Arthropod Gut Research Articles

Genome reduction in novel, obligately methyl-reducing Methanosarcinales isolated from arthropod guts (Methanolapillus gen. nov. and Methanimicrococcus).

Recent metagenomic studies have identified numerous lineages of hydrogen-dependent, obligately methyl-reducing methanogens. Yet, only a few representatives have been isolated in pure culture. Here, we describe six new species with this capability in the family Methanosarcinaceae (order Methanosarcinales), which makes up a substantial fraction of the methanogenic community in arthropod guts. Phylogenomic analysis placed the isolates from cockroach hindguts into the genus Methanimicrococcus (M. hacksteinii, M. hongohii, and M. stummii) and the isolates from millipede hindguts into a new genus, Methanolapillus (M. africanus, M. millepedarum, and M. ohkumae). Members of this intestinal clade, which includes also uncultured representatives from termites and vertebrates, have substantially smaller genomes (1.6-2.2 Mbp) than other Methanosarcinales. Genome reduction was accompanied by the loss of the upper part of the Wood-Ljungdahl pathway, several energy-converting membrane complexes (Fpo, Ech, and Rnf), and various biosynthetic pathways. However, genes involved in the protection against reactive oxygen species (catalase and superoxide reductase) were conserved in all genomes, including cytochrome bd (CydAB), a high-affinity terminal oxidase that may confer the capacity for microaerobic respiration. Since host-associated Methanosarcinales are nested within omnivorous lineages, we conclude that the specialization on methyl groups is an adaptation to the intestinal environment.

Mechanism of complement inhibition by a mosquito protein revealed through cryo-EM

Salivary complement inhibitors occur in many of the blood feeding arthropod species responsible for transmission of pathogens. During feeding, these inhibitors prevent the production of proinflammatory anaphylatoxins, which may interfere with feeding, and limit formation of the membrane attack complex which could damage arthropod gut tissues. Salivary inhibitors are, in many cases, novel proteins which may be pharmaceutically useful or display unusual mechanisms that could be exploited pharmaceutically. Albicin is a potent inhibitor of the alternative pathway of complement from the saliva of the malaria transmitting mosquito, Anopheles albimanus. Here we describe the cryo-EM structure of albicin bound to C3bBb, the alternative C3 convertase, a proteolytic complex that is responsible for cleavage of C3 and amplification of the complement response. Albicin is shown to induce dimerization of C3bBb, in a manner similar to the bacterial inhibitor SCIN, to form an inactive complex unable to bind the substrate C3. Size exclusion chromatography and structures determined after 30 minutes of incubation of C3b, factor B (FB), factor D (FD) and albicin indicate that FBb dissociates from the inhibited dimeric complex leaving a C3b-albicin dimeric complex which apparently decays more slowly.

Global diversity of enterococci and description of 18 previously unknown species

Enterococci are gut microbes of most land animals. Likely appearing first in the guts of arthropods as they moved onto land, they diversified over hundreds of millions of years adapting to evolving hosts and host diets. Over 60 enterococcal species are now known. Two species, Enterococcus faecalis and Enterococcus faecium, are common constituents of the human microbiome. They are also now leading causes of multidrug-resistant hospital-associated infection. The basis for host association of enterococcal species is unknown. To begin identifying traits that drive host association, we collected 886 enterococcal strains from widely diverse hosts, ecologies, and geographies. This identified 18 previously undescribed species expanding genus diversity by >25%. These species harbor diverse genes including toxins and systems for detoxification and resource acquisition. Enterococcus faecalis and E. faecium were isolated from diverse hosts highlighting their generalist properties. Most other species showed a more restricted distribution indicative of specialized host association. The expanded species diversity permitted the Enterococcus genus phylogeny to be viewed with unprecedented resolution, allowing features to be identified that distinguish its four deeply rooted clades, and the entry of genes associated with range expansion such as B-vitamin biosynthesis and flagellar motility to be mapped to the phylogeny. This work provides an unprecedentedly broad and deep view of the genus Enterococcus, including insights into its evolution, potential new threats to human health, and where substantial additional enterococcal diversity is likely to be found.

Inhibition of vertebrate complement system by hematophagous arthropods: inhibitory molecules, mechanisms, physiological roles, and applications.

