An experimental, behavioral, and chemical analysis of food limitations in mutualistic Crematogaster ant symbionts inhabiting Macaranga host plants

Obligate mutualistic plant-ants are often constrained by their plant partner's capacity to provide resources. However, despite this limitation, some ant partners actively reject potential prey items and instead drop them from the plant rather than consuming them, leaving the ants entirely reliant on host plant-provided food, including that provided indirectly by the symbiotic scale insects that ants tend inside the plants. This dependency potentially increases the efficiency of these ants in defending their host. We hypothesize that if this ant behavior was beneficial to the symbiosis, prey rejection by ants would be observed across multiple plant host species. We also hypothesize that plant-provided food items and symbiotic scale insects from other ant plants should be rejected. We address these hypotheses in the Crematogaster ant-Macaranga plant system, in which plants provide living space and food, while ants protect plants from herbivory. We observed food acceptance and rejection behavior across five ant species and three plant host species. Ants were offered three types of food: termites as a surrogate herbivore, symbiotic scale insects, and nutritious food bodies (FB) produced by different host plant species. The unique ant species living in M. winkleri was the most likely to reject food items not provided by the plant species, followed by ants in M. glandibracteolata, while ants in M. pearsonii accepted most items offered to them. Using stable isotopes, chemical cues, and proteomic analyses, we demonstrate that this behavior was not related to differences between plant species in nutritional quality or composition of FB. Isotopic signatures revealed that certain species are primary consumers but other ant species can be secondary consumers even where surrogate herbivores are rejected, although these values varied depending on the ant developmental stage and plant species. Macaranga pearsonii and M. glandibracteolata, the two most closely related plant species, had most similar surface chemical cues of FB. However, M. glandibracteolata had strongest differences in food body nutritional content, isotopic signatures, and protein composition from either of the other two plant species studied. Taken together we believe our results point toward potential host coercion of symbiont ants by plants in the genus Macaranga Thouars (Euphorbiaceae)

Arboreal ant abundance tracks primary productivity in an Amazonian whitewater river system

Little is known about consumer productivity in the tropics despite the key feedbacks that animals impose on primary productivity. In the Amazon basin, seasonally flooded and unflooded forests exist side by side, and ants (Formicidae) dominate animal biomass. Although flooding has a direct negative effect on soil-dwelling ants, it is less clear whether flooding has indirect effects on arboreal ants via associated changes in tree communities. To test whether seasonal inundation by whitewater affects arboreal ants, we investigated ant communities in adjacent flooded and unflooded forests along a major whitewater river in central-western Amazonia. Whitewater-flooded forest exhibits higher primary productivity than unflooded forest. We thus hypothesized that forest type would affect the productivity and the foraging traits of arboreal ants and that these changes would be mediated by increases in plant-derived food for ants in flooded forest. We compared ant and plant communities between flooded- and unflooded-forest transects along the Juruá River in Amazonas, Brazil. We collected, identified, and counted terrestrial and arboreal ants, and we measured ant traits with putative relationships to foraging strategy. We also identified plant stems to characterize the abundance of ant food rewards. Flooding negatively affected the diversity and abundance of terrestrial ants but did not change the diversity of arboreal ants. Arboreal ants were more abundant and exhibited higher biomass in flooded forest than in unflooded forest. Arboreal ant traits also suggested that ants may rely more heavily on plant-derived food in flooded forest than in unflooded forest. These differences were associated with a higher abundance of plant stems predicted to contain ant food rewards in flooded forest than in unflooded forest. Our results indicate that the productivity of arboreal ants is affected by that of the underlying forest. Such effects may be mediated by the predominantly herbivorous foraging strategy of canopy ants, which would link ant populations closely to primary production and stoichiometry. Given ants’ important functional roles, these differences in ant productivity between forest types may have consequences for other arthropods and feedbacks to plants throughout the Amazon basin

Genomic changes associated with the evolutionary transition of an insect gut symbiont into a blood-borne pathogen.

The genus Bartonella comprises facultative intracellular bacteria with a unique lifestyle. After transmission by blood-sucking arthropods they colonize the erythrocytes of mammalian hosts causing acute and chronic infectious diseases. Although the pathogen-host interaction is well understood, little is known about the evolutionary origin of the infection strategy manifested by Bartonella species. Here we analyzed six genomes of Bartonella apis, a honey bee gut symbiont that to date represents the closest relative of pathogenic Bartonella species. Comparative genomics revealed that B. apis encodes a large set of vertically inherited genes for amino acid and cofactor biosynthesis and nitrogen metabolism. Most pathogenic bartonellae have lost these ancestral functions, but acquired specific virulence factors and expanded a vertically inherited gene family for harvesting cofactors from the blood. However, the deeply rooted pathogen Bartonella tamiae has retained many of the ancestral genome characteristics reflecting an evolutionary intermediate state toward a host-restricted intraerythrocytic lifestyle. Our findings suggest that the ancestor of the pathogen Bartonella was a gut symbiont of insects and that the adaptation to blood-feeding insects facilitated colonization of the mammalian bloodstream. This study highlights the importance of comparative genomics among pathogens and non-pathogenic relatives to understand disease emergence within an evolutionary-ecological framework

