Accessory gland protein regulates pairing process and oviposition in the subterranean termite Reticulitermes chinensis after swarming.

Swarming and pairing behaviors are significant to population dispersal of termites. Tandem running is a key process in pairing behavior of dealates to find a mate. Succinylation can lead to significant changes in protein structure and function, which is widely involved in metabolism and behavior regulation in many organisms. However, whether succinylation modification regulates termites' tandem running is currently unknown. In this research, we performed quantitative modified proteomics of the subterranean termite Reticulitermes chinensis Snyder before and after alate swarming. The succinylation levels of accessory gland protein (ACP) were significantly altered after alate swarming. We found that ACP is enriched in male accessory gland and female oocytes of termites. The acetylation and succinylation sites of ACP affected tandem running of dealates. The transcriptome and metabolome analyses of alates injected with ACP and its mutant proteins showed that ß-alanine metabolism pathway was the major downstream pathway of ACP. Silencing the significantly differentially expressed genes in the ß-alanine metabolic pathway (acyl-CoA dehydrogenase, enoyl-CoA hydratase, 3-hydroxyisobutyrate dehydrogenase, methylmalonate-semialdehyde dehydrogenase) suppressed tandem running and altered oviposition of paired dealates. These findings demonstrate that protein translation modification is an important regulator of tandem running behavior of termites, which implies that the succinylation and acetylation modification sites of ACP could be potential targets for insecticide action. Our research offers a potential approach for developing novel dispersal inhibitors against social insect pests.

Robotic communication with ants.

We used a robotic gantry to test the hypothesis that tandem running in the ant Temnothorax albipennis can be successful in the absence of trail laying by the leader. Pheromone glands were placed on a pin attached to a gantry. This set-up substituted for the leader of a tandem run. Neither the pin nor the glands touched the substrate and thus the ant following the robot was tracking a plume of airborne pheromones. The robot led individual workers from their current nest to a potential new one. The robotic gantry was programmed to allow for human intervention along its path to permit the following ant to stop and survey its surroundings and then catch up with its mechanical leader. The gantry then automatically tracked the precise route taken by each ant from the new nest back to the old one. Ants led by the robot were significantly more successful at finding their way home than those we carried to the new nest that had no opportunity to learn landmarks. The robot was programmed to take either a straight or a sinusoidal path to the new nest. However, we found no significant difference in the abilities of ants that had been led on such direct or sinuous paths to find their way home. Here, the robot laid no trail but our findings suggest that, under such circumstances, the following ant may lay a trail to substitute for the missing one.

The adaptive value of tandem communication in ants: Insights from an agent-based model.

Social animals often share information about the location of resources, such as a food source or a new nest-site. One well-studied communication strategy in ants is tandem running, whereby a leader guides a recruit to a resource. Tandem running is considered an example of animal teaching because a leader adjusts her behaviour and invests time to help another ant to learn the location of a resource more efficiently. Tandem running also has costs, such as waiting inside the nest for a leader and a reduced walking speed. Whether and when these costs outweigh the benefits of tandem running is not well understood. We developed an agent-based simulation model to investigate the conditions that favour communication by tandem running during foraging. We predicted that the spatio-temporal distribution of food sources, colony size and the ratio of scouts and recruits affect colony foraging success. Our results suggest that tandem running is favoured when food sources are hard to find, differ in energetic value and are long lasting. These results mirror the findings of simulations of honeybee communication. Scouts locate food sources faster than tandem followers in some environments, suggesting that tandem running may fulfil the criteria of teaching only in some situations. Furthermore, tandem running was only beneficial above a critical colony size threshold. Taken together, our model suggests that there is a considerable parameter range that favours colonies that do not use communication by tandem running, which could explain why many ants with small colony sizes forage solitarily.

Nest Site Selection during Colony Relocation in Yucatan Peninsula Populations of the Ponerine Ants Neoponera villosa (Hymenoptera: Formicidae).

In the Yucatan Peninsula, the ponerine ant Neoponera villosa nests almost exclusively in tank bromeliads, Aechmea bracteata. In this study, we aimed to determine the factors influencing nest site selection during nest relocation which is regularly promoted by hurricanes in this area. Using ants with and without previous experience of Ae. bracteata, we tested their preference for refuges consisting of Ae. bracteata leaves over two other bromeliads, Ae. bromeliifolia and Ananas comosus. We further evaluated bromeliad-associated traits that could influence nest site selection (form and size). Workers with and without previous contact with Ae. bracteata significantly preferred this species over others, suggesting the existence of an innate attraction to this bromeliad. However, preference was not influenced by previous contact with Ae. bracteata. Workers easily discriminated between shelters of Ae. bracteata and A. comosus, but not those of the closely related Ae. bromeliifolia. In marked contrast, ants discriminated between similar sized Ae. bracteata and Ae. bromeliifolia plants, suggesting that chemical cues and plant structure play an important role. Size was also significant as they selected the largest plant when provided two dissimilar Ae. bracteata plants. Nest site selection by N. villosa workers seems to depend on innate preferences but familiarization with plant stimuli is not excluded.

