Obligate Association Research Articles

Entomophthovirus: an insect-derived iflavirus that infects a behavior-manipulating fungal pathogen of dipterans.

We report a virus infecting Entomophthora muscae, a behavior-manipulating fungal pathogen of dipterans. The virus, which we name Berkeley Entomophthovirus, is a positive-strand RNA virus in the iflaviridae family of capsid-forming viruses, which are mostly known to infect insects. The viral RNA is expressed at high levels in fungal cells in vitro and during in vivo infections of Drosophila melanogaster, and virus particles can be seen intracellularly in E. muscae. This virus, of which we find two closely related variants in our culture of E. muscae, is also closely related to three different viruses reported from metagenomic surveys, two of which were isolated from wild dipterans, and a third isolated from wild ticks. By analyzing sequencing data from these earlier reports, we find abundant reads aligning to E. muscae specifically in the samples from which viral reads were sequenced. These data establish a wide and perhaps obligate association with E. muscae in the wild, consistent with our laboratory data that E. muscae is the host for these closely related viruses. Because of this, we propose the name Entomophthovirus (EV) for this group of highly related virus variants. As other members of the iflaviridae have been reported to cause behavioral changes in insects, we speculate on the possibility that EV plays a role in the behavioral manipulation of flies infected with E. muscae.

Discovery of a novel symbiotic lineage associated with a hematophagous leech from the genus Haementeria.

Obligate symbiotic associations with a nutritional base have likely evolved more than once in strict blood-feeding leeches. Unlike those symbioses found in hematophagous arthropods, the nature, identity, and evolutionary history of these remains poorly studied. In this work, we further explored obligate nutritional associations between Haementeria leeches and their microbial symbionts, which led to the unexpected discovery of a novel symbiosis with a member of the Pluralibacter genus. When compared to Providencia siddallii, an obligate nutritional symbiont of other Haementeria leeches, this novel bacterial symbiont shows convergent retention of the metabolic pathways involved in B vitamin biosynthesis. Moreover, the genomic characteristics of this Pluralibacter symbiont suggest a more recent association than that of Pr. siddallii and Haementeria. We conclude that the once-thought stable associations between blood-feeding Glossiphoniidae and their symbionts (i.e., one bacteriome structure, one symbiont lineage) can break down, mirroring symbiont turnover observed in various arthropod lineages.

Importance of complex sponges as habitat and feeding substrata for coral reef fishes

Coral reef fishes are usually assumed to be most strongly associated with reef-building corals. However, sponges can be a significant structural component of coral reef ecosystems and their framework can enhance the local abundance and biodiversity of fish assemblages. Little is known regarding the range of fish species using complex sponges as either shelter or feeding substrata. Here we use a combination of stationary video cameras and focal animal sampling to document fish species positively associated with complex sponges in Kimbe Bay, Papua New Guinea. Stationary cameras identified 45 fish species using the sponges for either shelter, feeding substrata or as sites for ambush predation. A guild of 10 individual fish species from five families (Blenniidae, Chaetodontidae, Gobiidae, Labridae and Pomacentridae) were observed to quantify sponge and other habitat use and compared with habitat availability to determine the level of sponge selectivity. One species, Pleurosicya elongata (the Slender Spongegoby), lived in obligate association with Ianthella basta (Elephant Ear sponge), and there was a positive relationship between sponge size and number of resident fish, however this was not significant for all life stages. Five other fish species appeared to preferentially select sponges as habitat (Amblyglyphidodon aureus, Chaetodon kleinii, Coradion chrysozonus, Escenius prooculis and Pomacentrus nigromanus), while for others, sponge use appeared incidental. When selectivity indices were calculated for specific sponge species it was apparent that some fishes exhibited preferences for particular sponge species or growth forms. These results suggest more fish species may be reliant on sponges than is widely appreciated.

Green and red fluorescent strains of Xenorhabdus griffiniae HGB2511, the bacterial symbiont of the nematode Steinernema hermaphroditum (India).

