Landscape genetics across the Andes Mountains: Environmental variation drives genetic divergence in the leaf-cutting ant Atta cephalotes

Disentangling the mechanisms underlying spatial distribution of genetic variation into those due to environment or physical barriers from mere geographic distance is challenging in complex landscapes. The Andean uplift represents one of the most heterogeneous habitats where these questions remain unexplored as multiple mechanisms might interact, confounding their relative roles. We explore this broad question in the leaf-cutting ant Atta cephalotes, a species distributed across the Andes Mountains using nuclear microsatellite markers and mtCOI gene sequences. We investigate spatial genetic divergence across the Western range of Northern Andes in Colombia by testing the relative role of alternative scenarios of population divergence, including geographic distance (IBD), climatic conditions (IBE), and physical barriers due to the Andes Mountains (IBB). Our results reveal substantial genetic differentiation among A. cephalotes populations for both types of markers, but only nuclear divergence followed a hierarchical pattern with multiple models of genetic divergence imposed by the Western range. Model selection showed that IBD, IBE (temperature and precipitation), and IBB (Andes mountains) models, often proposed as individual drivers of genetic divergence, interact, and explain up to 33% of genetic divergence in A. cephalotes. IBE models remained significant after accounting for IBD, suggesting that environmental factors play a more prominent role when compared to IBB. These factors in combination with idiosyncratic dispersal patterns of ants appear to determine hierarchical patterns of gene flow. This study enriches our understanding of the forces shaping population divergence in complex habitat landscapes.

Traditional knowledge regarding entomophagy in San Martín, Peruvian Amazon

The consumption of insects is a widespread practice among indigenous or native peoples of the Amazon. To assess the knowledge of the diversity of resources for entomophagy from the perspective of these peoples, testimonies or references about knowledge and feeding traditions of 100 people were collected in 37 localities in seven provinces of the department of San Martín in the basin of the Huallaga River. One cumulative species curve and the probability function of new species were estimated, then the probability of not finding a new species (99.5 %) to n100 was determined. The specimens that were captured in the field (54 %) were compared with representative specimens and databases, and the information provided by the participants was analysed to approximate the preliminary taxonomic locations of the remaining part of the sample. We found 46 resources for entomophagy and reported for first time in the Peruvian Amazon, the feeding with Chrysophora chrysochlora, Podalia sp., Lusura chera, and Cymothoidae, among others. Entomophagy is a deeply rooted practice in the native and riverine populations of the Huallaga basin, where Rhynchophorus palmarum, Rhinostomus barbirostris, Atta cephalotes sspp. and Brassolis sophorae were the most consumed for 78 % – 97 % of people. The least consumed species have the common characteristic of being scarce and they were part of the diet of the oldest segment in previous decades. At least 10 resources ceased to be consumed by the members of the sample. In addition to nutritional potential, the diversity of edible arthropod fauna represents complementary values for community health and cultural identity; however, most of these resources (87 %) are threatened and could disappear in brief time, as is the knowledge related to their use.

The role of the Andean uplift as an asymmetrical barrier to gene flow in the Neotropical leaf-cutting ant Atta cephalotes

Neotropical diversification by the Andean uplift is typically addressed on a large evolutionary scale (e.g. speciation), even though many species are still distributed in both sides of the mountains. The three parallel mountain ranges in the northern Andes (Colombia) impose a major geographical barrier to species' migration from South to Central America. How important these barriers are for conspecific diversification of cross-Andean species such as the leaf-cutting ants remains largely unknown. To answer this question, we studied the mtCOI gene of Atta cephalotes, the most widely distributed leaf-cutting ant species. Our hierarchical analyzes evidenced substantial genetic structure among regions and populations, suggesting a more complex biogeographical history of Andean populations than previously thought. These mountains seem to isolate Central American and Western Colombian populations from the rest of A. cephalotes in South America. Population and migration modelling are consistent with the origin of this species in South America and a major role of the Eastern cordillera as a geographical barrier to historical gene flow, restricting dispersion from north to south. These findings provide insights into the role of the Andean uplift as barrier to gene flow and, eventually, implications for monitoring and designing management strategies for leaf-cutting ants.

