 16S rRNA gene sequencing was used for compositional analyses and whole genome short-read sequencing (WGS) was performed on a subset of bears to investigate microbiome function and virulence factors. We found that habituated, wild, and captive bears have unique gut microbiome compositions that are indicative of their respective lifestyles. In particular, captive bears are enriched in carnivory-associated taxa and habituated bears in inflammatory taxa. WGS reveals wild bears have increased fungi and soil-associated bacteria, as well as higher proportions of unclassified reads. Habituated bears have significantly increased levels of antimicrobial resistance genes, and captive bears are enriched in enterotoxins. Our results demonstrate previously undescribed wildlife physiological and One Health consequences of habituation.

Traditional veterinary pathology investigations on wild animals generally conclude a primary cause of death based on a combination of observed clinical signs, gross and microscopic lesions, microbial culture and PCR. However, these methods may fail to capture the broader microbial community of the host. We employed metatranscriptomic sequencing to characterise the potential pathobiome of necropsied marine turtles, comparing these findings with corresponding veterinary pathology reports. Tissue samples were collected from 76 individuals spanning three species of marine turtles -- the green turtle (Chelonia mydas), the hawksbill turtle (Eretmochelys imbricata), and the loggerhead turtle (Caretta caretta) -- across the coastline of New South Wales, Australia. Reported causes of mortality ranged from infectious disease to trauma. We identified a diverse range of parasites within the genera Neospirochis, Hapalotrema, Paraorgymatobothrium, Eimeria and Callitetrarhynchus, and bacteria including the genera Aeromonas, Clostridium, and Vibrio. Novel viruses of the families Astroviridae, Caliciviridae, Hepaciviridae, Hepeviridae, Nyamivirdae, Orthomyxoviridae, Picobirnaviridae and Rhabdoviridae were also identified. In cases where infectious disease was noted in the pathology report, we frequently observed high pathogen read counts consistent with the listed primary diagnoses. However, we also detected a range of incidental infections involving eukaryotic parasites, bacteria, and viruses. The diversity of infectious agents detected in marine turtles suggests that the observed disease outcomes are unlikely to be driven by a single causative agent, but rather by complex interactions between host health, the pathobiome, and environmental stressors. We discuss the potential implications for turtle health and conservation.

Host genetic markers often fail to resolve regional origins in highly connected or panmictic marine species. The Japanese jack mackerel, Trachurus japonicus, is a commercially important fishery species around Japan that shows little or no detectable population structure. Here, we used nationwide multi-omics profiling to compare host genomic variation and gut microbiome composition in wild T. japonicus collected from coastal regions across Japan. We generated MIG-seq data for 43 individuals and 16S rRNA gene profiles for 24 individuals; after quality filtering, 19 individuals remained for matched host-microbiome comparison. Genome-wide host SNP analyses showed weak or absent geographic population structure, consistent with previous evidence of panmixia in Japanese waters. In contrast, gut microbiome composition showed geographic structuring based on Bray-Curtis dissimilarity and PERMANOVA, and this pattern was not explained by proximity to river mouths or host-related variables. Locality- or individual-associated bacterial lineages contributed to the observed differences in the microbiome, while chloroplast-associated and Cyanobacteria-assigned ASVs suggested recent dietary or environmental input. These results indicate that gut microbiome can show regional biological variation not apparent from host genetic markers alone. Our study provides a proof-of-concept example of integrating host genomics and gut microbiome profiling to evaluate regional characteristics and origins in highly connected marine animals.

Background: Microbiome research has linked microbial community structure to host phenotype, but translating microbiome-based discovery into defined interventions with reproducible long-term host effects remains challenging. We evaluated whether a defined four-member consortium of rumen native microorganisms identified through associations with host phenotype could influence herd retention. Cull outcomes (sold or died vs. retained in herd) were analyzed across 20 controlled commercial dairy trials encompassing more than 40,000 cattle housed across 110 experimental pens, with each trial conducted for at least 5 months. Treatment pens received a defined daily dose of Galaxis Frontier (GF, Native Microbials, San Diego, CA), containing Pichia kudriavzevii, Clostridium beijerinckii, Ruminococcus bovis, and Butyrivibrio fibrisolvens, incorporated into the ration; control and treatment groups were otherwise managed equivalently. Cow-level time to culling was modeled within each trial, accounting for pen-level clustering, and trial estimates were combined using random-effects meta-analysis. Results: Daily treatment with rumen native microbes was associated with a 9% lower hazard of culling across 20 trials (hazard ratio = 0.91, 95% confidence interval: 0.86-0.96; P < 0.001). Between trial heterogeneity was negligible, and treatment effects were not significantly moderated by breed or geographic region. Detailed health records from one trial were examined to further characterize health outcomes associated with GF; treatment was associated with fewer transition-related disorders, particularly ketosis and metritis, whereas mastitis and lameness were largely unaffected. These findings suggest that the health effects associated with GF may be more pronounced for disorders linked to physiological and metabolic adaptation during the transition period than for health events more broadly. Conclusion: Across diverse commercial dairy environments, treatment with a defined consortium of rumen native microbes was associated with improved dairy cow retention. The consistency of this association across independent trials provides large scale evidence that targeted application of native microorganisms can translate into reproducible, long-term outcomes for the host. Complementary health event data suggest that improved transition period health and metabolic stability may contribute to this effect. These findings support the potential for microbiome targeted interventions to influence productive longevity under commercial conditions.

