Search

Abstract Leaf-cutting ants of the genera Atta and Acromyrmex (Hymenoptera: Formicidae) are the most important pests in forest and agricultural plantations and livestock. Toxic baits are the main method to manage these insects. The objective was to determine whether the behavior of allogrooming, touch, and self-grooming among Atta sexdens rubropilosa Forel, 1908 (Hymenoptera: Formicidae) workers disperse the fungicide quinone inside inhibitor and whether this product is toxic to them. This fungicide was applied, topically, in groups of workers and the social interactions between them and their mortality with and without the fungicide were evaluated. The interactions and the quinone inside inhibitor fungicide contamination increased with the number of leaf-cutting ant workers per group. Excessive touches, with subsequent allogrooming, and self-grooming among the ant workers dispersed the quinone inside the inhibitor fungicide causing 100% mortality and indicating its toxicity to this insect. The hypothesis that social interactions contaminated ant colony mates and the toxicity of the fungicide quinone inside inhibitor to workers of the leaf-cutting ant A. sexdens rubropilosa was proven.

Considering the toxic action of P. aquilinum bracken for vertebrates, its insecticidal activity and potential use in agricultural pest control was verified, testing its effects on S. frugiperda. Crude ethanolic extract was made with the aerial part of P. aquilinum, being incorporated into artificial diet (control or extract-free, 1% and 2%) with 50 replicates (caterpillars). There was significant mortality (34% and 86%) at concentrations of 1% and 2%, respectively, compared to control (10%). In treatments at 1% (20.03 days) and 2% (20.42 days), larval stage prolongation (13.51 days) was observed in the control treatment. On the 3rd, 6th, 9th and 12th days after treatment, there was weight reduction, and the 2% treatment resulted in larvae 73% lighter than the control did at day 12. The pupae of the 2% treatment were lighter than the others. In conclusion, 1% and 2% of P. aquilinum ethanolic extract extended the larval cycle, decreased the larvae’s and pupae’s weight, and, at a concentration of 2%, caused effective mortality of S. frugiperda.

ABSTRACT Chemical control using toxic baits containing the active ingredient sulfluramid at 0.3% (w/w) is the main method for controlling leaf-cutting ants of the genera Atta and Acromyrmex. However, since 2009, when sulfluramid was included in Annex B of the Stockholm Convention on Persistent Organic Pollutants, there has been an intense search for new methods that are efficient in controlling these insects. Among said new methods, biological control using pathogenic fungi has shown promising results in laboratory conditions. The objective of this study, given the context presented, was to assess the potential of the fungi Beauveria bassiana and Trichoderma harzianum in controlling Atta sexdens. Colonies of A. sexdens were exposed to the fungi by means of formulated baits provided in a foraging chamber, or of suspensions sprayed on the fungus garden, and had their behavioral changes recorded for 21 days. For both formulations, concentrations of 10 and 20% (w/w) of the fungi being studied were used. The results allowed concluding that baits containing 10 and 20% (w/w) of the fungi B. bassiana and T. harzianum were not efficient in controlling colonies of A sexdens. On the other hand, spraying suspensions of 20% (w/w) of B. bassiana and 10% and 20% (w/w) of T. harzianum was efficient and resulted in 100% mortality of the colonies 11, 9 and 7 days after application, respectively. These findings indicate that the fungi B. bassiana and T. harzianum are promising as agents for the control of A. sexdens colonies, when sprayed on the fungus garden, although there are still some challenges as to their use related to the development of technologies for the application of the pathogen.

Leaf-cutting ants are widely distributed in Brazil, particularly species of the genus Atta. We therefore described the occurrence of leaf-cutting and grass-cutting ant species of the genus Atta. Five routes comprising some of the main highways in the South, Southeast, Center-West, and North of Brazil were sampled, in addition to ants received from other 82 municipalities, composing 300 municipalities sampled. This is the first comprehensive study of Atta in Brazil. The following species were found: A. laevigata, A. capiguara, A. sexdens rubropilosa, A. sexdens piriventris, A. sexdens sexdens and A. cephalotes. Atta laevigata and A. capiguara were the species present in the largest number of the Brazilian municipalities sampled. Atta laevigata assumed the position of species of greatest occurrence in the Brazilian territory. Atta sexdens piriventris was only recorded in the southern region of Brazil. Atta bisphaerica presented lower expansion than A. capiguara. Atta cephalotes and Atta opaciceps are species of very restricted occurrence. Southeastern region was characterized by the occurrence of A. capiguara and A. laevigata. Atta laevigata exhibited a generalized pattern of occurrence in the Center-West and North. Our study contributes to a better understanding of the so far unknown occurrence of leaf-cutting and grass-cutting ants within Brazil.

During foraging, worker ants are known for making use of many information sources to guide themselves in external environments, especially individual (memory) and social (trail pheromone) information. Both kinds of information act in a synergic way, keeping the foraging process efficient and organized. However, when social and individual information is conflicting face a trail bifurcation, it is necessary to establish a hierarchical order so prioritizing one of them. This study aims to verify which information (social or individual) is prioritized by Acromyrmex subterraneus workers when facing a bifurcation in a Y-trail system. Only one branch of the Y-trail leads to food resource and it had a section covered by filter paper where trail pheromone was deposited by workers. Pheromone deposition was here estimated by worker flow. After an individually marked forager (target-worker) made 1, 3 or 5 trips to the food resource, the filter paper was transferred to the branch which did not lead to the food. The time spent by target workers on branch selection and their right choice (branch with food) frequency were registered. Regardless of the target worker's previous trips to the resource, right choice frequency stood over 70%. In addition, the number of previous trips did not influence the time spent on decision making. However, the higher the flow of workers, the longer the time spent on decision making. By simulating a situation with conflicting information, it was possible to verify that a hierarchical order is established by A. subterraneus, which prioritized individual information (memory). Keywords: Leafcutter ants, Trail pheromone, Memory, Synergism