Strategies of a parasite of the ant–Acacia mutualism

Mutualisms can be exploited by parasites—species that obtain resources from a partner but provide no services. Though the stability of mutualisms in the presence of such parasites is under intensive investigation, we have little information on life history traits that allow a species to be a successful mutualist or rather a parasite, particularly in cases where both are closely related. We studied the exploitation of Acacia myrmecophytes by the ant, Pseudomyrmex gracilis, contrasting with the mutualistic ant Pseudomyrmex ferrugineus. P. gracilis showed no host-defending behavior and had a negative effect on plant growth. By preventing the mutualist from colonization, P. gracilis imposes opportunity costs on the host plant. P. gracilis produced smaller colonies with a higher proportion of alates than did the mutualist and thus showed an “r-like” strategy. This appears to be possible because P. gracilis relies less on host-derived food resources than does the mutualist, as shown by behavioral and stable isotope studies. We discuss how this system allows the identification of strategies that characterize parasites of mutualisms

Evolutionary Convergence and Nitrogen Metabolism in Blattabacterium strain Bge, Primary Endosymbiont of the Cockroach Blattella germanica

Bacterial endosymbionts of insects play a central role in upgrading the diet of their hosts. In certain cases, such as aphids and tsetse flies, endosymbionts complement the metabolic capacity of hosts living on nutrient-deficient diets, while the bacteria harbored by omnivorous carpenter ants are involved in nitrogen recycling. In this study, we describe the genome sequence and inferred metabolism of Blattabacterium strain Bge, the primary Flavobacteria endosymbiont of the omnivorous German cockroach Blattella germanica. Through comparative genomics with other insect endosymbionts and free-living Flavobacteria we reveal that Blattabacterium strain Bge shares the same distribution of functional gene categories only with Blochmannia strains, the primary Gamma-Proteobacteria endosymbiont of carpenter ants. This is a remarkable example of evolutionary convergence during the symbiotic process, involving very distant phylogenetic bacterial taxa within hosts feeding on similar diets. Despite this similarity, different nitrogen economy strategies have emerged in each case. Both bacterial endosymbionts code for urease but display different metabolic functions: Blochmannia strains produce ammonia from dietary urea and then use it as a source of nitrogen, whereas Blattabacterium strain Bge codes for the complete urea cycle that, in combination with urease, produces ammonia as an end product. Not only does the cockroach endosymbiont play an essential role in nutrient supply to the host, but also in the catabolic use of amino acids and nitrogen excretion, as strongly suggested by the stoichiometric analysis of the inferred metabolic network. Here, we explain the metabolic reasons underlying the enigmatic return of cockroaches to the ancestral ammonotelic state

Heritable symbionts in a world of varying temperature

Heritable microbes represent an important component of the biology, ecology and evolution of many plants, animals and fungi, acting as both parasites and partners. In this review, we examine how heritable symbiont–host interactions may alter host thermal tolerance, and how the dynamics of these interactions may more generally be altered by thermal environment. Obligate symbionts, those required by their host, are considered to represent a thermally sensitive weak point for their host, associated with accumulation of deleterious mutations. As such, these symbionts may represent an important determinant of host thermal envelope and spatial distribution. We then examine the varied relationship between thermal environment and the frequency of facultative symbionts that provide ecologically contingent benefits or act as parasites. We note that some facultative symbionts directly alter host thermotolerance. We outline how thermal environment will alter the benefits/costs of infection more widely, and additionally modulate vertical transmission efficiency. Multiple patterns are observed, with symbionts being cold sensitive in some species and heat sensitive in others, with varying and non-coincident thresholds at which phenotype and transmission are ablated. Nevertheless, it is clear that studies aiming to predict ecological and evolutionary dynamics of symbiont–host interactions need to examine the interaction across a range of thermal environments. Finally, we discuss the importance of thermal sensitivity in predicting the success/failure of symbionts to spread into novel species following natural/engineered introduction

Genome-Wide Functional Divergence after the Symbiosis of Proteobacteria with Insects Unraveled through a Novel Computational Approach