In arthropods, hematophagy has arisen several times throughout evolution. This specialized feeding behavior offered a highly nutritious diet obtained during blood feeds. On the other hand, blood-sucking arthropods must overcome problems brought on by blood intake and digestion. Host blood complement acts on the bite site and is still active after ingestion, so complement activation is a potential threat to the host's skin feeding environment and to the arthropod gut enterocytes. During evolution, blood-sucking arthropods have selected, either in their saliva or gut, anticomplement molecules that inactivate host blood complement. This review presents an overview of the complement system and discusses the arthropod's salivary and gut anticomplement molecules studied to date, exploring their mechanism of action and other aspects related to the arthropod-host-pathogen interface. The possible therapeutic applications of arthropod's anticomplement molecules are also discussed.

PcEiger links the Imd/Relish pathway to ROSproduction in the intestine of the red swampcrayfish.

In the arthropod gut, commensal microbiota maintain the immune deficiency (Imd)/Relish pathway for expression of antimicrobial peptides, whereas pathogenic bacteria induce dual oxidase 2 (Duox2) for production of extracellular microbicidal reactive oxygen species (ROS). The Imd/Relish pathway and the Duox2/ROS system are regarded as independent systems. Here, we report that these two systems are bridged by the tumor necrosis factor (TNF) ortholog PcEiger in the red swamp crayfish Procambarus clarkii. PcEiger expression is induced by commensal bacteria or the Imd/Relish pathway. PcEiger knockdown alters bacterial abundance and community composition due to variations in the oxidative status of the intestine. PcEiger induces Duox2 expression and ROS production by regulating the activity of the transcription factor Atf2. Moreover, PcEiger mediates regulation of the Duox2/ROS system by commensal bacteria and the Imd/Relish pathway. Our findings suggest that the Imd/Relish pathway regulates the Duox2/ROS system via PcEiger in P. clarkii, and they provide insights into the crosstalk between these two important mechanisms for arthropod intestinal immunity.

Mycoparasites, Gut Dwellers, and Saprotrophs: Phylogenomic Reconstructions and Comparative Analyses of Kickxellomycotina Fungi.

Improved sequencing technologies have profoundly altered global views of fungal diversity and evolution. High-throughput sequencing methods are critical for studying fungi due to the cryptic, symbiotic nature of many species, particularly those that are difficult to culture. However, the low coverage genome sequencing (LCGS) approach to phylogenomic inference has not been widely applied to fungi. Here we analyzed 171 Kickxellomycotina fungi using LCGS methods to obtain hundreds of marker genes for robust phylogenomic reconstruction. Additionally, we mined our LCGS data for a set of nine rDNA and protein coding genes to enable analyses across species for which no LCGS data were obtained. The main goals of this study were to: 1) evaluate the quality and utility of LCGS data for both phylogenetic reconstruction and functional annotation, 2) test relationships among clades of Kickxellomycotina, and 3) perform comparative functional analyses between clades to gain insight into putative trophic modes. In opposition to previous studies, our nine-gene analyses support two clades of arthropod gut dwelling species and suggest a possible single evolutionary event leading to this symbiotic lifestyle. Furthermore, we resolve the mycoparasitic Dimargaritales as the earliest diverging clade in the subphylum and find four major clades of Coemansia species. Finally, functional analyses illustrate clear variation in predicted carbohydrate active enzymes and secondary metabolites (SM) based on ecology, that is biotroph versus saprotroph. Saprotrophic Kickxellales broadly lack many known pectinase families compared with saprotrophic Mucoromycota and are depauperate for SM but have similar numbers of predicted chitinases as mycoparasitic.

DNA High-Throughput Sequencing for Arthropod Gut Content Analysis to Evaluate Effectiveness and Safety of Biological Control Agents.

The search for effective biological control agents without harmful non-target effects has been constrained by the use of impractical (field direct observation) or imprecise (cage experiments) methods. While advances in the DNA sequencing methods, more specifically the development of high-throughput sequencing (HTS), have been quickly incorporated in biodiversity surveys, they have been slow to be adopted to determine arthropod prey range, predation rate and food web structure, and critical information to evaluate the effectiveness and safety of a biological control agent candidate. The lack of knowledge on how HTS methods could be applied by ecological entomologists constitutes part of the problem, although the lack of expertise and the high cost of the analysis also are important limiting factors. In this review, we describe how the latest HTS methods of metabarcoding and Lazaro, a method to identify prey by mapping unassembled shotgun reads, can serve biological control research, showing both their power and limitations. We explain how they work to determine prey range and also how their data can be used to estimate predation rates and subsequently be translated into food webs of natural enemy and prey populations helping to elucidate their role in the community. We present a brief history of prey detection through molecular gut content analysis and also the attempts to develop a more precise formula to estimate predation rates, a problem that still remains. We focused on arthropods in agricultural ecosystems, but most of what is covered here can be applied to natural systems and non-arthropod biological control candidates as well.