Atta bisphaerica is a species of grass-cutting ants commonly found in the Cerrado biome. The Brazilian Cerrado (savanna) biome covers 2 million km representing 23% of the area of the country. It is an ancient biome with rich biodiversity, estimated at 160,000 species of plants, fungi and animals. However, little is known about their nest architecture development. This study investigated the architecture of fourteen A. bisphaerica nests from Botucatu, São Paulo, Brazil. Molds were made of the nests by filling them with cement to allow better visualization of internal structures such as chambers and tunnels. After excavation, the depth and dimensions (length, width, and height) of the chambers were measured. As expected, there was a lateral development in the nests and increase in the number of chambers over time. Results showed that in nests with an estimated age of 14 months, the average depth was 1.6 ± 0.4 m; for those with 18 months it was 2.2 ± 0.7 m and at 28 months it was 2.5 ± 0.7 m. The number of chambers varied from 4 to 7 in 28-month nests, 2 to 4 in 18-month nests, and from 2 to 3 in 14-month nests. With respect to the dimensions of the internal tunnels, there were variations in their average width, increasing with time. The fungus chambers were located beneath the largest mound of loose soil. This study contributes to a better understanding of the so far unknown nest architecture development of A. bisphaerica grass-cutting ants. Keywords: Grass-cutting ants, Nest, Social insects

ABSTRACT Leaf-cutting ants live symbiotically with a fungus that they cultivate on the plant leaves that they cut. The innumerous studies on the plant selection mechanism used by leaf-cutting ants show the researchers’ interest in this issue. Many classical studies propose that plants are selected according to the fungus garden nutritional needs and the absence of potentially harmful substances. This hypothesis is corroborated by behavioral experiments using cycloheximide (fungicide) with citric pulp or forage plants greatly accepted by leaf-cutting ants. According to this hypothesis, under the action of a fungicide, the fungus emits an allomone that informs worker ants that some food is inadequate to its growth. Although some authors state that the cycloheximide “fungicide” used is specific and non toxic to ants, our findings are distinct. In our study, various concentrations of cycloheximide were administered orally to leaf-cutting worker ants in a citric pulp paste diet. After the ingestion period, the ants were isolated and offered the symbiotic fungus for 21 days and the mortality rate was evaluated. As expected, the treatment with 0.01% cycloheximide showed a low mortality rate (8.86%). At 0.1%, the mortality rate was mild (27.85%), and treatment with 1% cycloheximide resulted in moderate mortality (45.57%). In contrast, the positive control with 0.1% sulfluramid showed a high mortality rate (91.14%). Therefore, we concluded that the ingestion of high concentrations of cycloheximide results in a moderate mortality rate in leaf-cutting worker ants.

Chemical control of leaf-cutting ants is widely used, but alternative control with toxic plant extracts is promising. Substances with insecticidal potential extracted from plants have numerous ecological advantages. This study evaluated the insecticidal and/or fungicidal potential of the plants Asclepias curassavica (tropical milkweed), Rosmarinus officinalis L. (Lamiaceae) (rosemary) and Equisetum spp. (horsetail) for control of the leaf-cutting ant Atta sexdens rubropilosa Forel, 1908 (Hymenoptera: Formicidae). Forty laboratory-reared colonies of Atta sexdens rubropilosa were used. The plants were collected, dried out in a circulating air oven for 48 hours, ground, and macerated in 96o ethanol until exhaustion. After filtration, the products were evaporated under reduced pressure to obtain the ethanolic extracts. Acceptance of the reagent, topical application of the extracts, and application of baits containing 4% of the plant extracts were tested. The results showed that all plant extracts tested negatively influenced the development of the fungus garden. Baits produced with Asclepias curassavica caused the highest mortality of the colonies within 7 days. In conclusion, the ethanolic extracts of Asclepias curassavica, Rosmarinus officinalis and Equisetum spp. exhibit insecticidal (contact and ingestion) and fungicidal activity in colonies of the leaf-cutting ant Atta sexdens rubropilosa.

The success of incipient colonies of leaf-cutter ants depends on multiple factors such as temperature, humidity, soil type, the queen’s fertility and vigor, frequency of harmful microfungi, among others. Based on this observation, three factors related to Atta sexdens were characterized: (1) initial nest morphology (depth and volume); (2) queen oviposition rate; and (3) prevalence of Escovopsis (parasitic fungus) and other fungi in incipient colonies. Our observation showed a variation in chamber dimensions and volume, indicating differences in colony development rates. The oviposition rate of queens from nests of the same age showed a difference among queens. About microfungi prevalence, it was found that many are potentially deleterious. In particular, the parasite Escovopsis was present in 16.7% of the colonies. Our study shows positive correlations between queen condition and nest size/fungus garden size. These correlations collectively suggest that ecological factors such as plant availability can generate substantial variation between nests and thus shape the success of incipient colonies.

