Transovarian Transmission of Blochmannia and Wolbachia Endosymbionts in the Neotropical Weaver Ant Camponotus textor (Hymenoptera, Formicidae).

Camponotus is a hyper-diverse ant genus that is associated with the obligate endosymbiont Blochmannia, and often also with Wolbachia, but morphological studies on the location of these bacteria in the queen's ovaries during oogenesis remain limited. In the present study, we used the Neotropical weaver ant Camponotus textor to characterize the ovary using histology (HE) techniques, and to document the location of Blochmannia and Wolbachia during oogenesis through fluorescence in situ hybridization (FISH). This is the first morphological report of these two bacteria in the same host with polytrophic meroistic ovaries and reveals that Blochmannia is found inside late-stage oocytes and Wolbachia is associated with the nuclei of the nurse cells. Our results provide insights into the developmental sequence of when these bacteria reach the egg, with Blochmannia establishing itself in the egg first, and Wolbachia only reaching the egg shortly before completing egg development. Studies such as this provide understanding about the mechanisms and timing of the establishment of these endosymbionts in the host.

The Postpharyngeal Gland: Specialized Organ for Lipid Nutrition in Leaf-Cutting Ants.

There are several hypotheses about the possible functions of the postpharyngeal gland (PPG) in ants. The proposed functions include roles as cephalic or gastric caeca and diverticulum of the digestive tract, mixing of hydrocarbons, nestmate recognition, feeding larvae, and the accumulation of lipids inside this gland, whose origin is contradictory. The current study aimed to investigate the functions of these glands by examining the protein expression profile of the PPGs of Atta sexdens rubropilosa (Hymenoptera, Formicidae). Mated females received lipid supplementation and their glands were extracted and analyzed using a proteomic approach. The protocol used combined two-dimensional electrophoresis and shotgun strategies, followed by mass spectrometry. We also detected lipid ß-oxidation by immunofluorescent marking of acyl-CoA dehydrogenase. Supplying ants with lipids elicited responses in the glandular cells of the PPG; these included increased expression of proteins related to defense mechanisms and signal transduction and reorganization of the cytoskeleton due to cell expansion. In addition, some proteins in PPG were overexpressed, especially those involved in lipid and energy metabolism. Part of the lipids may be reduced, used for the synthesis of fatty alcohol, transported to the hemolymph, or may be used as substrate for the synthesis of acetyl-CoA, which is oxidized to form molecules that drive oxidative phosphorylation and produce energy for cellular metabolic processes. These findings suggest that this organ is specialized for lipid nutrition of adult leaf-cutting ants and characterized like a of diverticulum foregut, with the ability to absorb, store, metabolize, and mobilize lipids to the hemolymph. However, we do not rule out that the PPG may have other functions in other species of ants.

Comparative morpho-physiology of the metapleural glands of two Atta leaf-cutting ant queens nesting in clayish and organic soils.

Queens of leaf-cutting ants found their nests singly, each consisting of a vertical tunnel and a final horizontal chamber. Because of the claustral mode of nest founding, the queen and/or her initial fungus garden are exposed to threats imposed by several soil pathogens, and the antibiotic secretions produced by their metapleural glands are considered a main adaptation to deal with them. Nests of two Atta leaf-cutting ant species, Atta vollenweideri and Atta sexdens rubropilosa, occur in different soil types, alfisols and oxisols. Their queens are known to excavate the initial nest in different soil horizons, clayish and organic, respectively, which differ in their fertility and associated microbiota. The aim of the present study was to comparatively investigate the morpho-physiology of the metapleural glands in queens of A. vollenweideri and A. sexdens rubropilosa, addressing the question whether the distinct selective pressure imposed by the microbiota in the two different soil types led to morpho-physiological differences in the metapleural glands that were consistent with their antiseptic function. The results revealed that metapleural glands of A. sexdens rubropilosa have a larger number of secretory cells, and consequently a higher production of antibiotic secretions, which may have been selected to allow nest founding at the superficial horizon of oxisols rich in organic matter and microorganisms. Glands of A. vollenweideri, on the contrary, presented fewer secretory cells, suggesting less production of antibiotic secretions. We argue that the excavation of deep founding nests in A. vollenweideri was primarily selected for during evolution to avoid the risk posed by flooding, and further hypothesize that a reduced number of cells in their metapleural glands occurred because of a weak pathogen-driven selective pressure at the preferred soil depth.

Mitochondrial and peroxisomal population in post-pharyngeal glands of leaf-cutting ants after lipid supplementation.

The post-pharyngeal gland (PPG) occurs in ants and some Sphecidae wasps. Among its several roles is the storage of lipids from food. In order to investigate the effect of lipids on the cell, especially on mitochondria and peroxisomes, the present study was aimed at examining the peroxisomal and mitochondrial population in the PPG of Atta sexdens rubropilosa after lipid supplementation by confocal laser scanning microscopy and transmission electron microscopy. Soybean oil provided as lipid supplement was not toxic for A. sexdens rubropilosa workers for the first 48 h and 120 h. However, the ultrastructural cytochemical analysis revealed an accumulation of lipid droplets in the PPGs of ants after lipid supplementation at 48 h and 120 h, and smaller lipid droplets in the basal membrane of the PPG epithelium, showing lipid mobilization from the PPG to the hemolymph. The lipid supplementation reduces the life expectancy of medium workers, probably due to the high lipid metabolism. Most importantly, the PPGs of medium workers of leaf-cutting A. sexdens rubropilosa is probably a specialized gland in the lipid metabolism, due to the increased mitochondrial and peroxisomal population inside cells after lipid supplementation; participation of peroxisomal population in the ß-oxidation of long chain fatty acids into smaller chains and participation of mitochondrial population in the ß-oxidation of fatty acids for energy, or mobilization of lipid derivatives from the PPG to hemolymph, a process that requires energy. However, the hypothesis that the PPGs convert lipids from food in aldehydes and/or hydrocarbons must be better investigated.