Steinernema entomopathogenic nematodes form specific, obligate symbiotic associations with gram-negative, gammaproteobacteria members of the Xenorhabdus genus. Together, the nematodes and symbiotic bacteria infect and kill insects, utilize the nutrient-rich cadaver for reproduction, and then reassociate, the bacteria colonizing the nematodes' anterior intestines before the nematodes leave the cadaver to search for new prey. In addition to their use in biocontrol of insect pests, these nematode-bacteria pairs are highly tractable experimental laboratory models for animal-microbe symbiosis and parasitism research. One advantageous feature of entomopathogenic nematode model systems is that the nematodes are optically transparent, which facilitates direct observation of nematode-associated bacteria throughout the lifecycle. In this work, green- and red-fluorescently labeled X. griffiniae HGB2511 bacteria were created and associated with their S . hermaphroditum symbiotic nematode partners and observed using fluorescence microscopy. As expected, the fluorescent bacteria were visible as a colonizing cluster in the lumen of the anterior intestinal caecum of the infective stage of the nematode. These tools allow detailed observations of X. griffiniae localization and interactions with its nematode and insect host tissues throughout their lifecycles.

Novel evolutionary insights on the interactions of the Holosporales (Alphaproteobacteria) with eukaryotic hosts from comparative genomics.

Holosporales are an alphaproteobacterial order engaging in obligate and complex associations with eukaryotes, in particular protists. The functional and evolutionary features of those interactions are still largely undisclosed. Here, we sequenced the genomes of two members of the species Bealeia paramacronuclearis (Holosporales, Holosporaceae) intracellularly associated with the ciliate protist Paramecium, which resulted in high correspondence. Consistent with the short-branched early-divergent phylogenetic position, Bealeia presents a larger functional repertoire than other Holosporaceae, comparable to those of other Holosporales families, particularly for energy metabolism and motility. Our analyses indicate that different Holosporales likely experienced at least partly autonomous genome reduction and adaptation to host interactions, for example regarding dependence on host biotin driven by multiple independent horizontal acquisitions of transporters. Among Alphaproteobacteria, this is reminiscent of the convergently evolved Rickettsiales, which however appear more diverse, possibly due to a probably more ancient origin. We identified in Bealeia and other Holosporales the plasmid-encoded putative genetic determinants of R-bodies, which may be involved in a killer trait towards symbiont-free hosts. While it is not clear whether these genes are ancestral or recently horizontally acquired, an intriguing and peculiar role of R-bodies is suggested in the evolution of the interactions of multiple Holosporales with their hosts.

A theoretical model for host-controlled regulation of symbiont density.

There is growing empirical evidence that animal hosts actively control the density of their mutualistic symbionts according to their requirements. Such active regulation can be facilitated by compartmentalization of symbionts within host tissues, which confers a high degree of control of the symbiosis to the host. Here, we build a general theoretical framework to predict the underlying ecological drivers and evolutionary consequences of host-controlled endosymbiont density regulation for a mutually obligate association between a host and a compartmentalized, vertically transmitted symbiont. Building on the assumption that the costs and benefits of hosting a symbiont population increase with symbiont density, we use state-dependent dynamic programming to determine an optimal strategy for the host, i.e., that which maximizes host fitness, when regulating the density of symbionts. Simulations of active host-controlled regulation governed by the optimal strategy predict that the density of the symbiont should converge to a constant level during host development, and following perturbation. However, a similar trend also emerges from alternative strategies of symbiont regulation. The strategy which maximizes host fitness also promotes symbiont fitness compared to alternative strategies, suggesting that active host-controlled regulation of symbiont density could be adaptive for the symbiont as well as the host. Adaptation of the framework allowed the dynamics of symbiont density to be predicted for other host-symbiont ecologies, such as for non-essential symbionts, demonstrating the versatility of this modelling approach.