Social animals are faced with an intriguing dilemma. On the one hand, interactions between individuals are essential to exchange information and to promote cohesion, while on the other hand such interactions carry with them the risk of catching and transmitting parasites. This trade-off is particularly significant for social insects because low within-colony genetic diversity makes their colonies potentially vulnerable to parasites while frequent interactions are essential to the development of the colonial odor profile necessary for nestmate recognition. Here we investigate whether social interactions between young and old leaf-cutting ant workers show evidence of this trade-off. We find that old workers engage in more selfgrooming and mandibular scraping than young workers, both in keeping with old workers having been more exposed to parasites. In contrast, we find that young workers engaged in more allogrooming than old workers, which seems likely to have a different motivation possibly the transfer of recognition cues. Furthermore, young workers tended to engage in allogrooming with other young workers, although it was the old workers that were most active and with whom allogrooming would seem likely to optimize information or chemicals transfer. This suggests that young workers may be attempting to minimize the risk of parasite transmission during their social interactions. Although limited to behavioral data, these results hint that ant workers may be sensitive to the trade-off between the transmission of recognition cues and disease, and adjust their social interactions accordingly. Keywords: Acromyrmex, Age polyethism, Behavioral interaction, Chemicals transfer

This study investigated the stimuli that trigger digging behavior in Acromyrmex subterraneus during nest building. The hypothesis was that the presence of the fungus garden and/or brood triggers the excavation of tunnels and chambers. For the experiment, the excavation rate of individually marked workers kept in plastic cylinders filled with soil was recorded. Four treatments were applied: (1) 30 medium-sized workers, 5 g fungus garden and 30 brood items (larvae and pupae); (2) 30 medium-sized workers and 5 g fungus garden; (3) 30 medium-sized workers and 30 brood items; (4) 30 medium-sized workers without fungus and brood. After 24 h, morphological parameters of nest structure (length and width of the chambers and tunnels in cm) and the volume of excavated soil were recorded. In contrast to the expected findings, no change in morphological structure, rate of excavation by workers, or volume of excavated soil was observed between treatments, except for tunnel width, which was greater, when no brood or fungus garden was present. Thus, the results do not support the hypothesis that the fungus garden and/or brood are local stimuli for nest excavation or that they mold the internal architecture of the nest. Although this hypothesis was confirmed for Acromyrmex lundii and Atta sexdens rubropilosa, the same does not apply to A. subterraneus. The digging behavior of workers is probably the result of adaptation during nest building in different habitats. Keywords: Acromyrmex subterraneus, Digging behavior, Leaf-cutting ants, Nest building, Social insect

ABSTRACT Dynamics of the restoration of physical trails in the grass-cutting ant Atta capiguara. Leaf-cutting ants of the genus Atta build long physical trails by cutting the vegetation growing on the soil surface and removing the small objects they find across their path. Little is known on the dynamics of trail construction in these ants. How much time do they need to build a trail? To answer this question we selected six trails belonging to two different nests of A. capiguara and removed on each trail a block of soil of 20 cm × 15 cm that included a portion of the physical trail. This block was then replaced by a new block of the same size that was removed in the pasture near the trail and that was uniformly covered by the same type of vegetation as that found on the block of soil that was removed. The time required to restore the trail was then evaluated by the length of the grass blades found along the former location of the trail. The results show that ants rapidly restore the portion of the physical trail that was interrupted, which suggests that they could also do the same after their trails have been recolonized by the vegetation.

In 2009, sulfluramid, the main ingredient in toxic baits for leaf-cutting ant control, was included in Annex B of the Stockholm Convention on Persistent Organic Pollutants. This resulted in interest in the use of entomopathogenic fungi such as Beauveria bassiana and Trichoderma harzianum for leaf-cutting ant control. The efficiency of these fungi in controlling these insects and the way that ants react individually or in group to the biological risks posed by these fungi is poorly understood. For this reason, we assessed the effects of B. bassiana and T. harzianum on Atta sexdens rubropilosa larvae, pupae and workers. Moreover, we investigated whether the number of contaminated individuals within a group has an influence in controlling the spread of fungi among workers. We found that the fungus T. harzianum showed high pathogenicity against A. sexdens rubropilosa larvae and pupae, leading to faster mortality and a survival rates. On the other hand, the fungus B. bassiana was responsible for causing faster worker mortality and lower survival rates. In addition, we observed that an increase in individuals contaminated with B. bassiana or T. harzianum in the group decreases its survival rate. The results support the hypothesis that entomopathogenic fungi are efficient in controlling leaf-cutting ants when contaminated workers are allocated to groups of healthy workers.

Protein content of leaf-cutting ant queens before the nuptial flight and during the post-claustral phase. This study evaluated the crude protein content of queens of Atta sexdens before the nuptial flight and after the claustral phase in laboratory and field colonies. The hypothesis was that protein is used for survival of the queen and for early colony growth during the claustral phase. Additionally, the nest morphology, live biomass and adult population of field colonies were evaluated. Crude protein was determined by digestion of the organic material with sulfuric acid at high temperatures. The mean crude protein content was 123.23 ± 11.20 mg for females before the nuptial flight and 70.44 ± 12.21 mg for laboratory-reared queens after the claustral phase. The post-claustral crude protein content of field-collected queen was 55.90 ± 9.18 mg. With respect to the loss of crude protein as a function of duration of the claustral phase, laboratory-reared queens lost 52.79 mg and field-collected queens lost 67.33 mg compared to females before the nuptial flight. A positive linear correlation was observed between the weight of field-collected queens (256.4 ± 36.3 mg) and colony biomass (13.02 ± 9.12 g), but there was no correlation between biomass and nest depth (13.11 ± 3.82 cm). As expected, the present results support the hypothesis that protein is used for survival of the queen and for early colony growth, as demonstrated by the reduction in crude protein content as a function of duration of the claustral phase. To our knowledge, this is the first study to provide data of the dynamics of protein reserves in leaf-cutting ant queens during the claustral phase.