The gut microbiota is a key player in energy balance, impacting both digestion efficiency and the production of metabolites involved in metabolism. Its composition is highly adaptable, especially in response to diet. Changes in human diet and lifestyle over time - from active, fibre-rich diets to sedentary habits with calorie-dense foods - have likely contributed to the rise in metabolic diseases. Rodent models are widely used to study the links between diet, microbiota and metabolism. However, they have important limitations (e.g. artificial environments, uniform diets and biological differences from humans) which can affect the translation of findings to humans. While mice and humans differ in their microbiota species, they do share some functional similarities. The grey mouse lemur (Microcebus murinus) has been proposed as a promising alternative model. This small primate experiences strong seasonal changes in food availability, leading to distinct physiological states (energy-saving in winter vs active in summer), even in captivity. It is increasingly recognized as a valuable model for biomedical research, supported by recent genomic and molecular advances. However, its gut microbiota has not yet been the subject of study. Consequently, the present study focuses on investigating the gut microbiota of the grey mouse lemur, with a particular emphasis on how these microbiota vary under different dietary regimens. The microbiota of animals fed the standard colony diet was dominated by Prevotella, Bifidobacterium, Megamonas, Streptococcus, Megasphaera and Lactococcus, showing an Prevotella driven enterosignature. We showed that switch from a classical control diet to 3 different diets resulted in change on microbiota composition that is associated with functional redundancy. The present work underline the interest of Microcebus murinus as model for diet and lifestyle studies in relationship with metabolic diseases.

Histomonosis, caused by Histomonas meleagridis, is a severe disease in poultry with main pathological lesions in ceca and liver, leading to significant animal suffering and economic losses, with no licensed treatments available. While cecal pathology and microbiota disruption have been documented, the metabolic consequences of infection and vaccination remain poorly understood. Here, we used untargeted LC-MS metabolomics to profile cecal tissue and luminal digesta in a controlled vaccination-challenge chicken trial (n=100, 25 birds/group), including uninfected and unvaccinated but challenged controls, together with birds vaccinated either cloacally, or orally via an edible gel following challenge, across five post-challenge timepoints. Results showed that H. meleagridis infection caused extensive, time-dependent disruption of the cecal metabolome, especially at 7-14 days post-infection (DPI). At DPI 14, 325 tissue and 359 digesta metabolites were significantly altered in unvaccinated challenged birds. Infection was associated with depletion of microbiota-related metabolites, including dicarboxylic acids, tryptophan indoles, phytochemical derivatives, bilirubin catabolites, phenolics, vitamins, and secondary bile acids, alongside accumulation of microbial substrates and fermentation products. Changes in the host-associated metabolites indicated mucosal inflammation, epithelial injury, mitochondrial and amino acid metabolic disturbance, oxidative stress, and membrane lipid remodeling. Cloacal vaccination provided greater metabolic protection than oral vaccination, with 78.3% of disrupted metabolites preserved at DPI 14, compared with 60.7% vaccination by oral gel delivery, pointing towards the importance of vaccine take. By DPI 21, cloacally vaccinated birds showed nearly complete metabolic recovery across most pathway groups, whereas gel-vaccinated birds retained broader metabolic disturbance. These findings reveal histomonosis as a disruption of the cecal host-microbiota metabolic interface and provide a metabolite-level framework for future studies of pathogenesis, vaccine development, and prevention strategies.