Immunity and survival response of Atta cephalotes (Hymenoptera: Myrmicinae) workers to Metarhizium anisopliae infection: Potential role of their associated microbiota

Leaf-cutting ants of the genera Atta and Acromyrmex are at constant risk of epizootics due to their dense living conditions and frequent social interactions between genetically related individuals. To help mitigate the risk of epizootics, these ants display individual and collective immune responses, including associations with symbiotic bacteria that can enhance their resistance to pathogenic infections. For example, Acromyrmex spp. harbor actinobacteria that control infection by Escovopsis in their fungal gardens. Although Atta spp. do not maintain symbiosis with protective actinobacteria, the evidence suggests that these insects are colonized by bacterial microbiota that may play a role in their defense against pathogens. The potential role of the bacterial microbiome of Atta workers in enhancing host immunity remains unexplored. We evaluated multiple parameters of the individual immunity of Atta cephalotes (Linnaeus, 1758) workers, including hemocyte count, encapsulation response, and the antimicrobial activity of the hemolymph in the presence or absence of bacterial microbiota. Experiments were performed on ants reared under standard conditions as well as on ants previously exposed to the entomopathogenic fungus Metharrizium anisopliae. Furthermore, the effects of the presence/absence of bacteria on the survival of workers exposed to M. anisopliae were evaluated. The bacterial microbiota associated with A. cephalotes workers does not modulate the number of hemocytes under control conditions or under conditions of exposure to the fungal pathogen. In addition, infection by M. anisopliae, but not microbiota, increases the encapsulation response. Similarly, the exposure of workers to this fungus led to increased hemolymph antimicrobial activity. Conversely, the removal of bacterial microbiota did not have a significant impact on the survival of workers with M. anisopliae. Our results suggest that the bacterial microbiota associated with the cuticle of A. cephalotes workers does not play a role as a modulator of innate immunity, either at baseline or after exposure to the entomopathogen M. anisopliae. Further, upon infection, workers rely on mechanisms of humoral immunity to respond to this threat. Overall, our findings indicate that the bacterial microbiota associated with A. cephalotes workers does not play a defensive role.

Occurrence of Leaf-Cutting and Grass-Cutting Ants of the Genus Atta (Hymenoptera: Formicidae) in Geographic Regions of Brazil

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.

Characterization of a New Set of Microsatellite Markers Suggests Polygyny and Polyandry in Atta cephalotes (Hymenoptera: Formicidae)

The leaf-cutting ant, Atta cephalotes L. (1758), is a major herbivore with great economic impact in the Neotropics. Because of its broad range and human-mediated dissemination, the ecology of this ant has received considerable attention; however, questions concerning its population genetics, dispersal, and social structure remain unexplored. Here, we aimed to identify and provide information on molecular and statistical performance of a suite of polymorphic microsatellite markers for A. cephalotes while demonstrating their utility for further genetic studies. We designed primer sequences targeting thousands of microsatellite loci and then screened 30 of these for amplification and polymorphism. Fifteen of these loci were selected and used to evaluate their polymorphism using 74 ants sampled from 15 different nests of the same location. This set of 15 loci exhibited variation of 2–20 alleles, with a mean heterozygosity of 0.57. All loci followed Hardy–Weinberg expectations with no evidence of linkage disequilibrium, while two loci showed evidence of null alleles. Our preliminary analysis suggested substantial nest differentiation with no population viscosity within the sampled location, as well as colonies with multiple queens (polygyny) and paternity (polyandry). Our newly identified microsatellites have proven to be highly informative to investigate gene flow, social structure and reproduction of this key agricultural pest.