“Candidatus Intestinibacterium parameciiphilum”—member of the “Candidatus Paracaedibacteraceae” family (Alphaproteobacteria, Holosporales) inhabiting the ciliated protist Paramecium

Protists frequently host diverse bacterial symbionts, in particular those affiliated with the order Holosporales (Alphaproteobacteria). All characterised members of this bacterial lineage have been retrieved in obligate association with a wide range of eukaryotes, especially multiple protist lineages (e.g. amoebozoans, ciliates, cercozoans, euglenids, and nucleariids), as well as some metazoans (especially arthropods and related ecdysozoans). While the genus Paramecium and other ciliates have been deeply investigated for the presence of symbionts, known members of the family “Candidatus Paracaedibacteraceae” (Holosporales) are currently underrepresented in such hosts. Herein, we report the description of “Candidatus Intestinibacterium parameciiphilum” within the family “Candidatus Paracaedibacteraceae”, inhabiting the cytoplasm of Paramecium biaurelia. This novel bacterium is almost twice as big as its relative “Candidatus Intestinibacterium nucleariae” from the opisthokont Nuclearia and does not present a surrounding halo. Based on phylogenetic analyses of 16S rRNA gene sequences, we identified six further potential species-level lineages within the genus. Based on the provenance of the respective samples, we investigated the environmental distribution of the representatives of “Candidatus Intestinibacterium” species. Obtained results are consistent with an obligate endosymbiotic lifestyle, with protists, in particular freshwater ones, as hosts. Thus, available data suggest that association with freshwater protists could be the ancestral condition for the members of the “Candidatus Intestinibacterium” genus.

Genomic characterization of a novel, widely distributed Mycoplasma species "Candidatus Mycoplasma mahonii" associated with the brittlestar Gorgonocephalus chilensis.

Symbiotic relationships are ubiquitous throughout the world's oceans, yet for many marine organisms, including those in the high latitudes, little is understood about symbiotic associations and functional relationships. From a recently determined genome sequence of a filter-feeding basket star from Argentina, Gorgonocephalus chilensis, we discovered a novel Mycoplasma species with a 796Kb genome (CheckM completeness of 97.9%, G+C content = 30.1%). Similar to other Mycoplasma spp. within Mycoplasmatota, genomic analysis of the novel organism revealed reduced metabolic pathways including incomplete biosynthetic pathways, suggesting an obligate association with their basket star host. Results of 16S rRNA and multi-locus phylogenetic analyses revealed that this organism belonged to a recently characterized non-free-living lineage of Mycoplasma spp. specifically associated with marine invertebrate animals. Thus, the name "Candidatus Mycoplasma mahonii" is proposed for this novel species. Based on 16S rRNA PCR-screening, we found that Ca. M. mahonii also occurs in Gorgonocephalus eucnemis from the Northwest Pacific and other Gorgonocephalus chilensis from Argentinian waters. The level of sequence conservation within Ca. M. mahonii is considerable between widely disparate high-latitude Gorgonocephalus species, suggesting that oceanic dispersal of this microbe may be greater than excepted.

How to be a fig nematode

Fig (Moraceae: Ficus) species host vast communities of organisms that are bound together by complicated ecological networks that have influenced community structure and dynamics over evolutionary timescales. Much attention has been paid to the mutualism between figs and their specialized pollinating fig wasps, as well as with often antagonistic non-pollinating fig wasps. Equally ubiquitous to fig systems, but much less understood are the multitude of nematode groups that have independently evolved obligate associations with pollinating fig wasps and proliferate inside fig syconia. In this review we describe what is currently known (and unknown) about these numerous and increasingly studied nematode taxa and how they interact with the fig systems they inhabit. We identify the groups that are currently understood to associate with fig pollinators and outline their known species distributions and evolutionary history, where possible. Special attention is paid to the life history of these nematode groups, especially which features of nematode biology are generalizable across groups and what idiosyncratic peculiarities exist within individual genera. We outline key biological features including host choice, dispersal, disembarkation, diet, mating, and proliferation within figs. We address biological conundrums that have been raised following observational work such as, why do nematodes sometimes infect the wrong host? What adaptations were required for nematodes to successfully adapt and coexist with figs and their pollinators for millions of years? How do nematodes overcome the constraints of low mating group size? Finally, we outline key considerations, gaps in the knowledge, and limitations to expand this field forward towards promising areas of future research. Through better understanding of fig nematodes, we stand to not only know more about Ficus communities, but also more about the evolution and maintenance of interspecific interactions, development, adaptation, and co-evolution in general.

