Perceptual differences in trail-following leaf-cutting ants relate to body size

Leaf-cutting ants of the genus Atta have highly size-polymorphic workers, and size is related to division of labor. We studied trail-following behavior of different-sized workers in a laboratory colony of Atta vollenweideri. For small and large workers, we measured responsiveness and preference to artificial conspecific and heterospecific pheromone trails made from poison gland extracts of A. vollenweideri and A. sexdens. Responsiveness was measured as the probability of trail-following, and preference was measured by testing the discrimination between one conspecific and one heterospecific trail. Minute amounts of the releaser component methyl-4-methylpyrrole-2-carboxylate (0.4pg/1m), present in both, conspecific and heterospecific trails, suffice to elicit trail-following behavior. Workers followed heterospecific trails, and these trails (after normalizing their concentration) were as effective as conspecific trails. Small workers were less likely to follow a trail of a given concentration than large workers. In the discrimination test, small workers preferred the conspecific trail over the heterospecific trail, whereas large workers showed no significant preference. It is suggested that large workers primarily respond to the releaser component present in both trails, whereas small workers focus more on the conspecific traits provided by the blend of components contained in the trail pheromone.     

Chemical and behavioural studies of the trail-following pheromone in the leaf-cutting ant Atta opaciceps,Borgmeier (Hymenoptera: Formicidae)

To understand the significance of the trail pheromone used in chemical communication of the leaf-cutting ants Atta opaciceps we investigated, under laboratory conditions, the trail-following behaviour of different castes. We observed a clear behavioural discrimination of conspecific venom gland extract of foraging ants from those of other species. Additionally, we determined the pheromone composition of A. opaciceps venom gland secretion using a two-dimensional gas chromatography coupled with mass spectrometry. Chemical analyses revealed the presence of three nitrogen-containing compounds, identified as 2,5-dimethylpyrazine, 3-ethyl-2,5-dimethylpyrazine and methyl 4-methylpyrrole-2-carboxylate (M4MPC). Four different bioassays performed with workers from different castes of A. opaciceps suggested that the trail pheromone elicits the trail-following behaviour in conspecifics of all castes, but the foragers respond more strongly to their own pheromone than to that of other castes (gardeners, generalists and soldiers). In addition, A. opaciceps foragers follow the trails made with the venom gland extracts of the unrelated Acromyrmex subterraneus subterraneus foragers as well as they follow the trails made with their own venom gland extract. M4MPC was identified to be the most abundant and the most behaviourally active component of the venom gland extract of A. opaciceps foragers.     

Polyethism and nestmate recognition in the alarm reaction of Atta leaf-cutting ants

Abstract.  Ethological studies are conducted under laboratory conditions using workers of Atta sexdens sexdens and Atta opaciceps collected from field colonies to investigate the mechanisms involved in the alarm response and intra-specific recognition in leaf-cutting ants. Hexane extracts from the heads of gardeners and generalist workers elicit higher levels of alarm response in foragers from the same colony than do mandibular gland extracts from foragers and soldiers, indicating that gardeners, generalists and foragers are primarily responsible for the production of alarm pheromone. Foragers subjected to extracts from non-nestmates exhibit significantly greater alarm responses than are induced by similar extracts derived from nestmate workers, suggesting that the alarm pheromone may have a role in nestmate recognition.     

Survey of Neodohrniphora spp. (Diptera: Phoridae) at colonies of Atta sexdens rubropilosa (FOREL) and specificity of attack behaviour in relation to their hosts

