Humoral and Cellular Defense Mechanisms in Rebel Workers of Apis mellifera

The physiological state of an insect depends on efficiently functioning immune mechanisms such as cellular and humoral defenses. However, compounds participating in these mechanisms also regulate reproductive caste formation and are responsible for reproductive division of labor as well as for labor division in sterile workers. Divergent reaction of the same genotype yielding reproductive queens and worker castes led to shaping of the physiological and behavioral plasticity of sterile or reproductive workers. Rebels that can lay eggs while maintaining tasks inside and outside the colony exhibit both queen and worker traits. So, we expected that the phagocytic index, JH3 titer, and Vg concentration would be higher in rebels than in normal workers and would increase with their age. We also assumed that the numbers of oenocytes and their sizes would be greater in rebels than in normal workers. The rebels and the normal workers were collected at the age of 1, 7, 14, and 21 days, respectively. Hemolymph and fat bodies were collected for biochemical and morphological analyses. The high levels of JH, Vg, and the phagocytic index, as well as increased numbers and sizes of oenocytes in the fat body cells demonstrate the physiological and phenotypic adaptation of rebels to the eusocial life of honeybees.

Worker-dependent gut symbiosis in an ant

The hallmark of eusocial insects, honeybees, ants, and termites, is division of labor between reproductive and non-reproductive worker castes. In addition, environmental adaption and ecological dominance are also underpinned by symbiotic associations with beneficial microorganisms. Microbial symbionts are generally considered to be maintained in an insect colony in two alternative ways: shared among all colony members or inherited only by a specific caste. Especially in ants, the reproductive caste plays a crucial role in transmission of the symbionts shared among colony members over generations. Here, we report an exceptional case, the worker-dependent microbiota in an ant, Diacamma cf. indicum from Japan. By collecting almost all the individuals from 22 colonies in the field, we revealed that microbiota of workers is characterized by a single dominant bacterium localized at the hindgut. The bacterium belonging to an unclassified member within the phylum Firmicutes, which is scarce or mostly absent in the reproductive castes. Furthermore, we show that the gut symbiont is acquired at the adult stage. Collectively, our findings strongly suggest that the specific symbiont is maintained by only workers, demonstrating a novel pattern of ant-associated bacterial symbiosis, and thus further our understanding of host-microbe interactions in the light of sociobiology.

The role of epigenetics, particularly DNA methylation, in the evolution of caste in insect societies

Eusocial insects can be defined as those that live in colonies and have distinct queens and workers. For most species, queens and workers arise from a common genome, and so caste-specific developmental trajectories must arise from epigenetic processes. In this review, we examine the epigenetic mechanisms that may be involved in the regulation of caste dimorphism. Early work on honeybees suggested that DNA methylation plays a causal role in the divergent development of queen and worker castes. This view has now been challenged by studies that did not find consistent associations between methylation and caste in honeybees and other species. Evidence for the involvement of methylation in modulating behaviour of adult workers is also inconsistent. Thus, the functional significance of DNA methylation in social insects remains equivocal. This article is part of the theme issue ‘How does epigenetics influence the course of evolution?’

Regulation of oogenesis in the queen honey bee (Apis mellifera)

In the honey bee (Apis mellifera), queen and worker castes originate from identical genetic templates but develop into different phenotypes. Queens lay up to 2,000 eggs daily whereas workers are sterile in the queen’s presence. Periodically queens stop laying; during swarming, when resources are scarce in winter and when they are confined to a cage by beekeepers. We used confocal microscopy and gene expression assays to investigate the control of oogenesis in honey bee queen ovaries. We show that queens use different combination of ‘checkpoints’ to regulate oogenesis compared to honey bee workers and other insect species. However, both queen and worker castes use the same programmed cell death pathways to terminate oocyte development at their caste-specific checkpoints. Our results also suggest that the termination of oogenesis in queens is driven by nutritional stress. Thus, queens may regulate oogenesis via the same regulatory pathways that were utilised by ancestral solitary species but have adjusted physiological checkpoints to suit their highly-derived life history.Summary statementHoney bee queens regulate oogenesis using a different combination of ‘checkpoints’ to workers, but both castes use the same molecular pathways.