Breeding biology of the African Pygmy Falcon: long-term variation and seasonal decline in breeding performance of an arid zone raptor

Birds of prey are apex predators and understanding their life history can serve as a valuable baseline for investigating their ecological role. Pygmy Falcons (Polihieraxsemitorquatus), Africa’s smallest diurnal raptor, have evolved to be obligate associates of Sociable Weaver (Philetairussocius) colonies throughout their southern African range. As a predator, Pygmy Falcons likely impact prey communities in this system including their host and other colony associates. However, no study has explored their breeding biology in detail using long-term data. We provide results from 10 years of data collected between 2011 and 2020. We followed 66 unique Pygmy Falcon territories and 323 breeding attempts in the Kalahari, South Africa. We explored annual variation in the population density of Pygmy Falcons and investigated between- and within-season variation in reproductive investment and output. The highest average density was recorded in 2015 and the lowest in 2019 and 2020. Our results show that the breeding occurs between early austral spring (August) and late summer (February), with a peak from September to December. The maximum and most frequent clutch size was three eggs and falcons usually initiated only one breeding attempt (but up to three) in a season. The incubation and nestling periods lasted on average 33 (±SD 4) and 37 (± 5) days, respectively. A three-egg clutch size and number of fledglings produced varied between seasons, and breeding success showed a seasonal decline across the breeding season. Falcons were 7 times less likely to lay a 3-egg clutch in 2019 than in 2011 and 2012, and breeding attempts initiated in September were 3.4 times more likely to be successful than those initiated in November. Predation accounted for 49.5% of breeding failures. Our results show significant variation in the breeding parameters of these falcons, but further studies are needed to understand what drives these variations.

Trophobiosis between a new species of Acropyga (Hymenoptera, Formicidae) and new Neochavesia (Hemiptera, Xenococcidae) from Peru, and establishment of the Acropygasmithii species-group.

We describe a new pair of trophobiotic partners from the ant genus Acropyga and the root mealybug genus Neochavesia. A recent field study on Acropyga ants and associated root mealybugs, conducted in the Peruvian Amazon, led to the discovery of Acropygamanuense LaPolla & Schneider, sp. nov. and its root mealybug symbiont Neochavesiapodexuta Schneider & LaPolla, sp. nov. The new root mealybug belongs to the family Xenococcidae, whose members are all obligate associates of Acropyga ants. Providing joint descriptions of new mutualist partners in the same article is a novel approach for this system, and it offers benefits to the ongoing study of mutualism and patterns of association among these symbiotic ants and scales. Here, we also begin to revise the species-group composition of Acropyga by establishing the smithii species-group, and we provide updated information to aid in identifying the new ant species and root mealybug species.

Prokaryotic and eukaryotic microbial communities associated with coral species have high host specificity in the South China Sea

Reef-building corals are well known for their obligate association with Symbiodiniaceae, and an array of other microbes, including bacteria, fungi, and symbiotic algae (i.e., total microbiome), which together form the coral holobiont. The total microbiome plays an intricate part in maintaining the homeostasis of the coral holobiont and is closely associated with host health. However, the composition of the coral associated microbiome and interaction between its different members remains elusive because few analyses have bridged taxonomically disparate groups. This research gaps have prevented a holistic understanding of the total microbiome. Thus, to simultaneously characterize the bacterial, fungal and symbiotic algal communities associated with different coral species, and explore the relationship between these symbionts and coral health, healthy and bleached tissues from four coral species, Acropora muricata, Galaxea fascicularis, Platygyra daedalea, and Pavona explanulata, were collected from the Xisha Islands of the South China Sea. Using high throughput sequencing, a high degree of host-specificity was observed among bacterial, fungal, and algal groups across coral species. There were no obvious changes in the microbial community structure of apparently healthy and bleached corals, but host bleaching allowed colonization of the holobionts by diverse opportunistic microbes, resulting in a significant elevation in the α-diversity of microbial communities. In addition, co-occurrence analysis of the coral microbiota also identified more complex microbial interactions in bleached corals than in healthy ones. In summary, this study characterized the structure of coral-associated microbiomes across four coral species, and systematically studied microbiome differences between healthy and bleached corals. The findings improve our understanding of the heterogeneity of symbiotic microorganisms and the impact of coral's physiological status on its associated microbial communities composition.

