Abstract Ceratocampinae (Lepidoptera: Saturniidae) is the second most diverse subfamily of Saturniidae with 300 species described in 30 genera from southern Canada to northern Argentina. Species of this subfamily are widely distributed in Southeast Brazil, with many endemics to the Cerrado, and important as indicators of ecosystem quality. Specimens of Ceratocampinae were collected in the Parque Estadual do Mirador, Maranhão state, Brazil. Adeloneivaia acuta (Schaus, 1896), Adeloneivaia schubarti Barros & O. Mielke, 1970, Eacles penelope (Cramer, 1775) and Megaceresa pulchra (Bouvier, 1923) are recorded for the first time in Maranhão state and Northeast Brazil and Eacles fairchildi May & Oiticica, 1941, only for Maranhão. The record of five Ceratocampinae (Saturniidae) species increases the knowledge on the diversity and the importance of preserving those of this subfamily in the Cerrado biome of Maranhão state and the Northeast region of Brazil. Lepidoptera (Lepidoptera 3 Argentina quality Mirador Schaus, Schaus (Schaus 1896, 1896 , 1896) O Mielke 1970 Cramer, Cramer (Cramer 1775 Bouvier, Bouvier (Bouvier 1923 Oiticica 1941 (Saturniidae 189 197 177 192 194 18 19 17 1

Resumo Ceratocampinae (Lepidoptera: Saturniidae) é a segunda subfamília mais diversa de Saturniidae com 300 espécies descritas em 30 gêneros do sul do Canadá ao norte da Argentina. Espécies desta subfamília estão amplamente distribuídas no Sudeste do Brasil, com muitas endêmicas do Cerrado e importantes como indicadores da qualidade do ecossistema. Espécimes de Ceratocampinae foram coletados no Parque Estadual do Mirador, estado do Maranhão, Brasil. Adeloneivaia acuta (Schaus, 1896), Adeloneivaia schubarti Barros & O. Mielke, 1970, Eacles penelope (Cramer, 1775) e Megaceresa pulchra (Bouvier, 1923) são registradas pela primeira vez no estado do Maranhão e Nordeste do Brasil e Eacles fairchildi May & Oiticica, 1941 apenas para o Maranhão. O registro de cinco espécies de Ceratocampinae (Saturniidae) aumenta o conhecimento sobre a diversidade e a importância da preservação das espécies dessa subfamília no bioma Cerrado maranhense e na região Nordeste do Brasil. Lepidoptera (Lepidoptera 3 Argentina ecossistema Mirador Schaus, Schaus (Schaus 1896, 1896 , 1896) Mielke 1970 Cramer, Cramer (Cramer 1775 Bouvier, Bouvier (Bouvier 1923 Oiticica 194 (Saturniidae 189 197 177 192 19 18 17 1

RESUMO O controle químico com iscas tóxicas contendo o ingrediente ativo sulfluramida a 0,3% (p/p) é o principal método de controle de formigas cortadeiras dos gêneros Atta e Acromyrmex. No entanto, desde 2009, quando a sulfluramida foi incluída no Anexo B da Convenção de Estocolmo sobre Poluentes Orgânicos Persistentes, há uma intensa busca por novos métodos que sejam eficientes no controle desses insetos. Dentre os novos métodos, o controle biológico por fungos patogênicos tem mostrado resultados promissores em condições de laboratório. O objetivo deste estudo, dado o contexto apresentado, foi avaliar o potencial dos fungos Beauveria bassiana e Trichoderma harzianum no controle de Atta sexdens. Colônias de A. sexdens foram expostas aos fungos por meio de iscas formuladas fornecidas em câmara de forrageamento, ou de suspensões pulverizadas no jardim de fungos, e tiveram suas alterações comportamentais registradas durante 21 dias. Para ambas as formulações, foram utilizadas concentrações de 10 e 20% (p/p) dos fungos em estudo. Os resultados permitiram concluir que iscas contendo 10 e 20% (p/p) dos fungos B. bassiana e T. harzianum não foram eficientes no controle de colônias de A. sexdens. Por outro lado, a pulverização de suspensões de 20% (p/p) de B. bassiana e 10% e 20% (p/p) de T. harzianum foi eficiente e resultou em 100% de mortalidade das colônias aos 11, 9 e 7 dias após a aplicação, respectivamente. Esses achados indicam que os fungos B. bassiana e T. harzianum são promissores como agentes de controle de colônias de A. sexdens, quando pulverizados sobre o jardim de fungos, embora ainda existam alguns desafios quanto ao seu uso relacionados ao desenvolvimento de tecnologias para a aplicação do patógeno.