Symmetry breaking and pivotal individuals during the reunification of ant colonies.

Maintenance of a social group requires the ability to reach consensus when faced with divisive choices. Thus, when migrating colonies of the ant Temnothorax rugatulus split among multiple sites, they can later reunify on the basis of queen location or differences in site quality. In this study, we found that colonies can reunify even without obvious cues to break the symmetry between sites. To learn how they do so, we observed both symmetric reunifications (between identical nests) and asymmetric reunifications (between nests of unequal quality) by colonies of individually marked ants. Both reunification types were accomplished by a tiny minority that carried nestmates from the 'losing' to the 'winning' site. Reunification effort was highly skewed in asymmetric splits, where the majority of the work was done by the first ant to transport, which nearly always came from the winning site. This contrasted with symmetric splits, where the initiator did not play an outsize role and was just as likely to come from the losing site. Symmetric reunifications were also characterized by high transporter attrition, which may help to prevent deadlocks. Tandem runs were abundant in both types and were typically led by transporters as they returned to the losing site to fetch another nestmate. Few tandem followers joined the transport effort, suggesting that tandem runs do not serve to recruit transporters but may have another, as yet unidentified role. Our results underscore the potentially large contribution of highly active individuals to group behaviour, even in decentralized societies such as ant colonies.

To reunite or not: A study of artificially fragmented Diacamma indicum ant colonies.

Social insects live together in groups and maintain cohesion to enhance their chances of survival and productivity. Colony cohesion is severely challenged during relocation. We examined the dynamics of colony reunification and the factors affecting nest choice of artificially fragmented colonies of the queenless ant Diacamma indicum. None of the twelve undisturbed colonies fragmented or relocated when a good nest was available in their neighbourhood. When colonies were artificially fragmented, they mostly (25/30) reunified into a single nest unlike in randomized time-ordered network models, indicating that reunification is not the result of random recruitment acts. When the reproductive individual was present in a good nest, the colonies reunified at this address. However, when she was present in a suboptimal nest, colonies relocated her to a better quality nest and reunified there, illustrating that quality of the new nest is more important. The work distribution and relocation dynamics of reunification were comparable to intact colonies relocating to a single new nest. This is made possible by enhanced exchange of information among tandem leaders in the form of increased number of tandem runs among them. We conclude that colony cohesion is very important and is maintained after incorporating the risks of relocation and preference for nest quality during decision making.

Characterization of recruitment through tandem running in an Indian queenless ant Diacamma indicum.

Tandem running is a primitive recruitment method employed by many ant genera. This study characterizes this behaviour during the recruitment of colony mates to a new nest in an Indian ant Diacamma indicum. Tandem leaders who have knowledge of the new nest lead a single follower at a time, to the destination by maintaining physical contact. In order to characterize tandem running, we captured and analysed 621 invitations, 217 paths and 226 termination events. Remarkably, not a single colony member was lost. While invitations were stereotypic in behaviour, termination was not. Analysis of speed revealed that the average transport speed was 4.2 cm s-1. Coupled adult-brood transport was slower than other transports but was more efficient than individual trips. Comparing tandem running with other popular recruitment methods in ants allows us to postulate that even though tandem running is primitive it is probably just another means to achieve the same end.

Network approach to understanding the organization of and the consequence of targeted leader removal on an end-oriented task.

Relocation is an important event in the lives of several social insects whereby all colony members have to be transferred to a new nest when conditions in the old nest become unfavorable. In the current study, network tools were used to examine the organization of this goal-oriented task in the Indian queenless ant Diacamma indicum which relocate their colonies by means of tandem running. Individual ants were used as nodes and tandem runs as directed edges to construct unweighted networks. Network parameters were characterized in control relocations (CRs) and in relocations where the node with the highest outdegree, that is, the Maximum tandem leader (Max TL) was experimentally removed. These were then compared to 1) randomized networks, 2) simulated networks in which Max TL was removed, and 3) simulated networks with removal of a random leader. Not only was there complete recovery of the task, but the manner in which it was organized when Max TL was removed was comparable to CRs. The results obtained from our empirical study were significantly different from the results predicted by simulations of leader removal. At an individual level, the Max TL had a significantly higher outdegree than expected by chance alone and in her absence the substitute Max TL did comparable work. In addition, the position of the Max TL in the pathway of information flow was conserved in control and experimentally manipulated conditions. Understanding the organization of this critical event as more than the sum of individual interactions using network parameters allows us to appreciate the dynamic response of groups to perturbations.