To explore strange new worlds – The diversification in Tremella caloplacae was linked to the adaptive radiation of the Teloschistaceae

Lichenicolous fungi are a heterogeneous group of organisms that grow exclusively on lichens, forming obligate associations with them. It has often been assumed that cospeciation has occurred between lichens and lichenicolous fungi, but this has been seldom analysed from a macroevolutionary perspective. Many lichenicolous species are rare or are rarely observed, which results in frequent and large gaps in the knowledge of the diversity of many groups. This, in turn, hampers evolutionary studies that necessarily are based on a reasonable knowledge of this diversity. Tremella caloplacae is a heterobasidiomycete growing on various hosts from the lichen-forming family Teloschistaceae, and evidence suggests that it may represent a species complex. We combine an exhaustive sampling with molecular and ecological data to study species delimitation, cophylogenetic events and temporal concordance of this association. Tremella caloplacae is here shown to include at least six distinct host-specific lineages (=putative species). Host switch is the dominant and most plausible event influencing diversification and explaining the coupled evolutionary history in this system, although cospeciation cannot be discarded. Speciation in T. caloplacae would therefore have occurred coinciding with the rapid diversification – by an adaptive radiation starting in the late Cretaceous – of their hosts. New species in T. caloplacae would have developed as a result of specialization on diversifying lichen hosts that suddenly offered abundant new ecological niches to explore or adapt to.

Octocorals in the Gulf of Aqaba exhibit high photosymbiont fidelity.

Symbiotic associations, widespread in terrestrial and marine ecosystems, are of considerable ecological importance. Many tropical coral species are holobionts, formed by the obligate association between a cnidarian host and endosymbiotic dinoflagellates of the family Symbiodiniaceae. The latter are abundant on coral reefs from very shallow water down to the upper mesophotic zone (30-70 m). The research on scleractinians has revealed that the photosymbiont lineages present in the cnidarian host play an important role in the coral's ability to thrive under different environmental conditions, such as light regime and temperature. However, little is known regarding octocoral photosymbionts, and in particular regarding those found deeper than 30 m. Here, we used ribosomal (ITS2) and chloroplast (23S) markers to uncover, for the first time, the dominant Symbiodiniaceae taxa present in 19 mesophotic octocoral species (30-70 m depth) from the Gulf of Aqaba/Eilat (northern Red Sea). In addition, using high-throughput sequencing of the ITS2 region we characterized both the dominant and the rare Symbiodiniaceae lineages found in several species across depth. The phylogenetic analyses of both markers were in agreement and revealed that most of the studied mesophotic octocorals host the genus Cladocopium. Litophyton spp. and Klyxum utinomii were exceptions, as they harbored Symbiodinium and Durusdinium photosymbionts, respectively. While the dominant algal lineage of each coral species did not vary across depth, the endosymbiont community structure significantly differed between host species, as well as between different depths for some host species. The findings from this study contribute to the growing global-catalogue of Cnidaria-Symbiodiniaceae associations. Unravelling the Symbiodiniaceae composition in octocoral holobionts across environmental gradients, depth in particular, may enable a better understanding of how specialized those associations are, and to what extent coral holobionts are able to modify their photosymbionts.

The role of the length spines of echinoids in their association with crustacean and fish in Los Cabos, Baja California Sur, Mexico

Introduction: Echinoids (sea urchins) provide shelter for a variety of facultative or obligatory ectosymbionts. Objective: To evaluate the hypothesis that decapods and fishes prefer to associate with echinoid individuals and species that have longer spines. Methods: We visually studied the frequency of decapod crustaceans and fishes associated with echinoids in shallow water (< 4 m) and deeper water (5-20 m) at Los Cabos, Baja California Sur, Mexico, during 1-6 January 2019. Results: We inspected 1 058 echinoids of six species. Five decapod species associated with three species of echinoids. When compared with other echinoid species, in shallow water, decapods associated 5.1 times more often with the longest-spined echinoid Diadema mexicanum (7.0 times more decapods per individual D. mexicanum); in deeper water, association frequency was similar for all echinoid species. Fourteen fish species associated with four echinoid species. In shallow water, fishes associated 2.6 times more with D. mexicanum (4.5 times more fishes per individual). There was no preferred echinoid species in deeper water. Longer-spined D. mexicanum had more decapods and fishes. Associations were more frequent in shallow water. Multiple individuals and species of decapods and fish often associated together with a single D. mexicanum. The decapod that presumably is Tuleariocaris holthuisi showed a possible obligatory association with one of the equinoids (D. mexicanum); the other decapods and all fish species are facultative associates. Conclusion: Our results support the hypothesis that decapods and fishes associate most frequently with echinoid with the longest spines, presumably to reduce the risk of predation.