Atta sexdens rubropilosa is a leaf-cutting ant that is a significant agricultural and forestry pest in the Neotropical region. This ant is parasitized by flies from the genera Neodohrniphora spp., Apocephalus spp. and Myrmosicarius spp. This study was carried out to determine which species of Neodohrniphora spp. are found near foraging trails of Atta sexdens rubropilosa and to evaluate the specificity of attack behaviour of these parasitoids. From May 2002 to April 2004, we sampled Neodohrniphora spp. hovering over foraging trails of Atta sexdens rubropilosa between 8:00 and 11:00 h and between 15:00 and 18:00 h. To investigate the attacking behaviour against the ants, flies were released individually inside an observation chamber containing a single leaf-cutting ant worker. Each parasitoid was confronted successively with a worker ant of A. sexdens rubropilosa, Atta laevigata Smith, Acromyrmex crassispinus Forel and Acromyrmex subterraneus molestans Santschi. Phorids of three species were identified: Neodohrniphora elongata Brown, Neodohrniphora declinata Borgmeier and Neodohrniphora tonhascai Brown. The three phorid species were active throughout the year and often along the same foraging trails, but N. elongata was the most frequent species. In the laboratory assay, N. elongata, N. declinata and N. tonhascai attacked workers of A. sexdens rubropilosa, A. laevigata and A. crassispinus, but not of A. subterraneus molestans.     

Electrophysiological responses of Atta sexdens rubropilosa workers to essential oils of eucalyptus and its chemical composition

The leaf-cutting ant Atta sexdens rubropilosa Forel, 1908 is the most harmful of the Eucalyptus pests, causing severe losses in wood production through defoliation. Various strategies have been tried and effort spent on the development of methods to control this pest, however no practical and environmentally acceptable one currently exists. In this work the chemical composition of the essential oil of seven Eucalyptus species was identified and the selectivity and sensitivity of antennal receptors of A. sexdens rubropilosa workers to the volatile compounds were determined using the electroantennographic technique (EAG and GC-EAD). Analysis by GC-EAD showed in E. cloesiana and E. maculata, respectively, seventeen and sixteen terpenes that elicited responses in ant workers' antennae, indicating the potential role of the essential oils as allelochemicals that determine the choice of the foraging material.     

Phenotypic plasticity in number of glomeruli and sensory innervation of the antennal lobe in leaf‐cutting ant workers (A. vollenweideri)

In the leaf‐cutting ant Atta vollenweideri, the worker caste exhibits a pronounced size‐polymorphism, and division of labor is dependent on worker size (alloethism). Behavior is largely guided by olfaction, and the olfactory system is highly developed. In a recent study, two different phenotypes of the antennal lobe of Atta vollenweideri workers were found: MG‐ and RG‐phenotype (with/without a macroglomerulus). Here we ask whether the glomerular numbers are related to worker size. We found that the antennal lobes of small workers contain ～390 glomeruli (low‐number; LN‐phenotype), and in large workers we found a substantially higher number of ～440 glomeruli (high‐number; HN‐phenotype). All LN‐phenotype workers and some small HN‐phenotype workers do not possess an MG (LN‐RG‐phenotype and HN‐RG‐phenotype), and the remaining majority of HN‐phenotype workers do possess an MG (HN‐MG‐phenotype). Using mass‐staining of antennal olfactory receptor neurons we found that the sensory tracts divide the antennal lobe into six clusters of glomeruli (T1–T6). In LN‐phenotype workers, ～50 glomeruli are missing in the T4‐cluster. Selective staining of single sensilla and their associated receptor neurons revealed that T4‐glomeruli are innervated by receptor neurons from the main type of olfactory sensilla, the Sensilla trichodea curvata. The other type of olfactory sensilla (Sensilla basiconica) exclusively innervates T6‐glomeruli. Quantitative analyses of differently sized workers revealed that the volume of T6 glomeruli scales with the power of 2.54 to the number of Sensilla basiconica. The results suggest that developmental plasticity leading to antennal‐lobe phenotypes promotes differences in olfactory‐guided behavior and may underlie task specialization within ant colonies. © 2009 Wiley Periodicals, Inc. Develop Neurobiol 70: 222–234, 2010.     

Toxic effects of Canavalia ensiformis L. (Leguminosae) on laboratory colonies of Atta sexdens L. (Hym., Formicidae)

Laboratory colonies of the leaf-cutting ants Atta sexdens that were fed daily with leaves of Canavalia ensiformis showed a high ant mortality, and a significant decrease in fungal garden volume, with complete depletion of nests after 11 weeks of treatment.     