Tight knit under stress: colony resilience to the loss of tandem leaders during relocation in an Indian ant.

The movement of colonies from one nest to another is a frequent event in the lives of many social insects and is important for their survival and propagation. This goal-oriented task is accomplished by means of tandem running in some ant species, such as Diacamma indicum. Tandem leaders are central to this process as they know the location of the new nest and lead colony members to it. Relocations involving targeted removal of leaders were compared with unmanipulated and random member removal relocations. Behavioural observations were integrated with network analysis to examine the differences in the pattern of task organization at the level of individuals and that of the colony. All colonies completed relocation successfully and leaders who substituted the removed tandem leaders conducted the task at a similar rate having redistributed the task in a less skewed manner. In terms of network structure, this resilience was due to significantly higher density and outcloseness indicating increased interaction between substitute leaders. By contrast, leader-follower interactions and random removal networks showed no discernible changes. Similar explorations of other goal-oriented tasks in other societies will possibly unveil new facets in the interplay between individuals that enable the group to respond effectively to stress.

Response to a change in the target nest during ant relocation.

Decisions regarding spatial and temporal choices pertaining to a wide range of activities such as mating, feeding and resting are necessary for all organisms. Social species encounter another level of complexity, wherein inputs from multiple individuals have to be consolidated to yield a consensus. One platform on which decision making can be examined is the relocation of animal societies. Relocation is the process by which organisms move from their old dwelling to a new nest along with all the inhabitants. This exposes them to the elements and thus impacts their survival and reproduction. Diacamma indicum, the model system for our experiments is a ponerine ant that uses tandem running for colony relocation. In the present study an artificial manipulation was performed to cause a shift in the target nest. The flexibility of the relocation process and, more specifically, the response of tandem leaders to a changed target were studied. A majority of these leaders (N=262) not only re-evaluated and adapted to the change, but did so with negligible error (1.65%). This enabled colonies (N=10) to reunite at the target nest in every case. The only cost to this flexible decision making was paid in terms of additional time. Although considered to be a primitive method of recruitment, we reveal that tandem running allows D. indicum leaders to incorporate assessment of the available options at every step. This inherent flexibility in decision making would be a good strategy for organisms that need to function with incomplete information or inhabit environments that change frequently.

A trade-off between antipredatory behavior and pairing competition produced by male-male tandem running in three Reticulitermes species.

Due to the omnipresent risk of predation, termites have evolved many antipredatory behaviors. The two related species Reticulitermes speratus and R. chinensis have been demonstrated to use homosexual tandem running to decrease individual predation risk after shedding their wings. In this study, we tested risk of predation in the termite R. flaviceps, which is distantly related to the above two species. We determined that homosexual tandem running also led to low individual predation risk in dealates of R. flaviceps. Moreover, by combining a predation model with a competition model, we observed a typical trade-off phenomenon between antipredatory behavior and pairing competition produced by male-male tandem running in the above three Reticulitermes species. Our results indicated that male-male tandem running could effectively protect disadvantaged individuals from being caught, but disadvantaged individuals would be easily eliminated in pairing competition after male-male tandem running, suggesting that male-male tandem running can promote population evolution in termites by repeatedly removing the relatively inferior male individuals.

Key relocation leaders in an Indian queenless ant.

Division of labor is a central feature in social insects, wherein, simple individuals come together in groups to perform tasks that could be quite complex. It is generally believed that individuals who perform a specific task are themselves simple, interchangeable units. However, the variances in the performances of these individual insects need to be explored in greater detail. In this study, individual specialization in the context of colony relocation was examined in the Indian ponerine ant Diacamma indicum. One ant termed the maximum tandem leader (Max TL) was found to have a key role. Max TL performed 24% of the adult relocation in the colony and recruited more tandem leaders than other leaders thereby contributing to the organization of the relocation. The Max TL's role in the relocation process was further examined by comparing control relocations with experiments in which the Max TL was removed during the relocation process. Even though all the colonies relocated successfully, the relocation dynamics was significantly altered in the absence of the Max TL. We find that a single individual, the Max TL, takes up roles of a performer, organizer and catalyst during the colony relocation process, which challenges the norm that all workers are equal.