Functional differentiation determines the molecular basis of the symbiotic lifestyle of Ca. Nanohaloarchaeota

BackgroundCandidatus Nanohaloarchaeota, an archaeal phylum within the DPANN superphylum, is characterized by limited metabolic capabilities and limited phylogenetic diversity and until recently has been considered to exclusively inhabit hypersaline environments due to an obligate association with Halobacteria. Aside from hypersaline environments, Ca. Nanohaloarchaeota can also have been discovered from deep-subsurface marine sediments.ResultsThree metagenome-assembled genomes (MAGs) representing a new order within the Ca. Nanohaloarchaeota were reconstructed from a stratified salt crust and proposed to represent a novel order, Nucleotidisoterales. Genomic features reveal them to be anaerobes capable of catabolizing nucleotides by coupling nucleotide salvage pathways with lower glycolysis to yield free energy. Comparative genomics demonstrated that these and other Ca. Nanohaloarchaeota inhabiting saline habitats use a “salt-in” strategy to maintain osmotic pressure based on the high proportion of acidic amino acids. In contrast, previously described Ca. Nanohaloarchaeota MAGs from geothermal environments were enriched with basic amino acids to counter heat stress. Evolutionary history reconstruction revealed that functional differentiation of energy conservation strategies drove diversification within Ca. Nanohaloarchaeota, further leading to shifts in the catabolic strategy from nucleotide degradation within deeper lineages to polysaccharide degradation within shallow lineages.ConclusionsThis study provides deeper insight into the ecological functions and evolution of the expanded phylum Ca. Nanohaloarchaeota and further advances our understanding on the functional and genetic associations between potential symbionts and hosts.6R3_PbsEzFSPy16NiyaaD4Video

Analysis of Risk Factors for Vulnerable Plaque Formation and Pathogenic in Carotid Artery.

The formation of vulnerable carotid artery plaque may be closely related to a single factor or caused by multiple factors. This paper discusses the pathogenic risk factors for vulnerable plaque in patients with severe internal carotid artery (ICA) stenosis who received endarterectomy through regression analysis. A total of 98 patients with a complete clinical and laboratory assessment underwent carotid endarterectomy. Metabolic syndrome (MetS) and MetS components, ICA plaque thickness and ICA peak systolic velocity, previous ischemic stroke or transient ischemic attack (TIA), and other risk factors were included in the pathogenic risk factor for vulnerable plaque. Univariate logistic regression analysis was used to determine vulnerable carotid plaque risk factors. If P <0.2, it was considered potential confounders. Binary logistic regression model was controlled for potential confounders. Among the 98 patients, stable carotid plaques 38 (39%) and unstable carotid plaques 60 (61%), male 76 (77.6%) and female 22 (22.4%), and Han Chinese 68 (68.4%) and Mongols 30 (30.6%). Univariate logistic regression to P <0.2 has 6 risk factors, which are previous ischemic stroke or TIA, ICA peak systolic velocity, ICA plaque thickness, body mass index, total cholesterol, and alcohol consumption. The significant result of the binary logistic regression analysis was the previous ischemic stroke or TIA (OR=4.52; 95% CI, 1.67-12.09), P =0.003 and ICA peak systolic velocity (OR=1.01; 95% CI, 1.00-1.02), P =0.014. The patients with previous ischemic stroke or TIA and higher ICA peak systolic velocity are associated with vulnerable plaque pathogenic features. There is no obligatory association between MetS and formation of carotid plaque vulnerability.