Effects of leaves of Ipomoea batatas (Convolvulaceae) on nest development and on respiratory metabolism of leaf-cutting ants Atta sexdens L. (Hym., Formicidae)

Abstract: Laboratory colonies of the leaf-cutting ants Atta sexdens feed daily with leaves of Ipomoea batatas s howed ant mortality and a significant decrease in the size of the fungal garden after the second week, with complete depletion of nests after 5 weeks of treatment. The mean oxygen consumption rate of these ants was higher than the control (ants collected from nests feed with leaves of Eucalyptus alba ), suggesting a physiological action of the leaves of I. batatas on the ants in addition to the effect of inhibiting the growth of the fungal garden.     

Humidity preference for fungus culturing by workers of the leaf-cutting ant Atta sexdens rubropilosa

Summary: The hygropreference of gardening workers of the leaf-cutting ant Atta sexdens rubropilosa was investigated in the laboratory using a gradient of relative humidity. Gardening workers were placed, together with pieces of fungus garden, in small, interconnected nest chambers offering four different relative humidities: 33 %, 75 %, 84 % and 98 % RH. Workers were allowed to move freely between them and to relocate the fungus following their humidity preference. While workers distributed themselves randomly in the nest chambers, they located the fungus gardens in the chamber with the highest humidity. These results indicate that gardening workers are able to sense differences in relative humidity, and that this ability is shown when they are engaged in fungus culturing. Humidity is discussed as one of the relevant variables that probably underlay the evolution of regulatory responses for the control of fungus growth in leaf-cutting ants.     

Influence of leaf epicuticular waxes from cerrado species on substrate selection byAtta sexdens rubropilosa

The influence of leaf epicuticular waxes of nine woody species from cerrado (a savanna ecosystem of Brazil) on the foraging behavior of the leaf-cutting antAtta sexdens rubropilosa Forel, 1908 (Formicidae, Attini) was investigated. Some waxes showed neither inhibitory nor attractive effect over a series of tests, while a second group caused a delayed rejection. Waxes of two species were shown to be strongly deterrent to the ants. Since different waxes caused different responses, it is concluded that their chemical composition, rather than hydrophobicity or mechanical constraints, accounts for the observed effects. Plant species with leaves rejected by the ants did not necessarily have repellent waxes. On the other hand, no species with acceptable leaves yielded waxes with deterrent effects.     

Comparative analysis of fungal communities in colonies of two leaf-cutting ant species with different substratum preferences

Fungus gardens of leaf-cutting ants harbor diverse alien fungi in addition to their fungal cultivar. Previous work suggested that alien microorganisms are likely derived from the substrata foraged by ant workers and incorporated into the fungus gardens. To test this hypothesis, we sampled 1014 garden fragments from 16 field colonies of Atta sexdens rubropilosa (a dicot-cutting ant) and Atta capiguara (a grass-cutting ant) in Brazil. From a total of 615 fungal isolates recovered, we observed similar diversity of fungi between colonies of both ant species. However, fungal communities differed in composition of taxa between ant colonies. Trichoderma spirale, Trichosporon chiarellii and Penicillium citrinum were prevalent accounting for 18.5%, 12.2% and 11.7% of the total isolates, respectively. As expected, fungal communities clustered in two major groups supporting the hypothesis that plant substratum has an impact on the composition of the alien fungi found in leaf-cutting ant gardens.     

Behavioral and olfactory antennal responses of Solenopsis geminata (Fabricius) (Hymenoptera: Formicidae) workers to their dufour gland secretion

Behavioral and electrophysiological tests were performed to evaluate the responses of workers of the ant Solenopsis geminata (Fabricius) from different size categories to Dufour gland extracts. Morphometric measures based in head widths across eyes were used to determine worker sizes. Trail following response of different worker sizes to Dufour gland extract from workers of different sizes was assessed. For each worker size category olfactory responses to Dufour gland extracts were determined using electroantennography (EAG). Gas chromatography and mass spectrometry (GC-MS) were used to determine the chromatographic profile of Dufour gland secretion for each worker size. Morphometric measures permitted to classify the workers of S. geminata as large, medium and small workers. Medium S. geminata workers displayed a significantly higher behavioral response to Dufour gland extracts produced by medium size workers. Similarly, medium workers showed a significantly higher EAG response to Dufour gland extracts produced by medium sized workers. Chromatographic profile of Dufour gland secretions produced by workers showed that each size category exhibited a characteristic profile of the three main components considered as potential trail pheromone constituents. This work showed that medium workers of S. geminata exhibited a high trail-following behavior as well as a high antennal response to Dufour gland secretion. This and their relative abundance in field foraging areas, suggest that medium-sized workers are specialized in foraging activities.     

