Three-dimensional Reconstruction of Ovaries of Leaf-cutting Ant (Atta sexdens rubropilosa) Queens (Hymenoptera: Formicidae)

In this study, the ovary morphology of newly emerged ant queens of Atta sexdens rubropilosa was studied in whole mount preparations by confocal microscopy. The ovaries are composed of approximately 40 ovarioles, showing non-synchronic oocyte maturation. The terminal filament with clusters of undifferentiated cells was found at the distal end of the ovarioles. Next to this region is the germarium, composed of several elongated cystocytes interconnected by cytoplasmic bridges. The nurse cells (23-28 cells) result from asymmetric mitosis. Cytoskeleton analysis showed F-actin concentrated at the muscle cells of the external tunica and in fusomes inside the ovarioles. Microtubules were concentrated around the nuclei of the nurse and follicular cells. In contrast, the oocytes and the external tunica showed faint staining for tubulin.

Antimicrobial activity in the tick Rhipicephalus (Boophilus) microplus eggs: Cellular localization and temporal expression of microplusin during oogenesis and embryogenesis

Arthropods display different mechanisms to protect themselves against infections, among which antimicrobial peptides (AMPs) play an important role, acting directly against invader pathogens. We have detected several factors with inhibitory activity against Candida albicans and Micrococcus luteus on the surface and in homogenate of eggs of the tick Rhipicephalus (Boophilus) microplus. One of the anti-M. luteus factors of the egg homogenate was isolated to homogeneity. Analysis by electrospray mass spectrometry (ESI-MS) revealed that it corresponds to microplusin, an AMP previously isolated from the cell-free hemolymph of X (B.) microplus. Reverse transcription (RT) quantitative polymerase chain reactions (qPCR) showed that the levels of microplusin mRNA gradually increase along ovary development, reaching an impressive highest value three days after the adult females have dropped from the calf and start oviposition. Interestingly, the level of microplusin mRNA is very low in recently laid eggs. An enhance of microplusin gene expression in eggs is observed only nine days after the onset of oviposition, achieving the highest level just before the larva hatching, when the level of expression decreases once again. Fluorescence microscopy analysis using an anti-microplusin serum revealed that microplusin is present among yolk granules of oocytes as well as in the connecting tube of ovaries. These results, together to our previous data. suggest that microplusin may be involved not only in protection of adult female hemocele, but also in protection of the female reproductive tract and embryos, what points this AMP as a considerable target for development of new methods to control R. (B.) microplus as well as the vector-borne pathogens. (c) 2009 Elsevier Ltd. All rights reserved.

Worker laying in leafcutter ant Acromyrmex subterraneus brunneus (Formicidae, Attini)

We studied the process of offspring production in queenless colonies of Acromyrmex subterraneus brunneus, and particularly evaluated the ovary development of workers as a function of their age. For this, subcolonies were set up and evaluated at different periods of isolation from the queen (2, 4 and 6 months), besides individually labeled age groups. The subcolonies were assessed according to offspring production and ovaries containing oocytes or not. The evaluations showed worker oviposition and development of males originating from worker-laid eggs. At 2 months'absence of the queen, eggs and larvae were found, with eggs in a higher proportion than larvae. After 4 months, the proportion of eggs had reduced while larvae had increased, and a pupa was found in one subcolony. At 6 months, besides a higher share of larvae, one pupa and one adult male were found. Dissection of workers revealed ovaries containing oocytes during the periods of evaluation. Only a group of medium-sized and large workers, 23.3%, 20.9% and 37.5% of the population from each period assessed in queenless subcolonies respectively, presented developed oocytes in the ovary. The same was observed in colonies with a queen, with 17.6%, 19.6% and 7.8% of the group of dissected workers from each time period, respectively. With respect to worker age, we observed by dissection of the ovary, that the greatest percentage of individuals with ovarioles containing oocytes occurred at 45 days (6 weeks) up to 90 days (12 weeks). These results probably are associated with the workers reproduction and the laying of trophic and reproductive eggs in colonies with and without a queen; these eggs have distinct functions in each situation.