Trichomycetes from the Columbia Mountains, British Columbia, Canada

Trichomycetes are a group of fungi and protists living in the guts of immature aquatic insects and other arthropods. The nature of this relationship is poorly understood but it is mostly described as obligately commensalistic, with the microbes exploiting a unique microhabitat. Collections of trichomycetes were made from freshwater insects in aquatic habitats near Castlegar in the Columbia Mountains, part of the Rocky Mountain Range in southeastern British Columbia. The collection sites were at altitudes ranging from 435 to 935 m and were made in July 2006. A total of 32 species were recorded including four new species, Harpella filispora sp. nov., Orphella columbiensis sp. nov., Smittium nodiradicatum sp. nov., and Smitium basiangustatum sp. nov. described here. These are the first records from Western Canada and two, Austrosmittium patagonicum and Glotzia distorta are new continental records. These findings are compared to reports from the western edge of the Rocky Mountain range near the Boise Mountains in Idaho and the Southern Rockies in Colorado.

Gut Microbiota of Ostrinia nubilalis Larvae Degrade Maize Cellulose.

Most arthropod guts harbor diverse microbiota for symbiotic digestion. The European corn borer (ECB), Ostrinia nubilalis (Hübner), is a devastating pest that feeds the lignocellulose-rich tissues of maize plants. However, the potential role of ECB gut microbes in degrading maize cellulose remains largely unexplored. Here, we investigated the gut microbiota of ECB fed with different diets and their potential function in maize lignocellulose degradation. The diversity and composition of gut bacterial communities varied dramatically between the ECB larva fed with artificial diets (ECB-D) and maize plants (ECB-M). Draft genomes of the microbial consortia from ECB-D and ECB-M showed that the principal degraders of cellulose mainly belonged to Firmicutes or Proteobacteria and they were primarily found in the midgut. The cellulolytic microbial consortia contained genes encoding various carbohydrate-active enzymes (CAZyme). Furthermore, scanning electron microscopy revealed significant breakdown of lignocellulose in maize treated by the two microbial consortia for 9 days in vitro. Metabolomic analyses show that maize particles treated by two microbial consortia generate distinctive metabolomic profiles, with enrichment for different monosaccharides (i.e., Glucose, Rhamnofuranose, Isomaltose, and Cellobiose) and amino acids (i.e., Threonine, Histidine, and Lysine). The results indicated that the diet of the host impacted the composition and function of its gut microbiota and ECB exploited specific gut microbes to digest maize lignocellulose with distinctive products. Our study provides valuable microbiota resources for lignocellulose bioconversion.

The middle Cambrian Linyi Lagerstätte from the North China Craton: a new window on Cambrian evolutionary fauna.

The rapid appearance of major animal groups and complex marine communities during the Cambrian explosion is recorded in large part in Burgess Shale-type lagerstätten. However, the restricted temporal and spatial distribution of known lagerstätten continues to hinder the formation of a comprehensive perspective on Cambrian evolutionary faunas. Here we describe the Linyi Lagerstätte (ca. 504 mya), a new Cambrian Miaolingian lagerstätte from the Zhangxia Formation in Shandong Province, North China. The Linyi Lagerstätte contains a variety of well-preserved soft-bodied fossils, among which the non-trilobite arthropods, particularly the mollisoniids and radiodonts, are the most important groups. The new assemblage is remarkable for its excellent preservation of arthropod limbs, eyes and guts, as well as for its close similarity in taxonomic composition to Laurentian lagerstätten. The distinctive Linyi Lagerstätte holds great promise for providing additional insights into the morphological disparity, community structure and paleogeographic range of marine faunas during the middle Cambrian (Miaolingian).