Genetic toolkit for sociality predicts castes across the spectrum of social complexity in wasps

Major evolutionary transitions describe how biological complexity arises; e.g. in evolution of complex multicellular bodies, and superorganismal insect societies. Such transitions involve the evolution of division of labour, e.g. as queen and worker castes in insect societies. Castes across different evolutionary lineages are thought to be regulated by a conserved genetic toolkit. However, this hypothesis has not been tested thoroughly across the complexity spectrum of the major transition. Here we reveal, using machine learning analyses of brain transcription, evidence of a shared genetic toolkit across the spectrum of social complexity in Vespid wasps. Whilst molecular processes underpinning the simpler societies (which likely represent the origins of social living) are conserved throughout the major transition, additional processes appear to come into play in more complex societies. Such fundamental shifts in regulatory processes with complexity may typify other major evolutionary transitions, such as the evolution of multicellularity.

The diversification of termites: inferences from a complete species-level phylogeny

Termites play a major role in a variety of ecological processes in tropical and subtropical biomes worldwide, such as decomposition, soil formation and aeration, and nutrient cycling. These important ecosystem services were achieved through their highly complex societies and remarkable adaptations, including the evolution of sterile worker castes, the acquisition of endosymbionts, and the capacity for extensive environmental engineering, yet the causes and consequences of their ecological success are still poorly understood. The goals of our study were (1) to provide the first complete, species-level phylogeny of all currently recognized termite species by integrating the available genetic and taxonomic data, as well as methods of phylogenetic imputation and divergence time estimation; and (2) to explore variation in speciation rates among termite lineages. We provide the inferred relationships as a set of 1,000 pseudo-posterior trees, which can be used in future comparative analyses. We demonstrate that speciation rates have been relatively constant throughout the history of termites, with two positive shifts in speciation rates: one at their origin of Euisoptera and the other concordant with evolution of Termitidae. On the other hand, there was no obvious trend towards deceleration in speciation rates for termites as a whole, nor within the most species-rich families. The provided trees might represent a valuable resource for termite comparative studies by summarizing the available phylogenetic information, while accounting for uncertainty in the inferred topologies.

Mandibular morphology, task specialization, and bite mechanics in Pheidole ants (Hymenoptera: Formicidae)

The remarkable ecological and evolutionary success of ants was associated with the evolution of reproductive division of labor, in which sterile workers perform most colony tasks whereas reproductives become specialized in reproduction. In some lineages, the worker force became further subdivided into morphologically distinct subcastes (e.g. minor vs. major workers), allowing for the differential performance of particular roles in the colony. However, the functional and ecological significance of morphological differences between subcastes is not well understood. Here, we applied Finite Element Analysis (FEA) to explore the functional differences between major and minor ant worker mandibles. Analyses were carried out on mandibles of two Pheidole species, a dimorphic ant genus. In particular, we test whether major mandibles evolved to minimize stress when compared to minors using combinations of tooth and masticatory margin bites under strike and pressure conditions. Majors performed better in pressure conditions yet, contrary to our expectations, minors performed better in strike bite scenarios. Moreover, we demonstrate that even small morphological differences in ant mandibles might lead to substantial differences in biomechanical responses to bite loading. These results also underscore the potential of FEA to uncover biomechanical consequences of morphological differences within and between ant worker castes.

Alcohol intoxication resistance and alcohol dehydrogenase levels differ between the honeybee castes

Various animal models are used in the study of alcoholism, with the honeybee (Apis mellifera L.) among them. Here, we tested the hypothesis that foragers show higher intoxication resistance to alcohol than nurses, an issue thus far not investigated. To this end, we measured the latency to full sedation when exposed to alcohol in foragers, nurses and reverted nurses. In addition, we measured alcohol dehydrogenase (ADH) levels in these worker castes. Caste status was confirmed by comparison of the size of their hypopharyngeal glands. We detected high intoxication resistance to alcohol and presence of ADH in foragers. In nurses, we detected significantly lower intoxication resistance to alcohol and no ADH. These between-caste differences cannot be explained by the age difference between castes as in reverted nurses, characterized by similar age to foragers, we detected an intermediate intoxication resistance to alcohol and no ADH. Our results suggest possible natural exposure to alcohol in different castes of workers. As such, we further develop the honeybee as a model in alcoholism-related research and open new research avenues.

A new nasute-termite Ceylonitermes nivedita sp. nov. (Isoptera: Termitidae) from India along with first record of worker dimorphism

A new nasute termite Ceylonitermes nivedita sp. nov. (Isoptera: Termitidae) is described from the tropical rainforests of Manipur, India. Both the soldier and worker castes of the species are described. The workers are strongly dimorphic and no such dimorphism is reported hitherto among the 3 known species of Ceylonitermes. A revised dichotomous key to identify all the 4 known species of the genus is also provided. The proposed new species is a minor pest of bamboo. Key words: Ceylonitermes, India, Nasutitermitinae, key