Energetic cost of digging behavior in workers of the leaf-cutting ant Atta sexdens (Fabricius). During nest excavation, leaf-cutting ant workers undergo reduction in their body reserve, particularly carbohydrates. In order to estimate the energetic cost of digging, groups of 30 workers of the leaf-cutting ant Atta sexdens were sealed in a hermetic chamber for 24, 48 and 72 hours, with and without soil for digging, and had the CO2 concentration measured using respirometric chambers as well as volume of soil excavated (g). As expected, the worker groups that carried out soil excavation expelled more carbon dioxide than the groups that did not excavate. Therefore, a worker with body mass of 9.65 ± 1.50 mg dug in average 0.85 ± 0.27 g of soil for 24 hours, consuming ca. 0.58 ± 0.23 J. In this study, we calculate that the energetic cost of excavation per worker per day in the experimental set-up was ca. 0.58 J.

O conhecimento do conteúdo energético das operárias é essencial para mensurar a disponibilidade energética, responsável pela manutenção e execução de atividades essenciais para o crescimento da colônia de formigas. Mas pouco se conhece sobre o conteúdo energético das operárias e quanto estará disponível para realizar atividades de, por exemplo, escavar do ninho. O presente estudo determinou o teor de lipídeo e o conteúdo energético das operárias de formigas cortadeiras inativas, antes e pós atividade de escavação. Por meio da determinação do teor de lipídeos, pode-se calcular o conteúdo energético das operárias em repouso (paradas) e compará-las com as que escavaram. O teor de lipídeo e conteúdo energético das operárias foram em média 9,1±0,8% e 111,31±54,71J, respectivamente, entretanto, as séries experimentais não diferiram significativamente. Adicionalmente, a taxa do fluxo catabólico, embasado na massa corporal das operárias foi de 14,76±10,11µW. Conclui-se que o recurso energético para a escavação do ninho não é proveniente de reserva de lipídeos corporal e, dessa forma, o conteúdo energético das operárias não alterou antes e pós-atividade de escavação.

Será que a vida em grupo acarreta uma maior atividade individual, e consequentemente, um maior gasto energético? Para responder a essa questão, hipotetizou-se que exista uma maior produção de CO2, quando se aumenta o tamanho do grupo de formigas operárias e, consequentemente, um maior custo energético ao indivíduo quando está em grupo. Assim, grupos de 10, 20, 30, 40 e 50 operárias foram fechados em uma câmara hermética, durante 24 horas. Posteriormente, efetuou-se a medição das concentrações de CO2 nos recipientes respirométricos. Ao contrário do esperado, a produção de CO2, e consequentemente, o gasto energético individual, não diferiram estatisticamente quando se aumentou o tamanho do grupo de operárias. Dessa forma, refuta-se a hipótese de que o tamanho do grupo conduz a um maior custo energético individual, haja vista a maior interação entre os indivíduos. O estudo com operárias de Atta sexdens rubropilosa evidenciou que o tamanho do grupo não conduz a um maior custo energético individual, sendo a produção de CO2 e, consequentemente, o gasto energético individual similar, independente do tamanho do grupo.

Anualmente, as colônias maduras de Atta spp. produzem formigas aladas, machos e fêmeas, que abandonam a colônia na qual foram produzidas para formarem novas colônias e, dessa forma, continuarem a perpetuação da espécie. A fundação de colônias de Atta spp. sempre envolve grande esforço da rainha, única responsável pelo cultivo do fungo e pelos cuidados consigo mesma e com sua prole. Nesse período de fundação ocorre o consumo de grande parte de suas reservas. O presente estudo investigou como o ambiente influencia o desenvolvimento das colônias recém-fundadas de Atta sexdens rubropilosa. Para tanto, foram comparadas colônias de campo coletadas cinco meses após a revoada e colônias mantidas em laboratório desde o voo nupcial. Foram analisados o peso e o tamanho da população (número de operárias, de larvas, de pupas e de ovos), o tamanho das operárias, o volume e peso do jardim de fungo e o esforço para a fundação dos ninhos (teor de lipídeos e peso das rainhas). Comparadas às colônias de campo, as colônias de laboratório apresentaram maiores valores no tamanho e no peso da população, no volume e no peso do jardim de fungo. As medidas das operárias, por classe de tamanho, não variaram entre as colônias de campo e de laboratório. O esforço da fundação dos ninhos foi maior nas colônias de campo, cujas rainhas apresentaram menor porcentagem de lipídeos e menor peso em relação às colônias de laboratório. Esses resultados demonstram que o ambiente exerce forte influência no desenvolvimento das colônias durante a fase de fundação, bem como sobre o status fisiológico das rainhas, sendo, portanto, a sobrevivência, o desenvolvimento e o sucesso da colônia reflexos das condições do ambiente e do esforço da rainha.

As formigas cortadeiras, importantes pragas agrícolas, anualmente realizam a fundação de novas colônias. Para o início de novas colônias, é preciso a escavação de túneis e de câmaras no solo, sendo que, para isso, as rainhas recém fecundadas gastam muita energia, no entanto a quantificação dessa energia gasta é desconhecida. Nossa hipótese é que a reserva lipídica, armazenada no corpo da rainha, seja utilizada durante a escavação do ninho. Testamos essa hipótese comparando o aumento do esforço de escavação das rainhas, as quais foram experimentalmente estimuladas a escavar uma, duas ou três vezes consecutivas, em relação ao controle, ou seja, em relação às rainhas que não escavaram. Foram quantificados a massa corporal e o teor de lipídios das rainhas. Os resultados mostraram que, ao contrário de nossas expectativas iniciais, massa e teor de lípideos não foram afetadas pelo aumento do esforço de escavação do ninho. Conclui-se que a escavação pelas rainhas não afetou os percentuais de lipídios em seu corpo, consequentemente, o conteúdo de energia. Provavelmente, os recursos energéticos para a escavação não se originam de fontes lipídicas, mas de outros, provavelmente carboidratos.

