We surveyed 492 recent studies in the fields of ecology, evolution, and behavior (EEB) to evaluate potential for observer bias and the need for blind experimentation in each study. While 248 articles included experiments that could have been influenced by observer bias, only 13.3% of these articles indicated that experiments were blinded. The use of blind observation therefore was either grossly underreported in the surveyed articles, or many EEB studies were not blinded. We hope that a concerted effort of the field of EEB-including researchers, peer-reviewers, and journal editors-will help promote and institute routine, blind observation as an essential standard that should be practiced by all sciences.

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Group size in both multicellular organisms and animal societies can correlate with the degree of division of labour. For ants, the task specialization hypothesis (TSH) proposes that increased behavioural specialization enabled by larger group size corresponds to anatomical specialization of worker brains. Alternatively, the social brain hypothesis proposes that increased levels of social stimuli in larger colonies lead to enlarged brain regions in all workers, regardless of their task specialization. We tested these hypotheses in acacia ants (Pseudomyrmex spinicola), which exhibit behavioural but not morphological task specialization. In wild colonies, we marked, followed and tested ant workers involved in foraging tasks on the leaves (leaf-ants) and in defensive tasks on the host tree trunk (trunk-ants). Task specialization increased with colony size, especially in defensive tasks. The relationship between colony size and brain region volume was task-dependent, supporting the TSH. Specifically, as colony size increased, the relative size of regions within the mushroom bodies of the brain decreased in trunk-ants but increased in leaf-ants; those regions play important roles in learning and memory. Our findings suggest that workers specialized in defence may have reduced learning abilities relative to leaf-ants; these inferences remain to be tested. In societies with monomorphic workers, brain polymorphism enhanced by group size could be a mechanism by which division of labour is achieved.

Adequate waste management is vital for the success of social life, because waste accumulation increases sanitary risks in dense societies. We explored why different leaf-cutting ants (LCA) species locate their waste in internal nest chambers or external piles, including ecological context and accounting for phylogenetic relations. We propose that waste location depends on whether the environmental conditions enhance or reduce the risk of infection. We obtained the geographical range, habitat and refuse location of LCA from published literature, and experimentally determined whether pathogens on ant waste survived to the high soil temperatures typical of xeric habitats. The habitat of the LCA determined waste location after phylogenetic correction: species with external waste piles mainly occur in xeric environments, whereas those with internal waste chambers mainly inhabit more humid habitats. The ancestral reconstruction suggests that dumping waste externally is less derived than digging waste nest chambers. Empirical results showed that high soil surface temperatures reduce pathogen prevalence from LCA waste. We proposed that LCA living in environments unfavourable for pathogens (i.e. xeric habitats) avoid digging costs by dumping the refuse above ground. Conversely, in environments suitable for pathogens, LCA species prevent the spread of diseases by storing waste underground, presumably, a behaviour that contributed to the colonization of humid habitats. These results highlight the adaptation of organisms to the hygienic challenges of social living, and illustrate how sanitary behaviours can result from a combination of evolutionary history and current environmental conditions.

The design of transport paths in consuming entities that use routes to access food should be under strong selective pressures to reduce costs and increase benefits. We studied the adaptive nature of branching angles in foraging trail networks of the two most abundant tropical leaf-cutting ant species. We mathematically assessed how these angles should reflect the relative weight of the pressure for reducing either trail maintenance effort or traveling distances. Bifurcation angles of ant foraging trails strongly differed depending on the location of the nests. Ant colonies in open areas showed more acute branching angles, which best shorten travel distances but create longer new trail sections to maintain than a perpendicular branch, suggesting that trail maintenance costs are smaller compared to the benefit of reduced traveling distance. Conversely, ant colonies in forest showed less acute branching angles, indicating that maintenance costs are of larger importance relative to the benefits of shortening travel distances. The trail pattern evident in forests may be attributable to huge amounts of litterfall that .increase trail maintenance costs, and the abundant canopy cover that reduces traveling costs by mitigating direct sunlight and rain. These results suggest that branching angles represent a trade-off between reducing maintenance work and shortening travel distances, illustrating how animal constructions can adjust to diverse environmental conditions. This idea may help to understand diverse networks systems, including urban travel networks.

The fast touch-induced folding of leaves in sensitive plants may function for deterring herbivores, but it relies on energetically costly action potentials and interferes with photosynthesis. Here, we tested whether the intensity of the folding response in Mimosa pudica was modulated based on previous experiences, and whether the modulation was dependent on the probability of exposure to herbivores or pollinators. Younger leaves (under higher herbivory risk) reopened faster with repetitions but showed complete folding at all trials, which should be more effective as defense, but limits light exposure for longer; older leaves changed from complete to partial folding with trials, but maintained similar reopening times, which should decrease loses in photosynthesis but is less effective as defense. Unlike leaves away from inflorescences, leaves near inflorescences (i.e. more likely to be touched by flower visitors, a non-damaging stimulus) marginally decreased reopening times and shifted from complete to partial folding, a combination that decreases to the least the time leaves are light limited. All leaves showed an increased response when a new stimulus was presented after the repeated trials, suggesting that the decrease in response after repeated stimulation was not caused by mechanism exhaustion. This study shows habituation-like plasticity in a plant thigmonastic response that conforms to expectations of behavioral ecology theory usually applied to animals.

Acacia ants (Pseudomyrmex spinicola) usually prune all of the vegetation growing in the vicinity of their host plant (Acacia collinsii). This lack of branches in contact with the host tree makes the main stem of the acacia the only point of access for ants invading the resident colony. The present study assessed whether the intrusion site affects the probability of the intruder being detected. It also investigated whether ants that remain inactive on the main trunk base (a) are involved in defensive duties and (b) are morphologically different from other colony members (e.g. foraging ants). In the field, intruder ants were placed on the trunk and leaves of acacias inhabited by P. spinicola; colony members on the trunk base and the leaves were marked and monitored; and head length, head width, and thorax length of ants that were either foraging on the leaves or immobile on the trunk were measured. P. spinicola ants found intruder ants entering their host acacia via the trunk more rapidly than those entering by means of the leaves. Ants on the base of the trunk do not exchange tasks with foragers: between 5 and 47 % of the 1063 ants originally marked on the trunk were re-observed there, whereas only two of the 804 ants marked on the leaves were seen on the trunk. However, ants on the trunk were similar to foragers in morphology, suggesting that defensive ants may be part of a behavioral caste.

In social insects, worker specialization in location-related tasks could occur if they return to the same location over time. Location and task fidelity was tested in the acacia ants Pseudomyrmex spinicola, which nest inside the swollen spines of the tree, and all workers enter the spines at night and during rain. Workers were marked and followed at three locations: on the leaves, tree trunk, and ground near the plant. Behavioral tests were performed, testing the reactions of marked ants toward the food used to feed the larvae (Beltian Bodies, ''BB''), and to brood outside the spines. Marked ants and ants of known age were tested for responses to disturbance of the spines. Ants were more likely to occur in the location where they were originally marked. Trunk-marked ants discarded the BB when it had a foliole fragment attached to it, while leaf-marked ants carried it to the spine. Trunk-marked ants left larvae and exited from disturbed spines more frequently than other ants. Leaf-marked ants carried larvae and pupae more often than trunk-marked ants. Spine-marked ants left pupae more often than trunk-or leaf-marked ants. When considering age, older ants reacted aggressively when threatened, whereas younger ants protected the brood. However, younger ants reacted more aggressively when older ants were absent, and older ants were more aggressive in the presence of larvae. In sum, the spatial segregation of the ants coincided with behavioral differences, and different behavioral responses are related to the age of the ant.