Protocol to obtain long movement trajectories of leaders and followers in ant tandem runs.

Tandem running in ants is a sophisticated form of communication. Precise measurement of movement coordination by the tandem pair can shed light on social interactions. Here, we describe an integrative approach to obtain long movement trajectories of a specific tandem pair within a crowd of ants. We describe a maze-like arena, video recording and editing, and movement tracking. We integrate two pieces of image-based tracking software that have distinct individual assignment strategies. This protocol aids comparative studies of recruitment communication across species. For complete details on the use and execution of this protocol, please refer to Mizumoto et al. (2023)1 and Valentini et al. (2020).2.

Functional and mechanistic diversity in ant tandem communication.

Communication is fundamental to the organization of animal societies, often resulting in the convergent evolution of similar social behavior across lineages. However, this similarity may conceal underlying functional and mechanistic differences. Here we combined network and information-theoretic analysis to quantify how tandem recruitment is distinguishable between two ant genera, Temnothorax and Diacamma. We show that Temnothorax uses tandem running to recruit additional recruiters, while Diacamma uses it principally to move the passive majority of their colony, a task that Temnothorax accomplishes with a different behavior, social carrying. Accordingly, the network structure of Diacamma tandems was dissimilar to that of Temnothorax, instead resembling the social-carrying networks in Temnothorax. Furthermore, our information-theoretical analysis on movement trajectories revealed that Diacamma tandem runs lack bidirectional information transfer, the signature of route learning in Temnothorax. By quantifying the diversity of similar communication systems, this study increases the resolution of our understanding of animal societies.

Resilience of native ant community against invasion of exotic ants after anthropogenic disturbances of forest habitats.

The positive association between disturbances and biological invasions is a widely observed ecological pattern in the Anthropocene. Such patterns have been hypothesized to be driven by the superior competitive ability of invaders or by modified environments, as well as by the interaction of these factors. An experimental study that tests these hypotheses is usually less feasible, especially in protected nature areas. An alternative approach is to focus on community resilience over time after the anthropogenic disturbance of habitats. Here, we focused on ant communities within a forest to examine their responses after disturbance over time. We selected the Yanbaru region of northern Okinawa Island, which is a biodiversity hotspot in East Asia. We compared ant communities among roadside environments in forests where the road age differed from 5 to 25 years. We also monitored the ant communities before and after disturbance from forest thinning. We found that the species richness and abundance of exotic ants were higher in recently disturbed environments (roadsides of 5-15 years old roads), where the physical environment was warmer and drier. In contrast, the roadsides of 25-year-old roads indicated the potential recovery of the physical environment with cooler and moister conditions, likely owing to regrowth of roadside vegetation. At these sites, there were few exotic ants, except for those immediately adjacent to the road. The population density of the invasive species Technoymex brunneus substantially increased 1-2 years after forest thinning. There was no evidence of the exclusion of native ants by exotic ants that were recorded after disturbance. Our results suggest that local ant communities in the Yanbaru forests have some resilience to disturbance. We suggest that restoration of environmental components is a better strategy for maintaining native ant communities, rather than removing exotic ants after anthropogenic disturbance.

Individual experience influences reconstruction of division of labour under colony disturbance in a queenless ant species.

BACKGROUND: Division of labour (DOL) is ubiquitous across biological hierarchies. In eusocial insects, DOL is often characterized by age-related task allocation, but workers can flexibly change their tasks, allowing for DOL reconstruction in fluctuating environments. Behavioural change driven by individual experience is regarded as a key to understanding this task flexibility. However, experimental evidence for the influence of individual experience is remains sparse. Here we tested the effect of individual experience on task choice in the queenless ponerine ant, Diacamma cf. indicum from Japan. RESULTS: We confirmed that both nurses and foragers shifted to vacant tasks when the colony composition was biased to one or the other. We also found that nurses which are induced to forage readily revert to nursing when reintroduced into balanced colonies. In contrast, foragers which are induced to revert to nursing very rarely return to a foraging role, even 19 days post reintroduction to their original colony. CONCLUSIONS: Taken together, our results suggest that individual experience decreases the response threshold of original foragers, as they continue to be specialist nurses in a disturbed colony. However, original nurses do not appear strongly affected by having forager experience and revert to being nurses. Therefore, while individual experience does have an effect, other factors, such as reproductive ability, are clearly required to understand DOL maintenance in fluctuating environments.

The build-up of dominance hierarchies in eusocial insects.

Reproductive division of labour is a hallmark of eusocial insects. However, its stability can often be hampered by the potential for reproduction by otherwise sterile nest-mates. Dominance hierarchy has a crucial role in some species in regulating which individuals reproduce. Compared with those in vertebrates, the dominance hierarchies in eusocial insects tend to involve many more individuals, and should require additional selective forces unique to them. Here, we provide an overview of a series of studies on dominance hierarchies in eusocial insects. Although reported from diverse eusocial taxa, dominance hierarchies have been extensively studied in paper wasps and ponerine ants. Starting from molecular physiological attributes of individuals, we describe how the emergence of dominance hierarchies can be understood as a kind of self-organizing process through individual memory and local behavioural interactions. The resulting global structures can be captured by using network analyses. Lastly, we argue the adaptive significance of dominance hierarchies from the standpoint of sterile subordinates. Kin selection, underpinned by relatedness between nest-mates, is key to the subordinates' acceptance of their positions in the hierarchies. This article is part of the theme issue 'The centennial of the pecking order: current state and future prospects for the study of dominance hierarchies'.

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.