While the influence of microorganisms in the thermal physiology of animals is increasingly recognised, the role of the gut microbiome in enhancing phenotypic plasticity remains underexplored, particularly in ectotherms. Here, we tested the effect of gut microbiomes in modulating the physiological response of lizards to sudden temperature fluctuations. In captivity, we conducted an experiment with wild-caught Podarcis lizards: Mediterranean Podarcis liolepis (n=9) adapted to warm and dry environments, and Atlantic P. muralis (n=18) adapted to cooler and wetter conditions. Animals were assigned to three groups (intervention, control-cold and control-warm) and subjected to microbiota manipulations using antibiotics and faecal microbiota transplants (FMT). In the intervention group, cold-adapted P. muralis received faecal suspension from warm-adapted P. liolepis, while control-cold and control-warm groups received intraspecific transplants. Microbiome variation was analysed through genome-resolved metagenomics of 162 faecal samples, yielding 313 MAGs across 15 phyla, containing almost 1M annotated genes. One week after transplantation, microbiomes of control animals differed from their original state, whereas those of the intervention group (P. muralis), transitioned to a state resembling P. liolepis. After two weeks, both control groups showed trends of reversing their microbiomes to their original states. However, the intervention group displayed a distinct trajectory, resembling the microbial donors'. Although potentially relevant trends were observed, these microbiome alterations did not translate into detectable significant differences in their thermal physiology. Nevertheless, our results provide proof of concept for microbial transplants in lizards, and opportunities to further explore the link between gut microbes and host physiological responses in ectotherms.

Background: Bacterial blotch is a major disease of cultivated white button mushroom (Agaricus bisporus) traditionally attributed to individual Pseudomonas pathogens. However, the recurrent detection of diverse bacterial taxa in blotch-affected mushrooms suggests that disease may involve broader changes in microbial community organization. We characterized bacterial communities associated with symptomatic and asymptomatic mushrooms and examined bacterial succession in the casing layer across early, pinning, and harvest stages at two commercial mushroom farms in the United States using complementary 16S rRNA gene amplicon sequencing and shotgun metagenomics. Results: Mushroom-associated communities were dominated by Pseudomonas regardless of disease status, indicating that bacterial blotch was not simply associated with increased abundance of the dominant genus. Instead, symptomatic mushrooms exhibited significant community restructuring, enrichment of specific taxa, and markedly reduced microbial network complexity. Species-level metagenomics revealed extensive reorganization within Pseudomonas, with contrasting shifts among multiple blotch-associated lineages, accompanied by changes in non-Pseudomonas taxa, including Mycetocola and Ewingella. Despite these taxonomic shifts, dominant Pseudomonas populations retained broadly conserved functional profiles, with disease-associated enrichment of pathways related to central metabolism, O-antigen biosynthesis, and peptidoglycan maturation. Casing communities underwent pronounced directional succession, shifting from early dominance by Exiguobacterium and Leuconostoc toward enrichment of Flavobacterium, Pedobacter, and Pseudomonas at later stages. Multiple blotch-associated Pseudomonas lineages were detected throughout casing development, while Pseudomonas increased from approximately 2% in early casing to 51% at harvest at the farm with higher disease incidence. Succession occurred without significant changes in alpha diversity, indicating that community development primarily reflected taxon replacement and redistribution. Conclusions: Our findings support a microbiome-centered framework for bacterial blotch in which disease is associated with host and stage dependent microbial succession, species-level community restructuring, and altered microbial connectivity. This framework extends beyond the single-pathogen paradigm and highlights bacterial blotch as a community-level disease process shaped by dynamic interactions between the mushroom host and its surrounding microbiome.

As a specialist herbivore, koalas rely on their gut microbiomes to help digest their toxic and fibrous diet of Eucalyptus leaves. Without these critical microbes, koalas may not be able to obtain the nutrients and energy they need to survive. As such, a large proportion of koalas that undergo rehabilitation for chlamydiosis develop gut dysbiosis from the antibiotic treatment and are either euthanized or die. Here we aimed to modify previously developed fresh faecal inoculation capsules for an extended shelf-life such that they could be applied in a clinical setting to prevent or treat gut dysbiosis. Using 16S rRNA gene amplicon sequencing we demonstrated that air-drying faecal material leads to an overgrowth of facultative anaerobes, whereas lyophilised material retains a similar microbial composition to fresh material. Initial survival of koala faecal microbes as assessed by live/dead staining combined with microscopy was high after lyophilisation regardless of which excipient was used, except for 20% glycerol that resulted in ~15% lower survival than other treatments. Lyophilised fine particles extracted from koala faeces and packaged into acid-resistant capsules maintained their original microbial composition and had high microbial survival over a year, regardless of the excipient used. Dry-fill, single-layer capsules maintained integrity after 10 hrs in synthetic koala stomach acid. These capsules may be useful in adapting the gut microbiomes of koalas to novel diets e.g. during translocations. However, attempts to apply the capsules in the clinical setting were unsuccessful due to unanticipated difficulties in administrating the capsules to sick koalas.