Os ninhos adultos das formigas cortadeiras (gênero Atta e Acromyrmex) são compostos de milhares de câmaras subterrâneas, as quais abrigam o jardim de fungo, lixo e a população desses insetos. Entretanto, como as câmaras são construídas? Para responder essa questão, nós hipotetizamos que o jardim de fungo atua como um molde para a construção das câmaras. Assim, foram utilizadas 20 colônias de 6 meses de idade, divididas em quatro séries experimentais: padrão (quantidade normal de jardim de fungo); metade de jardim de fungo; dobro de jardim de fungo e sem jardim de fungo (Testemunha). As variáveis estudadas foram: parâmetros morfológicos das estruturas (túneis e câmara formada); fluxo das atividades das operárias; volume de solo escavado. Como se esperava, o jardim de fungo atua como um molde para a construção das câmaras em formigas cortadeiras. Os resultados foram: o tratamento sem jardim de fungo não apresentou nenhuma câmara, apenas túneis, em contraposição às demais séries experimentais que apresentaram no mínimo 2 câmaras, com dimensões similares; o fluxo das operárias carregando pellet de solo por minuto durante as 72 horas diferiu estatisticamente entre as séries experimentais e, finalmente, o volume do solo escavado foi resultado da taxa de escavação das operárias, diferindo estatisticamente entre as séries experimentais. Os resultados corroboram a hipótese de que o jardim de fungo atua como um molde para a construção da câmara. A ausência de uma estrutura funcional como uma câmara, quando o jardim de fungo está ausente, comprova a hipótese.Adult nests of leaf cutting ants (genus Atta and Acromyrmex) are composed of thousands of underground chambers, which harbor the fungus garden, garbage and their population. However, how the chambers are constructed? To answer this question, we hypothesized that the fungus garden acts as a template for the chambers construction. Thus, we used 20 colonies of 6 months of age, divided into four treatments: Normal (Control); Half fungal symbiont; Double symbiont fungus and No symbiotic fungus. The variables studied were: morphology, (tunnels and chamber formed); flow of activities of workers and volume of soil excavated. As expected, treatment no symbiotic fungus did not have any cameras, just tunnels, as opposed to other treatments that showed at least two chambers, with similar dimensions. The flow of workers carrying pellet of soil per minute for 72 hours, it differed between treatments. Thus, the volume of excavated soil was the result of the excavation rate of workers, differences among the treatments. The results confirm the hypothesis that the symbiotic fungus acts as a template for the construction of the chamber. The absence of a functional structure as a chamber when is absent of symbiont fungus proves the hypothesis.

Feed adaptation is crucial for the ecological success of animals, which explore specific or varied resources according to the suitability of the selected feature. We hypothesized that the mandibles of leaf-cutting ants exhibit a greater degree of specialization because of the specific food preference of some species. The objective of this study was to highlight possible morphological differences between the mandibles of leaf-cutting ants of the genus Atta related to the food preference of the species, i.e., to investigate the morphofunctionality of mandibles in conjunction with a study of the leaf anatomy of plants selected. A detailed description of the mandibles of workers of the grass-cutting ant Atta bisphaerica and of the leaf-cutting ant Atta sexdens rubropilosa was provided, and the anatomy and chemical composition of the plant material foraged by these species were investigated. For morphometric analysis, 50 individuals of each of the four size classes were dissected for removal of the right mandible and one sample of each caste was processed for scanning electron microscopy. The Atta species had a similar mandibular morphology and differences were only observed between species and among castes, demonstrating the clear specialization of workers. Together with the anatomical study of the foraged plant material, the results indicate a high abundance of lignified cells in grasses, permitting to infer that the foraging and processing of these plants can lead to differences in behavioral acts during their preparation and incorporation in the fungus garden. We accept the hypothesis of morphofunctionality in view of the mandibular morphology observed.

Queens in the genus Atta are solely responsible for fungus cultivation and for care of herself and her offspring. Only few studies have investigated their nests in the claustral foundation and it is unknown the production rate of expelled carbon dioxide and/or of oxygen consumption in the initial colonies of leaf-cutting ants. Thus, we have studied the development of 50 initial colonies of Atta sexdens, and production of expelled carbon dioxide under laboratory conditions. The number of eggs was counted one week after nest foundation on the seventh day, the larvae counted on day 28, and the pupae between days 42 and 49. The workers emerged on the 63rd and 70th day. The CO2 concentration increased steeply in the 42nd days (20.60 ± 8.36%) and 49th days (15,37 ± 13,11 %), at 42nd days, and subsequently returned to lower values, for example, 3.35±2.84% at week seven. The present study is the first to present CO2 emission data in initial nests, in their claustral foundation under laboratory conditions.

Leaf-cutting ants rely on the obligate symbiosis with a fungus, Leucoagaricus gongylophorus, on which they feed. This fungus garden is maintained preferentially with green leaves in combination with other tissues of plant origin. The nutritional composition of this symbiotic relationship is extremely beneficial for the colony, representing the main food source for larvae and an important food component of adult workers. The objective of this study was to observe the behavioral repertoire of Atta sexdens rubropilosa during the preparation and incorporation of two different substrates, leaves and bait pellets, as well as the participation of each caste in these processes this process. The first substrate offered to the colonies were Citrus sp. leaves cut into disks and the second were pellets of citrus pulp bait formulated without the active ingredient. The observations were made under a stereomicroscope and were started when the first portion of the substrate offered was carried to the fungus garden. The behaviors executed during preparation and incorporation of the leaf disks, and subsequently of the bait pellets, were recorded using three colonies, with six repetitions of 5 and 7 hours per colony, respectively. Eight behavioral acts were recorded for the three castes found, irrespective of the substrate processed by the workers. Licking was the most frequently executed behavioral act, which occurred throughout the observation period, regardless of the substrate offered. Gardeners and generalists were the castes most engaged in the execution of this behavior. Considering the large number of individuals involved in the execution of this task, it can be concluded that this behavior is important for dispersion of the active ingredient and contamination of workers inside the colony.