Symbiosis has been among the most important evolutionary steps to generate biological complexity. The establishment of symbiosis required an intimate metabolic link between biological systems with different complexity levels. The strict endo-cellular symbiotic bacteria of insects are beautiful examples of the metabolic coupling between organisms belonging to different kingdoms, a eukaryote and a prokaryote. The host (eukaryote) provides the endosymbiont (prokaryote) with a stable cellular environment while the endosymbiont supplements the host's diet with essential metabolites. For such communication to take place, endosymbionts' genomes have suffered dramatic modifications and reconfigurations of proteins' functions. Two of the main modifications, loss of genes redundant for endosymbiotic bacteria or the host and bacterial genome streamlining, have been extensively studied. However, no studies have accounted for possible functional shifts in the endosymbiotic proteomes. Here, we develop a simple method to screen genomes for evidence of functional divergence between two species clusters, and we apply it to identify functional shifts in the endosymbiotic proteomes. Despite the strong effects of genetic drift in the endosymbiotic systems, we unexpectedly identified genes to be under stronger selective constraints in endosymbionts of aphids and ants than in their free-living bacterial relatives. These genes are directly involved in supplementing the host's diet with essential metabolites. A test of functional divergence supports a strong relationship between the endosymbiosis and the functional shifts of proteins involved in the metabolic communication with the insect host. The correlation between functional divergence in the endosymbiotic bacterium and the ecological requirements of the host uncovers their intimate biochemical and metabolic communication and provides insights on the role of symbiosis in generating species diversity

Bacterial Gut Symbionts Contribute to Seed Digestion in an Omnivorous Beetle

Obligate bacterial symbionts alter the diets of host animals in numerous ways, but the ecological roles of facultative bacterial residents that colonize insect guts remain unclear. Carabid beetles are a common group of beneficial insects appreciated for their ability to consume insect prey and seeds, but the contributions of microbes to diet diversification in this and similar groups of facultative granivores are largely unknown.Using 16S rRNA gene clone libraries and terminal restriction fragment (tRF) length polymorphism analyses of these genes, we examined the bacterial communities within the guts of facultatively granivorous, adult Harpalus pensylvanicus (Carabidae), fed one of five dietary treatments: 1) an untreated Field population, 2) Seeds with antibiotics (seeds were from Chenopodium album), 3) Seeds without antibiotics, 4) Prey with antibiotics (prey were Acheta domesticus eggs), and 5) Prey without antibiotics. The number of seeds and prey consumed by each beetle were recorded following treatment. Harpalus pensylvanicus possessed a fairly simple gut community of approximately 3-4 bacterial operational taxonomic units (OTU) per beetle that were affiliated with the Gammaproteobacteria, Bacilli, Alphaproteobacteria, and Mollicutes. Bacterial communities of the host varied among the diet and antibiotic treatments. The field population and beetles fed seeds without antibiotics had the closest matching bacterial communities, and the communities in the beetles fed antibiotics were more closely related to each other than to those of the beetles that did not receive antibiotics. Antibiotics reduced and altered the bacterial communities found in the beetle guts. Moreover, beetles fed antibiotics ate fewer seeds, and those beetles that harbored the bacterium Enterococcus faecalis consumed more seeds on average than those lacking this symbiont.We conclude that the relationships between the bacterium E. faecalis and this factultative granivore's ability to consume seeds merit further investigation, and that facultative associations with symbiotic bacteria have important implications for the nutritional ecology of their hosts

Ecological Indicator Values of Europe (EIVE) 1.0: a powerful open-access tool for vegetation scientists

Background: Ecological indicator values (EIVs) have a long tradition in vegetation ecological research in Europe. EIVs characterise the ecological optimum of species along major environmental gradients using ordinal scales. Calculating mean indicator values per plot is an effective way of bioindication. Following first systems in Russia and Central Europe, about two dozen EIV systems have been published for various parts of Europe. Aims: As there was no EIV system available at European scale that could be used for broad-scale analyses, e.g. in the context of the European Vegetation Archive (EVA), we develop such a system for the first time for the vascular plants of Europe. Location: Europe. Methods: We compiled all national and major regional EIV systems and harmonized their plant nomenclature with a newly developed contemporary European taxonomic backbone (EuroSL 1.0). Using regression, we rescaled the individual EIV systems for the main parameters to continent-wide quasi-metric scales, ranging from 1 to 99. The data from each individual system were then translated into a probability curve approximated with a normal distribution, weighed with the logarithm of the area represented and summed up across the systems. From the European density curve we extracted then a mean and a variance, which characterise the distribution of this species along this particular ecological gradient. Results and conclusions: Our consensus approach of integrating the expert knowledge of all existing EIV systems allowed deriving the first consistent description of the ecological behaviour for a significant part of the European vascular flora. The resulting Ecological Indicator Values of Europe (EIVE) 1.0 will be published open access to allow bioindication beyond country borders. Future releases of EIVE might contain more parameters, non-vascular plants and regionalisation or could be re-adjusted and extended to hitherto non-covered species through co-occurrence data from EVA