Insight into Francisella tularensis-erythrocyte interactions

Francisella tularensis, the causative agent of tularemia, is a pathogenic gram-negative bacterium that enters human red blood cells (RBCs). Previous investigations suggest that invasion of RBCs helps F. tularensis survive in the gut of arthropods – a transmission vector of this bacterium leading to colonization. Since tularemia is a zoonotic infection, humans are not the primary host of F. tularensis. Therefore, we sought to investigate whether F. tularensis was capable of invading RBCs of other animals. Using the gentamicin protection assay, we discovered that while F. tularensis could enter sheep and chicken erythrocytes, bovine RBCs did not support invasion. Because of previous studies published by our lab, and due to intrinsic differences between human and bovine erythrocyte surface protein Band 3, we hypothesized that RBC membrane protein Band 3 is required for invasion. We used anti-band 3 Fab fragments to physically block the Band 3 RBC surface protein and investigated how that impacted F. tularensis invasion. Here, we provide preliminary evidence suggesting that F. tularensis uses Band 3 to enter human RBCs. This study was funded by the National Institute of Allergy and Infectious Diseases (1R15HL147135) and an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health (P20GM103434), which funds WV-INBRE program.

First record of Harpellales, Orphellales (Kickxellomycotina) and Amoebidiales (Mesomycetozoea) from Bulgaria, including a new species of Glotzia.

This paper presents the results obtained from a short survey performed in Bulgaria, southeast Europe, where the trichomycetes (sensu lato), an ecological group of arthropod gut endosymbionts, were previously completely unknown. The present study initiates the comprehension of these cryptic organisms, members of the Kickxellomycotina (Harpellales, Orphellales) and the Mesomycetozoea (Amoebidiales), in this Balkan country. Eighteen new geographic records for Bulgaria are reported, including 10 species of Harpellales, three species of Orphellales and five species of Amoebidiales. Within the Harpellales, the species Glotzia balkanensissp. nov. is described. This new species is most related to the rare species G. centroptili Gauthier ex Manier & Lichtw. and G. stenospora White & Lichtw., but is differentiated by spore and thallial characteristics. Photographs are provided and biogeographic implications of these records are discussed.

New species and geographical distribution of trichomycetes (gut fungi) in a novel host, caddisfly (Trichoptera)

Trichomycetes are fungi and protists living in the guts of arthropods. Herbivorous insects in freshwater habitats commonly house these microbes, and there are over 300 species described. One very common group of aquatic insects occurring in most freshwater systems is caddisflies, belonging to the Order Trichoptera. Although these are potentially good hosts for trichomycetes, there has been little published on trichomycetes in this host and none have been described from caddisflies. We collected and dissected caddisfly larvae from multiple sites in Canada from 2001 to 2018 and describe five new species of trichomycetes from trichopteran larvae. Data are presented on the prevalence and seasonal variation of trichomycetes in caddisfly larvae at one site in Nova Scotia where multi-year collections were made.

Aquatic Fungi: A Disregarded Trophic Level in Ecological Risk Assessment of Organic Fungicides

Freshwater fungi are a diverse group of organisms and fulfil important functions in the food web dynamics of surface water ecosystems. Ascomycetic and basidiomycetic hyphomycetes play key roles in leaf litter breakdown in rivers and creeks, while parasitic chytrids are an important food source for small invertebrates in lakes. Field studies indicate that fungal communities are affected by fungicides at environmentally relevant concentrations. However, despite their ecological importance, freshwater fungi are currently not specifically addressed in the EU regulatory frameworks with respect to the protection of surface waters. Specifically, the prospective risk assessment of fungicides does not evaluate adverse effects on non-target aquatic fungi. This paper aims to describe important functions of freshwater fungi, provides an overview of adverse effect levels of fungicides on this organism group, and proposes to integrate the fungal community of freshwater ecosystems as an additional trophic level in the current fungicide risk assessment frameworks. Results of a literature review on the effects of fungicides on aquatic fungi revealed that information on the toxicity of fungicides to non-target aquatic fungi is limited. This is, in part, due to the lack of standardised bioassays using aquatic fungi as test species. Although there is an encouraging number of bioassays focusing on the degradation of dead organic material by hyphomycetes, studies on fungicide effects on other important ecological functions, like the control of algal blooms in lentic surface waters by parasitic chytrid fungi, or on mutualistic fungi living in the guts of aquatic arthropods are largely missing. Thus, the further development and standardisation of different fungi bioassays is recommended.