Forager ants search for adequate food sources in nature and, after their discovery, decide whether the source is suitable or not for the colony. However, we asked “How do workers seek out the substrate for cultivation of the symbiontic fungus on which they feed? To answer this question, we evaluated the distance traveled by individual workers in the search of food and the distance traveled to return to the nest, as well as the time and velocity necessary for these activities. The results showed that the distance traveled by the leaf cutting ant, Atta sexdens rubropilosa (Linneus, 1758), in the search of food was greater than the distance traveled to return with the substrate to the colony. On the other hand, the mean time and velocity were similar for food search and return to the colony. These results support the hypothesis of information transfer, according to which the worker needs to return to the nest at the beginning of foraging to transfer information to other workers and thus to establish the process of worker ant foraging. It can be concluded that workers travel large distances in a random manner until finding their substrate, but the return to the nest is efficient considering the shorter distance traveled.

The nuptial flight and nest digging are high intensity activities which consume body reserves. The flight and digging effort was quantified by measuring the carbohydrate and total lipids content in males and females before and after the nuptial flight, and the queen’s digging effort during the foundation. The digging effort was quantified by experimentally stimulating the queens to dig a nest – one, two or three consecutive times – compared to the queens that did not dig. The colorimetric method was used to determine the soluble carbohydrates and extraction method of immersion was used to determine the total lipids. The results showed significant loss of carbohydrates and total lipids in males and females after the flight. On average the males contained 0.027 mg of soluble carbohydrates before the nuptial flight, and 0.005 mg after the nuptial flight, and the females contained 0.129 mg of soluble carbohydrates before the nuptial flight, and 0.079 mg after the nuptial flight. For the males the percentage of lipids decreased from 5.27±1.07% to 2.60±0.63% and for females from 36.46±4.86% to 32.62% after the nuptial flight. The digging effort of the queen caused a slight reduction in total carbohydrates, it was without digging 0.054 mg, normal digging 0.055 mg, double digging 0.045 mg (decrease of 20,22 %), and triple digging 0.044 mg (decrease of 20 %) per queen. Based on our results we conclude that the carbohydrate content is the main energetic resource used for the nuptial flight and nest digging, for males and gynes of leaf-cutting ants.

Young nests of Acromyrmex subterraneus brunneus are characterized by refuse soil in the exterior of the nest, a single fungus chamber 11 to 20 cm deep in relation to soil surface and internal volume ranging from 0.3 to 1.5 liters. These nidification patterns are important characteristics for identifying and understanding the interactions between species and their habitats.Ninhos jovens de Acromyrmex subterraneus brunneus caracterizam-se pela terra solta no exterior do ninho, por ter uma única câmara de fungo com profundidade de 11 a 20 cm em relação à superfície do solo e por ter volume interno variando de 0,3 a 1,5 litros. Estes padrões de nidificação são caracteres importantes para a identificação e o conhecimento das interações da espécie com o seu habitat.

The nuptial flight allows males and females to meet and copulate and both need energy to perform this activity. Before leaving the nest, males and females are well nourished and ready to mate. However, little is known about the lipid and energy contents in females before the nuptial flight (virgins) and after it (mated females). In this work we measured lipid concentrations in relation to body weight in these individuals. Our results showed that 16.82% of the bodies of young virgin females one month before mating flight are composed of lipids, contrasting with the 32.62% lipid content in mated females who had not excavated their nest yet, and 32.88% in those who had. The energy content measured for virgin females was 2942.63 J, contrasting with 6110.01 J for queens before excavating the nest and 5677.51 J after excavation. Based on our results, we conclude that the body mass, and therefore the lipid and energy contents in the bodies of Atta sexdens rubropilosa queens double during the last month before the nuptial flight. This energy resource is fundamental to the activities required during the nuptial flight, digging the nest and the founding of the colony.O vôo nupcial permite que machos e fêmeas se encontrem e copulem. Para isso, tanto o macho quanto a fêmea precisam de energia para efetuar tal atividade. Antes de partir do ninho de origem, machos e fêmeas estão bem nutridos e prontos para o acasalamento. Mas pouco se conhece sobre o teor de lipídeos e conteúdo energético pré-vôo nupcial (fêmeas virgens) e pós-vôo nupcial (fêmeas acasaladas). Nossos resultados indicam que as fêmeas virgens apresentaram 16,82% de lipídeos em seu corpo, enquanto as rainhas (fêmeas acasaladas) que não escavaram 32,62%, e as que escavaram 32,88%. O conteúdo energético foi de 2942,63 J para as fêmeas virgens, 6110,01 J para rainhas sem escavação e 5677,51 J para aquelas que escavaram. Com base nos resultados obtidos, concluí-se que as rainhas de Atta sexdens rubropilosa dobram a sua massa corporal, e consequentemente, o teor de lipídeos e conteúdo energético, um mês antes da revoada. Esse recurso energético é fundamental para as atividades exigidas durante o vôo nupcial, escavação do ninho e a fundação da colônia.