Male neotenic reproductives accelerate additional differentiation of female reproductives by lowering JH titer in termites.

Eusocial insects exhibit reproductive division of labor, in which only a fraction of colony members differentiate into reproductives. In termites, reproductives of both sexes are present in a colony and constantly engaged in reproduction. It has been suggested that the sex ratio of reproductives is maintained by social interactions. The presence of reproductives is known to inhibit the additional differentiation of same-sex reproductives, while it promotes the differentiation of opposite-sex reproductives. In this study, using the damp-wood termite Hodotermopsis sjostedti, physiological effects of male/female reproductives on the differentiation of supplementary reproductives (neotenics) were examined. The results showed that the only male-neotenic condition, i.e., the presence of male neotenics in the absence of female neotenics, accelerated the neotenic differentiation from female workers (i.e., pseudergates). Under this condition, the rise of juvenile hormone (JH) titer was repressed in females, and the application of a JH analog inhibited the female neotenic differentiation, indicating that the low JH titer leads to rapid differentiation. Thus, the only male-neotenic condition that actively promotes reproductive differentiation by manipulating physiological condition of females is suggested to be a mechanism underlying sexual asymmetry in reproductive function, which may lead the female-biased sex allocation of reproductives.

Social enforcement depending on the stage of colony growth in an ant.

Altruism is a paradox in Darwinian evolution. Policing is an important mechanism of the evolution and maintenance of altruism. A recently developed dynamic game model incorporating colony demography and inclusive fitness predicts that, in hymenopteran social insects, policing behaviour enforcing reproductive altruism in group members depends strongly on the colony growth stage, with strong policing as the colony develops and a relaxation of policing during the reproductive phase. Here, we report clear evidence supporting this prediction. In the ant Diacamma sp., reproduction by workers was suppressed by worker policing when the colony was small, whereas in large, mature colonies worker policing was relaxed and worker-produced males emerged. Conditional expression of traits can provide strong empirical evidence for natural selection theory if the expression pattern is precisely predicted by the theory, and our results illustrate the importance of intracolony population dynamics in the evolution of social systems.

Expansion of presoldier cuticle contributes to head elongation during soldier differentiation in termites.

In termites, the soldier caste possesses morphological features suitable for colony defence, despite some exceptions. Soldiers are differentiated via two moultings through a presoldier stage with dramatic morphogenesis. While a number of morphological modifications are known to occur during the presoldier moult, growth and morphogenesis seem to continue even after the moult. The present study, using the damp-wood termite Hodotermopsis sjostedti, carried out morphological and histological investigations on the developmental processes during the presoldier stage that is artificially induced by the application of a juvenile hormone analogue. Measurements of five body parameters indicated that head length significantly increased during the 14-day period after the presoldier moult, while it did not increase subsequently to the stationary moult (pseudergate moult as control). Histological observations also showed that the cuticular development played a role in the presoldier head elongation, suggesting that the soft and flexible presoldier cuticle contributed to the soldier morphogenesis in termites.

Social dominance and reproductive differentiation mediated by dopaminergic signaling in a queenless ant.

In social Hymenoptera with no morphological caste, a dominant female becomes an egg layer, whereas subordinates become sterile helpers. The physiological mechanism that links dominance rank and fecundity is an essential part of the emergence of sterile females, which reflects the primitive phase of eusociality. Recent studies suggest that brain biogenic amines are correlated with the ranks in dominance hierarchy. However, the actual causality between aminergic systems and phenotype (i.e. fecundity and aggressiveness) is largely unknown due to the pleiotropic functions of amines (e.g. age-dependent polyethism) and the scarcity of manipulation experiments. To clarify the causality among dominance ranks, amine levels and phenotypes, we examined the dynamics of the aminergic system during the ontogeny of dominance hierarchy in the queenless ant Diacamma sp., which undergoes rapid physiological differentiation based on dominance interactions. Brain dopamine levels differed between dominants and subordinates at day 7 after eclosion, although they did not differ at day 1, reflecting fecundity but not aggressiveness. Topical applications of dopamine to the subordinate workers induced oocyte growth but did not induce aggressiveness, suggesting the gonadotropic effect of dopamine. Additionally, dopamine receptor transcripts (dopr1 and dopr2) were elevated in the gaster fat body of dominant females, suggesting that the fat body is a potential target of neurohormonal dopamine. Based on this evidence, we suggest that brain dopamine levels are elevated in dominants as a result of hierarchy formation, and differences in dopamine levels cause the reproductive differentiation, probably via stimulation of the fat body.

Global network structure of dominance hierarchy of ant workers.

Dominance hierarchy among animals is widespread in various species and believed to serve to regulate resource allocation within an animal group. Unlike small groups, however, detection and quantification of linear hierarchy in large groups of animals are a difficult task. Here, we analyse aggression-based dominance hierarchies formed by worker ants in Diacamma sp. as large directed networks. We show that the observed dominance networks are perfect or approximate directed acyclic graphs, which are consistent with perfect linear hierarchy. The observed networks are also sparse and random but significantly different from networks generated through thinning of the perfect linear tournament (i.e. all individuals are linearly ranked and dominance relationship exists between every pair of individuals). These results pertain to global structure of the networks, which contrasts with the previous studies inspecting frequencies of different types of triads. In addition, the distribution of the out-degree (i.e. number of workers that the focal worker attacks), not in-degree (i.e. number of workers that attack the focal worker), of each observed network is right-skewed. Those having excessively large out-degrees are located near the top, but not the top, of the hierarchy. We also discuss evolutionary implications of the discovered properties of dominance networks.