A new connection between wood saprobism and beetle endosymbiosis: the rarely reported saprobic discomycete Tromeropsis is congeneric with the symbiotic yeast Symbiotaphrina (Symbiotaphrinales, Xylonomycetes) and two asexual morphs misplaced in Hyphozyma

The only hitherto known species of the monotypic genus Tromeropsis, the rarely reported T. microtheca, is redescribed from recent collections on mostly little decayed, grayed, xeric, sun-exposed wood of decorticated trunks and branches of different gymnosperms, exceptionally angiosperms, from different humid regions of central Europe and a dry area in Macaronesia. Two further, very similar species are here newly described from decayed xeric wood of different angiosperm trees and shrubs from dry to arid regions of North America, Australia, and Macaronesia. Characteristics of the genus are: black sessile apothecia, dark olivaceous exudate, fissitunicate multi-spored asci, minute, cylindrical to ellipsoid, hyaline ascospores, a yeast-like asexual morph, and, in some species, a synanamorph with allantoid conidia formed on integrated conidiogenous cells reminiscent of the genus Lecythophora but with holoblastic conidiogenesis. A lectotype is designated for the type species of the likewise sexually typified, monotypic, illegitimate genus Microspora, M. dura, which was re-examined from the original material and found to be a later synonym of T. microtheca. Nuclear rDNA data were obtained from two collections of T. microtheca and four collections of the two new species. DNA sequences of T. microtheca match those from asexual morph isolates or environmental samples in public databases. Different misapplied names attributed to sequences from asexual morph isolates gained from coniferous wood from northern Europe, North America, and eastern Asia were re-identified as T. microtheca. A very close relationship between Tromeropsis and the type species of the asexually typified genus Symbiotaphrina, S. buchneri, was observed. Independent molecular phylogenetic analyses of three rDNA regions (partial SSU, ITS, partial LSU) each place Tromeropsis and the type species of Symbiotaphrina in a single supported clade without showing clear limits between the two genera. Based on multigene analysis, Symbiotaphrina was recently placed together with the small order Xylonales in the new class Xylonomycetes. We here validate the order Symbiotaphrinales previously published as nom. nud. and describe the family Symbiotaphrinaceae. Since the name Symbiotaphrina was validated one year before Tromeropsis was published, and is more widely used, it is adopted here and T. microtheca combined into Symbiotaphrina. rDNA data further suggest that two members of the asexually typified genus Hyphozyma, H. lignicola and H. sanguinea, belong to Symbiotaphrina, whereas the type species of Hyphozyma, H. variabilis, clusters in the Thelebolales (Leotiomycetes), where it is a later synonym of Cleistothelebolus nipigonensis, the type species of Cleistothelebolus. The new combinations S. lignicola and S. sanguinea are proposed, to which the newly described species S. desertorum and S. larreae are added. Based on their close relationship and a similar yeast-like asexual morph in pure culture, we hypothesize that the life cycles of all these wood-inhabiting taxa include a symbiotic phase in the gut of arthropods and, conversely, we suppose that the life cycles of S. buchneri and S. kochii include unknown sexual morphs growing on plant substrate.

Specific and sensitive primers for the detection of predated olive fruit flies, Bactrocera oleae (Diptera: Tephritidae)

Bactrocera oleae, the olive fruit fly, is a major pest of olive (Olea europaea L.) trees worldwide. Its presence can cause important losses, with consequences for the economies of countries that produce and export table olives and olive oil. Efforts to control olive fruit fly populations have, however, been insufficient. Now more than ever, environmentally friendly alternatives need to be considered in potential control programs. Generalist predators could provide a way of managing this pest naturally. However, the identification of candidate predator species is essential if such a management system is to be introduced. The present paper describes a set of species-specific primers for detecting the presence of B. oleae DNA in the gut of predatory arthropods. All primers were tested for checking cross-reactive amplification of other fruit fly DNA and evaluated in heterospecific mixes of nucleic acids. All were found to be very sensitive for B. oleae. Subsequent feeding trials were conducted using one of the most abundant species of ground dwelling carabids in olive groves in south-eastern Madrid, Spain. These trials allowed determining that 253F-334R and 334F-253R primer pairs had the highest detection efficiency with an ID50 of around 78 h. These primers therefore provide a very useful tool for screening the gut contents of potential predators of B. oleae, and can thus reveal candidate species for the pest's biological control

Peritrophin-like protein from Litopenaeus vannamei (LvPT) involved in white spot syndrome virus (WSSV) infection in digestive tract challenged with reverse gavage