This study aims to evaluate the effectiveness of curcumin and penicillin-streptomycin as diluents on changes in the structure and diversity of the chicken semen microbiome during storage. Semen was collected from Kampung chickens (native to Indonesia) and divided into five treatment groups: diluted without antibiotics or curcumin (control), and diluted with additions of 10 micromolar, 20 micromolar, and 30 micromolar curcumin, and penicillin-streptomycin, respectively. The semen was stored at 5 C for 24 hours. The composition and diversity of the semen microbiome were analyzed using 16S full-length amplicon sequencing. Analysis of the top 10 species showed that Uncultured Saccharofermentans sp. and Porphyromonas somerae served as the most dominant and stable core microbiome across all treatments. Alpha diversity analysis showed that the addition of curcumin and penicillin-streptomycin reduced microbial richness (Observed, ChaO1, ACE, and Fisher) in a dose-dependent manner, yet maintained overall diversity (Shannon and Simpson), with the penicillin-streptomycin treatment resulting in the highest species evenness (InvSimpson). Beta diversity analysis revealed extreme separation of taxonomic abundance variance in the penicillin-streptomycin group, whereas the curcumin treatment exhibited a dose-dependent pattern of microbial abundance transition. Venn diagram analysis identified 415 OTUs as the core microbiome and confirmed that curcumin acts through selective filtering that stabilizes the ecosystem without triggering the proliferation of opportunistic taxa. Penicillin-streptomycin acts more rapidly and dominantly in suppressing/killing bacterial populations, however, the addition of curcumin is able to modulate the microbial ecosystem in a more balanced manner by suppressing the growth of harmful bacteria without compromising the integrity of the chicken semen environment.

Gelatinous zooplankton are abundant predators and prey in marine food webs, yet their role in helminth (parasitic worms) transmission remains poorly resolved. Here we combine a global synthesis of published records with new morphological and molecular observations from the Mediterranean, Red, Celtic, Baltic and North Seas to assess the ecological role of helminths associated with gelatinous zooplankton. We compiled 431 host-parasite association records from 89 sources, including 23 new records, and show that helminth occurrence and richness in gelatinous hosts are concentrated at temperate latitudes, contrasting with the classical latitudinal diversity gradient. Our sampling revealed markedly higher parasite prevalence, abundance and diversity in the Red Sea than in the Mediterranean, while no helminths were detected in gelatinous zooplankton sampled from the Baltic and North Seas. All helminths recorded were larval stages, indicating that gelatinous zooplankton function as key intermediate hosts in marine helminth life cycles. Molecular analyses identified cestodes, nematodes and digenean trematodes associated with cnidarians, ctenophores, and chaetognaths, including the first record of trematode larvae in a pelagic tunicate. Our findings challenge the view of gelatinous zooplankton as dead-end hosts and identify them as overlooked vectors that may shape marine parasite biogeography, food web connectivity and invasion dynamics.

Early-life microbial colonization is critical for shaping host development, yet how different maternal microbial reservoirs contribute to colonization of distinct offspring body sites remains poorly understood. Here, we explored microbial colonization and maturation of oral and rectal microbiota in puppies during the first postnatal week and assessed the contributions of maternal oral, vaginal, rectal, and milk microbiota to early colonization. We collected 505 samples from 33 dams and their litters at day 1 and day 8 postpartum and characterized microbial communities using 16S rRNA gene sequencing. We found that the pup oral microbiome on day 1 closely resembled maternal milk and vaginal microbiota. In contrast, the pup rectal microbiome was initially distinct from all maternal body sites, suggesting contributions from additional unmeasured sources. By day 8, both oral and rectal microbiomes exhibited signs of early maturation, with decreased relative abundance of opportunistic taxa and increased resemblance to maternal profiles. In dams, the vaginal microbiome showed the largest postpartum shift, while the rectal microbiome exhibited a smaller but significant change, and milk and oral microbiomes remained stable. Our results reveal that maternal contributions to early microbiome assembly are body-site specific, with oral and rectal microbiomes following distinct developmental trajectories during the first week of life, consistent with a dynamic colonization process shaped by both maternal and likely additional non-maternal sources. The observed patterns parallel those reported in humans, supporting the value of dogs as a comparative model for studying early-life microbiome colonization and maternal-offspring transmission in mammals.