Morphophysiological differences between the metapleural glands of fungus-growing and non-fungus-growing ants (Hymenoptera, Formicidae).

The metapleural gland is an organ exclusive to ants. Its main role is to produce secretions that inhibit the proliferation of different types of pathogens. The aim of the present study was to examine the morphophysiological differences between the metapleural gland of 3 non-fungus-growing ants of the tribes Ectatommini, Myrmicini, and Blepharidattini and that of 5 fungus-growing ants from 2 basal and 3 derived attine genera. The metapleural gland of the non-fungus-growing ants and the basal attine ants has fewer secretory cells than that of the derived attine ants (leaf-cutting ants). In addition, the metapleural gland of the latter had more clusters of secretory cells and sieve plates, indicating a greater storage capacity and demand for secretion in these more advanced farming ants. The glands of the derived attine ants also produced higher levels of polysaccharides and acidic lipids than those of Myrmicini, Blepharidattini, and basal attines. Our results confirm morphophysiological differences between the metapleural glands of the derived attines and those of the basal attines and non-fungus-growing ants, suggesting that the metapleural glands of the derived attines (leaf-cutting ants) are more developed in morphology and physiology, with enhanced secretion production (acidic lipids and protein) to protect against the proliferation of unwanted fungi and bacteria in the fungal garden, it is possible that leaf-cutting ants may have evolved more developed metapleural glands in response to stronger pressure from parasites.

Secretory profile of metapleural gland cells of the leaf-cutting ant Acromyrmex coronatus (Formicidae: Attini).

Ants present a pair of metapleural glands located at the posterolateral end of the thorax. Because of its importance in the social organization of ants, the present study was aimed at describing the morphophysiology of this gland in three worker castes of Acromyrmex coronatus, focused on secretory activity using histological and histochemical techniques. Our findings revealed that the secretory and the storage portions of this gland are connected by extracytoplasmic portion of canaliculi that drain the secretion from each secretory cell to the collecting chamber. This secretion contains glycoproteins. In minor workers, the secretion contains higher levels of polysaccharides when compared to that of major workers, supporting the role of the metapleural gland in the maintenance of the fungus garden. The nucleus as well as cytoplasm of secretory cells were strongly positive for RNA indicating that these cells are active in the synthesis of proteins and lipids, compounds found in the final secretion. The variant of the CEC revealed that the secretory activity of the entire gland is synchronous, as all cells exhibit the result.

The functional morphology of the metapleural gland of the leaf-cutting ant Atta laevigata (Formicidae: Attini).

Given the importance of the metapleural gland in ants, the present study aimed at describing it in minor, media, and major workers of Atta laevigata, focusing their physiology using scanning electron microscopy (SEM), histologic, and histochemical techniques. The results revealed that this gland is paired and located in the metathoracic region, consisted of two portions: secretory and the storage ones. Both portions are connected by extracytoplasmic canaliculi that drain the secretion of each secretory cell they form groups that open into the perforated plate located in the wall of the collecting chamber. The oval-shaped and elongated secretory cells form clusters of approximately 20 cells, bundled together by connective tissue. Each secretory cell connects to the reservoir through a canaliculus divided into an intra- and extracytoplasmic portion. The collecting chamber and the reservoir are internally lined by a single squamous epithelium with a cuticular intima, with folds in the collecting chamber and smooth in the remaining of the reservoir. External muscle layers surrounding the reservoir are observed which aid the release of secretion by the gland opening (one on each side) located in the exoskeleton near coxae of hind legs. Secretory cells were strongly positive for acidic and total lipids, proteins, and acidic polysaccharides, suggesting the glycolipoproteins nature of the final secretion. Morphometrically, the diameter of the reservoir is proportional to body size of minor, media, and major workers. This implies a larger capacity to produce secretion, including antibiotic substances, by the individuals that perform specific tasks within the colony.

Modelo arquitetônico de ninhos da formiga Ectatomma vizottoi Almeida (Hymenoptera, Formicidae) / Nest architecture of the ant Ectatomma vizottoi Almeida (Hymenoptera, Formicidae)

Para descrever o padrão de construção e ocupação dos ninhos de Ectatomma vizottoi, foram escavados oito ninhos durante o período de novembro de 2004 a agosto de 2006, em Dourados-MS. Os orifícios de entrada ou saída dos ninhos desta espécie possuem formato elipsóide, com acesso a um túnel alargado, similar a uma antecâmara, a qual é conectada a câmaras mais profundas do ninho. Os ninhos podem alcançar até 360,0 cm de profundidade, e apresentam de três a dez câmaras. São construídos independentemente de raízes de plantas, e o solo de seus túneis e câmaras é compactado, apresentando compartimentos específicos para depósitos de detritos da colônia. O aumento da densidade de formigas conduz a um aumento do número de câmaras, bem como da profundidade do ninho, mantendo, entretanto, um volume médio para o mesmo, evidenciando um padrão para o tamanho das câmaras dos ninhos.

The construction pattern and occupancy of Ectatomma vizottoi was described, during November 2004-August 2006 period, in Dourados, state of Mato Grosso do Sul by excavation of eight nests. The entrance openings, or exit, have an ellipsoid shape and lead to a wide tunnel, similar to a hall, which is connected to other deeper chambers in the nest. The nests reach up to 360 cm deep and contain three to ten chambers. Nests are constructed independently of plants and roots presence, and the soil of the tunnels and chambers are compacted, with specific compartments for colony deposit of detritus. The increase of the ant density led to an increase in nest depth and number of chambers, maintaining an average nest volume, which indicates a standard size for nest chambers.