The peritrophic membrane plays an important role in the defense system of the arthropod gut. The digestive tract is considered one of the major tissues targeted by white spot syndrome virus (WSSV) in shrimp. In this study, the nucleotide sequence encoding peritrophin-like protein of Litopenaeus vannamei (LvPT) was amplified from a yeast two-hybrid library of L. vannamei. The epitope peptide of LvPT was predicted with the GenScript OptimumAntigen™ design tool. An anti-LvPT polyclonal antibody was produced and shown to specifically bind a band at ~27 kDa, identified as LvPT. The LvPT protein was expressed and its concentration determined. LvPT dsRNA (4 μg per shrimp) was used to inhibit LvPT expression in shrimp, and a WSSV challenge experiment was then performed with reverse gavage. The pleopods, stomachs, and guts were collected from the shrimp at 0, 24, 48, and 72 h post-infection (hpi). Viral load quantification showed that the levels of WSSV were significantly lower in the pleopods, stomachs, and guts of shrimp after LvPT dsRNA interference than in those of the controls at 48 and 72 hpi. Our results imply that LvPT plays an important role during WSSV infection of the digestive tract.

A PCR-based diagnostic assay for detecting DNA of the olive fruit fly, Bactrocera oleae, in the gut of soil-living arthropods.

Bactrocera oleae (Rossi) (Diptera: Tephritidae) is considered the most devastating pest of the olive tree worldwide. In an effort to develop management and biological control strategies against this pest, new molecular tools are urgently needed. In this study, we present the design of B. oleae-specific primers based on mitochondrial DNA sequences of cytochrome oxidase subunit I (COI) gene. Two pairs of B. oleae-specific primers were successfully designed and named as SBo1-F/SBo1-R and SBo2-F/SBo1-R, being able to amplify 108 and 214 bp COI fragments, respectively. The specificity of designed primers was tested by amplifying DNA from phylogenetically related (i.e. Diptera order) and other non-pest insects living in olive groves from the Mediterranean region. When using these primers on a PCR-based diagnostic assay, B. oleae DNA was detected in the gut content of a soil-living insect, Pterostichus globosus (Fabricius) (Coleoptera: Carabidae). The detection of B. oleae DNA in the guts of arthropods was further optimized by adding bovine serum albumin enhancer to the PCR reaction, in order to get a fast, reproducible and sensitive tool for detecting B. oleae remains in the guts of soil-living arthropods. This molecular tool could be useful for understanding pest-predator relationships and establishing future biological control strategies for this pest.

Orphella intropus (Kickxellomycotina), a new insect endosymbiont with an unusual perforating holdfast system and other trichomycetes from Italy

New data about trichomycetes (arthropod gut endosymbionts) from northern Italy (Piedmont [Piemonte] region) are reported, including the description of two new species of Harpellales: Glotzia distorta from mayfly nymphs and Orphella intropus from stonefly nymphs. The latter species is characterized by a cellular bulbous holdfast that perforates the gut lining of its host. Sixteen additional geographical records for Italy are provided, including from the Harpellales, Harpellomyces eccentricus, Graminella bulbosa, Orphella catalaunica, Pennella grassei, Smittium perforatum, Sm. elongatum, Stachylina nana, St. jujuyensis, St penetralis and Tectimyces leptophlebiidarum, and from the Amoebidiales, Paramoebidium chattoni, P. corpulentum, P. curvum, P. ecdyonuridae, P. hamatum and P. inflexum. We provide an emendation of Pennella grassei, a rare, incompletely described and poorly illustrated species that was recovered from Italy nearly 50 y after its last citation from France.

Acetic Acid Bacteria Genomes Reveal Functional Traits for Adaptation to Life in Insect Guts

Acetic acid bacteria (AAB) live in sugar rich environments, including food matrices, plant tissues, and the gut of sugar-feeding insects. By comparing the newly sequenced genomes of Asaia platycodi and Saccharibacter sp., symbionts of Anopheles stephensi and Apis mellifera, respectively, with those of 14 other AAB, we provide a genomic view of the evolutionary pattern of this bacterial group and clues on traits that explain the success of AAB as insect symbionts. A specific pre-adaptive trait, cytochrome bo3 ubiquinol oxidase, appears ancestral in AAB and shows a phylogeny that is congruent with that of the genomes. The functional properties of this terminal oxidase might have allowed AAB to adapt to the diverse oxygen levels of arthropod guts.