Microbiomes alter host health and are shaped by transmission within and among host taxa. Microbes can alter host communities, yet whether the diversity of hosts reciprocally shapes microbial communities remains poorly resolved. Diverse host communities may promote microbiome diversity through elevated cross-species microbial spillover or reduce it by diluting conspecific transmission. Here, we manipulated the composition and diversity of pond zooplankton communities to test how host diversity and density alter host-associated and environmental microbiomes. Host diversity had opposing effects on microbiome alpha diversity for different zooplankton host species, increasing microbiome diversity in calanoid copepods while reducing it in Daphnia, with no effect on environmental microbes. Host diversity amplified cross-species microbial spillover for subordinate competitors (calanoid copepods) and diluted conspecific transmission for the dominant competitor (Daphnia). These contrasting responses reflected shifts in indicator microbes associated with each host taxon. Increasing host turnover generated higher microbiome turnover, demonstrating that microbiome assembly depends on host community composition. Our results indicate that contact with both conspecifics and heterospecifics alters animal microbiomes, and that host diversity can either promote the spillover or dilute the transmission of microbes in different zooplankton with potential effects on host health and ecosystems.

Multicellular animals emerged into a microbial world and continue to be influenced by sympatric bacterial species. Host-associated microbiomes are recognized as critical contributors to their living domiciles success. Detailing how microbiomes are assembled and maintained by their hosts has important implications for animal health, agricultural productivity, and biodiversity interventions across the tree of life. 10s-100s of bacterial genera are frequently detected in animal microbiomes, with some being uniquely and consistently detected in specific host species. Such discoveries have informed theories like phylosymbiosis that assert that host-associated microbiomes often reflect host species relatedness. We explored the predictive limits of this theory by performing a multi-scale dissimilarity-based comparative analyses using publicly-available microbiomes drawn from 113 insect species spanning 14 insect orders. We detected a moderate inter-order phylosymbiotic signal (Mantel test: r = 0.392, p<0.001) that was weakened (r = 0.173, p<0.001) when the Blattodea were excluded from the analysis. Intra-order level phylosymbiotic signals were detected for lepidopteran, hymenopteran, hemipteran, and blattodean host species microbiomes. These signals were often explained by obligate endosymbionts and/or nestmate-acquired gut bacteria, rather than by the total microbiomes. Host species-defining bacterial genera often represented a fraction of the total observed microbial diversity in their respective insect host species. We hypothesize that members of insect orders can exhibit considerable microbiome compositional variability where host speciation events may be minimally reflected in their microbiomes taxonomic membership, but host species-defining bacterial genera are likely when host behaviors or physiologies that facilitate reliable intra- and inter-generational bacterial transmission or acquisition are present.

Mucus is a critical interface between marine invertebrates and their environment and plays key roles in host defense. Despite its ecological importance, little is known about the microbial communities associated with mucus-producing corallimorpharians. Here, we used a multi-omics approach to characterize the mucus microbiome and immune repertoire of the Florida false coral, Ricordea florida. Culture-dependent and culture-independent analyses revealed that R. florida mucus harbors a bacterial community distinct from surrounding seawater and consistently dominated by Vibrio species. Whole-genome sequencing of 13 cultured Vibrio isolates representing nine distinct lineages revealed substantial taxonomic diversity, including several highly divergent strains that may represent undescribed species. Comparative genomic analyses identified enrichment of genes associated with carbohydrate acquisition, glycoside hydrolysis, and host colonization, including multiple components of the Tad/Flp adhesion system, suggesting adaptation to the mucus microenvironment. To investigate host factors that may shape microbial associations, we assembled and annotated a host transcriptome from healthy and immune-challenged polyps. This analysis identified a diverse innate immune repertoire, including Toll-like receptors, NOD-like receptors, lectins, scavenger receptors, complement-associated proteins, antiviral defense pathways, and membrane attack complex/perforin domain-containing effectors. Together, these findings demonstrate that R. florida supports a conserved, Vibrio-rich mucus microbiome and possesses a complex innate immune system capable of mediating host-microbe interactions at the mucosal surface. This study provides a foundation for understanding microbial colonization, immune defense, and holobiont function in an understudied cnidarian lineage.