The Genomic Basis of Evolutionary Novelties in a Leafhopper.

Evolutionary innovations generate phenotypic and species diversity. Elucidating the genomic processes underlying such innovations is central to understanding biodiversity. In this study, we addressed the genomic basis of evolutionary novelties in the glassy-winged sharpshooter (Homalodisca vitripennis, GWSS), an agricultural pest. Prominent evolutionary innovations in leafhoppers include brochosomes, proteinaceous structures that are excreted and used to coat the body, and obligate symbiotic associations with two bacterial types that reside within cytoplasm of distinctive cell types. Using PacBio long-read sequencing and Dovetail Omni-C technology, we generated a chromosome-level genome assembly for the GWSS and then validated the assembly using flow cytometry and karyotyping. Additional transcriptomic and proteomic data were used to identify novel genes that underlie brochosome production. We found that brochosome-associated genes include novel gene families that have diversified through tandem duplications. We also identified the locations of genes involved in interactions with bacterial symbionts. Ancestors of the GWSS acquired bacterial genes through horizontal gene transfer (HGT), and these genes appear to contribute to symbiont support. Using a phylogenomics approach, we inferred HGT sources and timing. We found that some HGT events date to the common ancestor of the hemipteran suborder Auchenorrhyncha, representing some of the oldest known examples of HGT in animals. Overall, we show that evolutionary novelties in leafhoppers are generated by the combination of acquiring novel genes, produced both de novo and through tandem duplication, acquiring new symbiotic associations that enable use of novel diets and niches, and recruiting foreign genes to support symbionts and enhance herbivory.

First Neotropical record of the association between brown sclerotium-forming fungi and termite eggs in a nest of Coptotermes gestroi (Blattaria, Isoptera, Rhinotermitidae)

Insects and fungi are abundant in many environments, in which facultative and/or obligate associations involving these groups have been established during evolution. In termites, mutualism with fungi is well reported for some termite lineages (e.g., Macrotermitinae). Within some subterranean termite species (Rhinotermitidae), egg-mimicking fungi, also referred to as "termite "balls", are often harbored inside the nest, mixed to the egg piles. Such interaction seems to be advantageous for both partners since the fungi are protected inside the nest while they may serve as an additional food source and also provide cellulases which may be incorporated into the termite digestive process. Although such mutualism has been reported for seven species of Reticulitermes and Coptotermes formosanus, all the samplings were restricted to temperate regions. Here, we provide the first Neotropical record of this termite-fungus association, and the first report for Coptotermes gestroi. The morphological characters of the "termite balls" observed in a C. gestroi nest resemble those already reported for Reticulitermes spp. and the congeneric species C. formosanus. They include a color ranging from light to dark brown, spherical shape, and a reduced diameter (0.23-0.34mm). Our findings provide new insights into the geographical distribution of the association between termites and sclerotium-forming fungi. Future genetic analyses will be valuable aiming to identify the egg-mimicking fungi associated with C. gestroi and shed light on the evolution of this fascinating symbiosis.

The microbiome of a bacterivorous marine choanoflagellate contains a resource-demanding obligate bacterial associate

Microbial predators such as choanoflagellates are key players in ocean food webs. Choanoflagellates, which are the closest unicellular relatives of animals, consume bacteria and also exhibit marked biological transitions triggered by bacterial compounds, yet their native microbiomes remain uncharacterized. Here we report the discovery of a ubiquitous, uncultured bacterial lineage we name Candidatus Comchoanobacterales ord. nov., related to the human pathogen Coxiella and physically associated with the uncultured marine choanoflagellate Bicosta minor. We analyse complete ‘Comchoano’ genomes acquired after sorting single Bicosta cells, finding signatures of obligate host-dependence, including reduction of pathways encoding glycolysis, membrane components, amino acids and B-vitamins. Comchoano encode the necessary apparatus to import energy and other compounds from the host, proteins for host-cell associations and a type IV secretion system closest to Coxiella’s that is expressed in Pacific Ocean metatranscriptomes. Interactions between choanoflagellates and their microbiota could reshape the direction of energy and resource flow attributed to microbial predators, adding complexity and nuance to marine food webs.