Leaf-cutting ants are insects that use plant material to grow fungus from which they feed. These fungus-growing ants perform various behavioral activities to establish an environment conducive to the fungus. Among these behaviors are activities that can serve to detect materials harmful to the colony, such as licking, scraping, chopping, etc. However, there are substances that may not be detected as harmful to the fungus on first contact. Cycloheximide (CHX) is one such substance, described as a fungicide that inhibits the synthesis of proteins in eukaryotes, although its mechanism of action remains unclear. The present study aimed at evaluating the behavioral changes of worker ants, time carrying orange pellets, waste deposition and mortality, when subjected to seven days of CHX-incorporated pellets and another seven days of CHX-free pellets. The fungicide adversely and structurally affected the leaf-cutter ant colonies. Their behavior went through changes, such as an increase in pellet-licking frequencies and cleaning among the ants from the third day onward. Moreover, there was an increase in time carrying orange pellets, as well as in the mortality of workers during CHX incorporation.

Leaf-cutting ant workers dig underground chambers, for housing their symbiotic fungus, interconnected by a vast quantity of tunnels whose function is to permit the entrance of food (leaves), gaseous exchanges, and movement of workers, offspring, and the queen. Digging is a task executed by a group of workers, but little is known about the group effect and group-constructed functional structures. Thus, we analyzed the structures formed by worker groups (5, 10, 20, and 40 individuals) of the leaf-cutting ant, Atta sexdens rubropilosa, for 2 days of excavation. The digging arena was the same for the 4 groups, with each group corresponding to a different density. Our results verified a pattern of tunneling by the workers, but no chamber was constructed. The group effect is well known, since the 40-worker group dug significantly more than the groups of 5, 10, and 20. These groups did not differ statistically from each other. Analysis of load/worker verified that workers of the smallest group carried the greatest load. Our paper demonstrates the group effect on the digging of nests, namely, that excavation is proportional to group size, but without emergence of a functional structure such as a chamber.

We present a detailed description of the predatory behavior of the beetle Canthon virens Mannerheim, 1829, on the leafcutter ant Atta sp. We observed 51 acts of predation, which were also recorded on film and subjected to behavioral analysis. Canthon virens exhibited 28 behaviors while predating upon Atta sp. queens. Adult beetles search for queens while flying in a zigzag pattern, 15 to 20 cm above the ground. After catching a queen, the predator stands on its back and starts cutting the queen cervix. Once the prey is decapitated, the predator rolls it until an insurmountable obstacle is reached. The distance from the site of predation to the obstacle can vary widely and is unpredictable. The beetle rolling the queen also buries it in a very peculiar way: first, it digs a small hole and pulls the queen inside, while another beetle is attached to the prey. The burial process takes many hours (up to 12) and may depend on the hardness of the soil and the presence of obstacles. In general, one or two beetles are found in a chamber with the queen after it is buried. They make the brood balls, which serve as food for the offspring. This study contributes to the knowledge of the predatory behavior of Canthon virens, a predator poorly studied in Brazil and widespread in the country.

Ant assemblages are almost all related with the vegetation composition and so can provide us important information for conservation strategies, which are especially relevant to an environmentally protected area. We sampled the ant fauna in three different phytophysionomies in order to verify if the composition of ant species is different among the areas, especially because one of the areas is a Rocky Field and there is little information about the ant fauna in this habitat. A total of 8730 individuals were registered and an NMDS analysis showed that the ant assemblies are different at the three phytophysionomies (Rocky Field, Riparian Forest, and Secondary Forest). This study shows that the species that compose the ant assemblies in different phytophysionomies are a reflex of the environment, supporting the hypothesis that the vegetational composition results in different compositions in the ant assembly. Vegetal composition is determinant in the formation of the litter and consequently in the occurrence of ant species that depend on this layer of organic matter for nesting and foraging.

ABSTRACT Atta bisphaerica is a species of grass-cutting ants commonly found in the Cerrado biome. The Brazilian Cerrado (savanna) biome covers 2 million km representing 23% of the area of the country. It is an ancient biome with rich biodiversity, estimated at 160,000 species of plants, fungi and animals. However, little is known about their nest architecture development. This study investigated the architecture of fourteen A. bisphaerica nests from Botucatu, São Paulo, Brazil. Molds were made of the nests by filling them with cement to allow better visualization of internal structures such as chambers and tunnels. After excavation, the depth and dimensions (length, width, and height) of the chambers were measured. As expected, there was a lateral development in the nests and increase in the number of chambers over time. Results showed that in nests with an estimated age of 14 months, the average depth was 1.6 ± 0.4 m; for those with 18 months it was 2.2 ± 0.7 m and at 28 months it was 2.5 ± 0.7 m. The number of chambers varied from 4 to 7 in 28-month nests, 2 to 4 in 18-month nests, and from 2 to 3 in 14-month nests. With respect to the dimensions of the internal tunnels, there were variations in their average width, increasing with time. The fungus chambers were located beneath the largest mound of loose soil. This study contributes to a better understanding of the so far unknown nest architecture development of A. bisphaerica grass-cutting ants.