Response of workers of Atta sexdens rubropilosa (Hymenoptera: Formicidae) to mandibular gland compounds of virgin males and females

Abstract.  The secretion from the mandibular glands of males of the leaf-cutting ant Atta sexdens rubropilosa is responsible for the reaction of workers outside the nest at the time of sexual swarming. Workers respond with excitability and aggression when presented with the natural mixture of 4-methyl-3-heptanol and 4-methyl-3-heptanone, which is contained in the secretion of the male mandibular glands. Workers respond quickly to fractional amounts of one male equivalent. 4-Methyl-3-heptanone, from the virgin female mandibular glands causes much less response in workers, whereas an equimolar mixture of male and female pheromones gives a still less clear response. The male pheromone plays the most important part in the communication of workers outside the nest at this time.     

Odour learning and decision-making during food collection in the leaf-cutting antAcromyrmex lundi

Summary Recruited workers of the leaf-cutting antAcromyrmex lundi learn the odour of the food fragment initially carried by a successful scout, and use this cue as a decision criterion during food collection. Citral, a natural deterrent odour, can also be associated as a food signal, showing how powerful olfactory learning is in this species. The role of both food-odour learning and a previously reported leaf-marking pheromone during diet selection by leaf-cutting ants is discussed.     

Fluid intake rates in ants correlate with their feeding habits

This study investigates the techniques of nectar feeding in 11 different ant species, and quantitatively compares fluid intake rates over a wide range of nectar concentrations in four species that largely differ in their feeding habits. Ants were observed to employ two different techniques for liquid food intake, in which the glossa works either as a passive duct-like structure (sucking), or as an up- and downwards moving shovel (licking). The technique employed for collecting fluids at ad libitum food sources was observed to be species-specific and to correlate with the presence or absence of a well-developed crop in the species under scrutiny. Workers of ponerine ants licked fluid food during foraging and transported it as a droplet between their mandibles, whereas workers of species belonging to phylogenetically more advanced subfamilies, with a crop capable of storing liquids, sucked the fluid food, such as formicine ants of the genus Camponotus. In order to evaluate the performance of fluid collection during foraging, intake rates for sucrose solutions of different concentrations were measured in four ant species that differ in their foraging ecology. Scaling functions between fluid intake rates and ant size were first established for the polymorphic species, so as to compare ants of different size across species. Results showed that fluid intake rate depended, as expected and previously reported in the literature, on sugar concentration and the associated fluid viscosity. It also depended on both the species-specific feeding technique and the extent of specialization on foraging on liquid food. For similarly-sized ants, workers of two nectar-feeding ant species, Camponotus rufipes (Formicinae) and Pachycondyla villosa (Ponerinae), collected fluids with the highest intake rates, while workers of the leaf-cutting ant Atta sexdens (Myrmicinae) and a predatory ant from the Rhytidoponera impressa-complex (Ponerinae) did so with the lowest rate. Calculating the energy intake rates in mg sucrose per unit time, licking was shown to be a more advantageous technique at higher sugar concentrations than sucking, whereas sucking provided a higher energy intake rate at lower sugar concentrations.     

Life-history traits and parasitism rates of four phorid species (Diptera: Phoridae), parasitoids of Atta vollenweideri (Hymenoptera: Formicidae) in Argentina

Leaf-cutting ants in the genus Atta F. (Formicidae, Attini) are among the most important pest arthropods in Central and South America, consuming more vegetation than any other animal group. Among the organisms attacking ants in nature, flies of the family Phoridae have been proposed as the most promising biocontrol agents for pest ants. Four phorid species, Apocephalus setitarsus Brown, Myrmosicarius brandaoi Disney, Myrmosicarius gonzalezae Disney, and Eibesfeldtphora trilobata Disney, were reared from ants collected at Atta vollenweideri Forel nests and off foraging trails in Santa Fe province in Argentina. E. trilobata attacked larger ants and had bigger adults than the other species, also exhibiting the longest developmental time. Correlations between size of hosts and size of adults, as well as between size of adults and developmental times, could be established only in some cases. No differences were found between the sizes of the hosts from which males and females emerged. The natural percentage of parasitism varied throughout the seasons and seemed to be influenced by the extreme drought affecting the study site. We discuss why all four species would be suitable candidates for integrating an assemblage of biocontrol agents against A. vollenweideri.     