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.

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.

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.

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.

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 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 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 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.

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.

The knowledge of the workers energy content is essential to measure the energy availability for maintenance and performance of activities essences for the colony growth. But little is known about the workers energy content, and how much will be available to carry out activities, for example, the excavation of the nest. The present study determined the lipid content and energy content of the worker ants inactive before and after excavation activity. Through the lipids determination, it can be calculated the energy content of workers resting (standing) with those which excavated. The lipid content and energy content of the workers were on average 9.1±0.8% and energy content of the workers was on average 111.31±54.71J, respectively, however, the experimental series did not differ significantly. Additionally, catabolic flux rate based in workers body mass was 14.76±10.11µW. It was concluded that the energy resource for the excavation of the nest is not coming from reserve lipid body, and thus the energy content of the workers did not change before and after excavation activity.

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.

Does the social life entail greater individual activity, and consequently, higher energy expenditure? To answer this question, we hypothesized that there is higher CO2 production, when we increase the size of the group of workers, and hence a higher energy cost to the individual when they are in groups. Thus, groups of 10, 20, 30, 40 and 50 workers were sealed in a hermetic chamber for 24 hours. Subsequently, we performed the measurements of the CO2 concentration in the containers respirometric. Unlike the expected CO2 production, and consequently the individual energy expenditure did not differ when we increase the size of the group of workers. Thus, we refuted the hypothesis that the group size leads to a higher cost individual energy, since the greater interaction between individuals. In conclusion, our study with Atta sexdens rubropilosa workers determined that the size of the group does not lead to higher energy costs individual and CO2 production, and therefore energy expenditure similar individual, independent of the group size.

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.

Nest digging spends a lot of energy existing a question about how much energy is required to dig the tunnel and initial chamber. It was hypothesized that the lipid content is used during nest digging. This hypothesis was tested by comparing the consequences of increasing digging effort in queens that were experimentally stimulated to excavate a complete founding nest either once, twice or three times consecutively, compared to control queens that didn't dig. Weight and lipid content of queens were quantified. Results showed that, in contrast with the initial expectations, weight and lipid content were not affected by the increased digging effort in the experimentally-induced successive excavations. It was conclude that the excavation by the queens did not affect the percentages of lipids and consequently the energy content, in their bodies. Probably, energy resources for the excavation wasn't originated from lipid reserves, but from other energy sources, perhaps carbohydrates.

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.

Annually, mature colonies of Atta ants produce winged males and females, where leave the colony in which they were produced to form new colonies and thus continue the species perpetuation. The foundation of Atta spp. colonies always involves high queen effort, solely responsible for growing the fungus and the care with itself and its offspring. During the foundation there is consumption of body reserves. This study investigated how the environment influences the development of newly founded colonies of leaf cutting ant, Atta sexdens rubropilosa. Therefore, it was compared field colonies collected five months after the nuptial flight and colonies maintained in the laboratory since the nuptial flight. It was analyzed the weight and size of the population (number of workers, larvae, pupae and eggs), the workers size, the volume and weight of the fungus garden and queen effort for the nest foundation (lipid content and weight of queens). In comparison to the field colonies, the laboratory colonies showed higher values in the weight and size of population, the volume and weight of the fungus garden. Measures of workers by class size did not vary between colonies from field and laboratory. The queen effort for nest foundation was higher in field colonies, which showed a lower percentage of fat and lower weight compared to laboratory colonies. These results show that the environment has a strong influence on the colonies development during the foundation, as well as on the physiological status of the queens, and therefore the survival, development and success of the colony, it is a reflection of the queen efforts and of the environment conditions.