Os ninhos adultos das formigas cortadeiras (gênero Atta e Acromyrmex) são compostos de milhares de câmaras subterrâneas, as quais abrigam o jardim de fungo, lixo e a população desses insetos. Entretanto, como as câmaras são construídas? Para responder essa questão, nós hipotetizamos que o jardim de fungo atua como um molde para a construção das câmaras. Assim, foram utilizadas 20 colônias de 6 meses de idade, divididas em quatro séries experimentais: padrão (quantidade normal de jardim de fungo); metade de jardim de fungo; dobro de jardim de fungo e sem jardim de fungo (Testemunha). As variáveis estudadas foram: parâmetros morfológicos das estruturas (túneis e câmara formada); fluxo das atividades das operárias; volume de solo escavado. Como se esperava, o jardim de fungo atua como um molde para a construção das câmaras em formigas cortadeiras. Os resultados foram: o tratamento sem jardim de fungo não apresentou nenhuma câmara, apenas túneis, em contraposição às demais séries experimentais que apresentaram no mínimo 2 câmaras, com dimensões similares; o fluxo das operárias carregando pellet de solo por minuto durante as 72 horas diferiu estatisticamente entre as séries experimentais e, finalmente, o volume do solo escavado foi resultado da taxa de escavação das operárias, diferindo estatisticamente entre as séries experimentais. Os resultados corroboram a hipótese de que o jardim de fungo atua como um molde para a construção da câmara. A ausência de uma estrutura funcional como uma câmara, quando o jardim de fungo está ausente, comprova a hipótese.

ABSTRACT This study investigated the stimuli that trigger digging behavior in Acromyrmex subterraneus during nest building. The hypothesis was that the presence of the fungus garden and/or brood triggers the excavation of tunnels and chambers. For the experiment, the excavation rate of individually marked workers kept in plastic cylinders filled with soil was recorded. Four treatments were applied: (1) 30 medium-sized workers, 5 g fungus garden and 30 brood items (larvae and pupae); (2) 30 medium-sized workers and 5 g fungus garden; (3) 30 medium-sized workers and 30 brood items; (4) 30 medium-sized workers without fungus and brood. After 24 h, morphological parameters of nest structure (length and width of the chambers and tunnels in cm) and the volume of excavated soil were recorded. In contrast to the expected findings, no change in morphological structure, rate of excavation by workers, or volume of excavated soil was observed between treatments, except for tunnel width, which was greater, when no brood or fungus garden was present. Thus, the results do not support the hypothesis that the fungus garden and/or brood are local stimuli for nest excavation or that they mold the internal architecture of the nest. Although this hypothesis was confirmed for Acromyrmex lundii and Atta sexdens rubropilosa, the same does not apply to A. subterraneus. The digging behavior of workers is probably the result of adaptation during nest building in different habitats.

ABSTRACT Social animals are faced with an intriguing dilemma. On the one hand, interactions between individuals are essential to exchange information and to promote cohesion, while on the other hand such interactions carry with them the risk of catching and transmitting parasites. This trade-off is particularly significant for social insects because low within-colony genetic diversity makes their colonies potentially vulnerable to parasites while frequent interactions are essential to the development of the colonial odor profile necessary for nestmate recognition. Here we investigate whether social interactions between young and old leaf-cutting ant workers show evidence of this trade-off. We find that old workers engage in more selfgrooming and mandibular scraping than young workers, both in keeping with old workers having been more exposed to parasites. In contrast, we find that young workers engaged in more allogrooming than old workers, which seems likely to have a different motivation possibly the transfer of recognition cues. Furthermore, young workers tended to engage in allogrooming with other young workers, although it was the old workers that were most active and with whom allogrooming would seem likely to optimize information or chemicals transfer. This suggests that young workers may be attempting to minimize the risk of parasite transmission during their social interactions. Although limited to behavioral data, these results hint that ant workers may be sensitive to the trade-off between the transmission of recognition cues and disease, and adjust their social interactions accordingly.

ABSTRACT During foraging, worker ants are known for making use of many information sources to guide themselves in external environments, especially individual (memory) and social (trail pheromone) information. Both kinds of information act in a synergic way, keeping the foraging process efficient and organized. However, when social and individual information is conflicting face a trail bifurcation, it is necessary to establish a hierarchical order so prioritizing one of them. This study aims to verify which information (social or individual) is prioritized by Acromyrmex subterraneus workers when facing a bifurcation in a Y-trail system. Only one branch of the Y-trail leads to food resource and it had a section covered by filter paper where trail pheromone was deposited by workers. Pheromone deposition was here estimated by worker flow. After an individually marked forager (target-worker) made 1, 3 or 5 trips to the food resource, the filter paper was transferred to the branch which did not lead to the food. The time spent by target workers on branch selection and their right choice (branch with food) frequency were registered. Regardless of the target worker's previous trips to the resource, right choice frequency stood over 70%. In addition, the number of previous trips did not influence the time spent on decision making. However, the higher the flow of workers, the longer the time spent on decision making. By simulating a situation with conflicting information, it was possible to verify that a hierarchical order is established by A. subterraneus, which prioritized individual information (memory).

ABSTRACT Atta capiguara grass-cutting ants are commonly found in the Cerrado biome, in open fields. Although grass-cutting ants build giant nests, little has been elucidated about this building pattern and when chambers and tunnels emerge. The present study describes the nest architecture development of A. capiguara grass-cutting ants from data on 31 cement-molded nests. A. capiguara nests grow with increases in the number of fungus chambers and emergence and increase of waste chambers and foraging tunnels. The structural growth of A. capiguara nests in the first year and a half of age (18 months) is vertical, with the building of the first chambers in the soil profile. After 18 months, the nests grow sideways with the addition of chambers and tunnels, and the first waste chambers appear. Between 18 and 54 months, the number of fungus chambers increases from 1-3 to 21-32, and the chambers are concentrated at the soil surface, although they can be found more than 3 m deep. In addition, the total volume of the waste chambers increases with the increment in the fungus chambers volume. Thus, this study contributes to understanding the nest architecture development of A. capiguara grass-cutting ants and demonstrates that the total volume of waste chambers is proportional to the total volume of fungus chambers suitable for the colony.