Spatial and Temporal Variation of Plant Fragment Removal by Two Species of <Emphasis Type="Italic">Atta</Emphasis> Leaf-Cutting Ants

Leaf-cutting ants are responsible for much of living plant fragment removal in Neotropical regions, causing major damage in both in natural and agricultural systems. To understand the variation in plant fragment removal by leaf-cutting ants and the influence that temporal and spatial factors have on such variation, we evaluated plant fragments collected by two ant species, Atta laevigata and A. sexdens rubropilosa in different seasons (rainy and dry) and in different forest systems (natural area of cerrado sensu stricto and monoculture of Eucalyptus sp.) in 2014 in Goianésia, Goiás, Brazil. Heavier plant fragments were removed during the dry season and in monoculture areas; A. laevigata removed heavier fragments than A. sexdens rubropilosa. Furthermore, we observed a positive association between the weight of collected fragments and the weight of worker ants and higher weights in A. laevigata. The study shows that the weight of plant fragments removed by leaf-cutting ants is conditioned temporally and spatially, and by the ant species involved. More detailed studies of each of these factors are need to better understand the dynamics of these foraging ants and how abiotic and biotic factors affect this process.     

The use of alarm pheromones to enhance bait harvest by grass-cutting ants

The enhancement of bait for the control of grass-cutting ants was investigated using two species of grass-cutting ant, Atta bisphaerica (Forel) and Atta capiguara (Gon?alves) (Hymenoptera: Formicidae). Bait was applied in loose piles to obtain a direct relationship between ant attraction and bait harvest. Enhancement with alarm pheromone compounds significantly increased the attractiveness and harvest of bait under certain conditions. A large proportion of the ants attracted to the enhanced bait were minor workers. These ants rarely transport bait because of their small size, and so it may be possible to increase the effect of bait enhancement by using smaller bait granules. Foragers of A. capiguara were less inclined to transport citrus-pulp bait than were those of Atta laevigata (Fr. Smith), a species that also harvests dicotyledonous plants. This emphasizes the importance of developing a bait matrix that is more acceptable to grass-cutting species. Nevertheless, the results suggest that alarm pheromone compounds have significant potential to improve the efficacy of baits for the control of grass-cutting ants.     

Methyl 4-methylpyrrole-2-carboxylate: a volatile trail pheromone from the leaf-cutting ant, tatta cephalotes

Methyl 4-methylpyrrole-2-carboxylate, a volatile trail pheromone previously identified from Atta texana, has been isolated and identified from the ant species, Atta cephalotes. The natural and the synthetic compound produce strong trail-following activity in a laboratory colony of A. cephalotes.     

Forager size of the leaf-cutting ant Atta sexdens (Hymenoptera: Formicidae) in a mature eucalyptus forest in Brazil

We determined the size characteristics of foragers of the leaf-cutting ant Atta sexdens in a mature eucalyptus forest in Campos dos Goytacazes, Rio de Janeiro State, Brazil, at daytime (7:30 to 10:00 hr) and nighttime (19:00 to 23:00 hr). There were no significant differences between daytime and nighttime ant mass (Ma), but leaf fragment mass (Ml) and burden (B = [Ma + Ml]/Ma), which indicates relative load capacity, were significantly greater at daytime. There was a positive linear relationship between Ma and Ml for the combined daytime and nighttime data, and increases in Ma resulted in lower B. We compared A. sexdens characteristics with published results for Atta cephalotes, a closely related species. A. sexdens is larger and therefore able to carry heavier loads, but its burden is about 72% of the average value for A. cephalotes. We suggest that the lower load capacity of A. sexdens in comparison to A. cephalotes is related to its relatively larger size.