Variations in maternal care alter corticosterone and 17beta-estradiol levels, estrous cycle and folliculogenesis and stimulate the expression of estrogen receptors alpha and beta in the ovaries of UCh rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Germ line Cysts and the Formation of the Germinal Epithelium During the Female Gonadal Morphogenesis in Cyprinus Carpio (Teleostei: Ostariophysi: Cypriniformes)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reproductive pattern of the South American endemic shrimp Artemesia longinaris (Decapoda : Penaeoidea), off São Paulo State, Brazil

Sexual maturity, temporal reproductive pattern, and recruitment of juveniles were examined for the penaeoid shrimp Artemesia longinaris sampled for five and a half years in a tropical locality off the coast of São Paulo (23 degrees S), Brazil. Monthly samples were taken from January 1998 to June 2003 at depths between 5 and 45 m. Ovarian maturity was used to examine breeding in adult females. Recruitment was defined as the percentage of juveniles of the total number of individuals in each month and season. A total of 10 288 females and 5 551 males were collected. Estimated sizes (carapace length) at the onset of sexual maturity were 11.0 mm and 13.4 mm for males and females, respectively. Over the five and a half years, females with ripe gonads were found in every season, with the highest percentages in summer (January-March). Juvenile shrimps occur-red year-round. These results suggest a continuous reproduction of A. longinaris with temperature acting as an environmental stimulus for the duration of the ovary development cycle. These data and the hypothesis of the intrusion of the South Atlantic Central Water mass, which lowered water temperature and raised plankton production, suggest that the end of spring and the beginning of summer were the principal reproductive months. The classical paradigm of continuous reproduction at lower latitudes, with increased seasonality of breeding period at higher latitudes seems to apply to this species.

Reproduction and recruitment of the South American red shrimp, Pleoticus muelleri (Crustacea : Solenoceridae), from the Southeastern coast of Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Secretory cycle of the Dufour's gland in workers of the bumble bee Bombus terrestris L-(Hymenoptera : Apidae, Bombini)

The Dufour's gland of Bombus terrestris workers, of different ages and with varying degrees of ovary development, was studied with the aim to verify its involvement in reproduction. Measurements of the diameter and the length of the gland were made using an ocular micrometer adapted to a microscope. Transmission electron microscopy was used to study the morphology of the secretory cells and to analyze the secretory cycle. The glandular cells were considered to be near type II cells of NOIROT and QUENNEDEY (1974), a type that has not been described before in Hymenopterans. The results show that there is a correlation between the degree of ovary development and Dufour's gland activity of workers. The diameter of the gland and the secretory cell activity increased with increasing oocyte size in the ovary. Regressive conditions of the gland were observed, which are probably related to increasing worker age. To elucidate the production and releasing process of the secretion and to establish its precise function, a comparative analysis of the secretion process of the Dufour's gland of queens and workers is needed.

Ovarian ultrastructure in virgin queens of Apis mellifera L. narcotized by CO2

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morphological caste differences in the neotropical swarm-founding polistine wasp, Brachygastra lecheguana (Hymenoptera : Vespidae, Polistinae, Epiponini)

Morphological caste differences and ovary conditions were analyzed in colony of Brachygastra lecheguana. The results indicate the presence of individuals with ovary development but uninseminated (intermediates). In addition, these females were similar in morphology to workers, suggesting that, in this case, intermediates are ovary-developed workers. Queens were morphologically larger than workers and intermediates in some characters and smaller in others. Multivariate analysis showed that queens were highly distinct. These results indicate that castes are pre-imaginally determined in B. lecheguana.

An adaptive view of caste differentiation in the neotropical wasp Polybia (Trichothorax) sericea Olivier (Hymenoptera: Vespidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Non-allometric caste differences in Agelaia vicina (Hymenoptera, Vespidae, Epiponini)

Non-allometric and clear-cut caste dimorphism were found in two colonies of the social wasp Agelaia vicina. Such type of allometry was previously recorded in the Epiponini and its origin is pre-imaginally determined as in the other analysed species which present caste dimorphism. Multivariate analyses evidenced that the most intercastes discriminant characters were found in the metasoma and mesosoma while head differences were slight. Although queens were characterized by marked ovary development and insemination, the presence of some uninseminated queens was detected in one of the analysed colonies.

Cell death in ovarioles causes permanent sterility in Frieseomelitta varia worker bees

Frieseomelitta varia worker bees do not lay eggs even when living in queenless colonies, a condition that favors ovary development and oviposition in the majority of highly social bees. The permanent sterility of these worker bees was initially attributed to a failure in ovary morphogenesis and differentiation. Using transmission electron microscopy we found that at the beginning of the pupal phase the ovaries of F. varia workers are formed by four ovarioles, each of them composed of 1) a terminal filament at the apex of the ovarioles, containing juxtaposed and irregularly shaped cells, 2) a germarium with clusters of cystocytes and prefollicular cells showing long cytoplasmic projections that envelop the cystocyte clusters, 3) fusiform interfollicular and basal stalk precursor cells, and 4) globular, irregularly contoured basal cells with large nuclei. However, during the pupal phase an accentuated and progressive process of cell death takes place in the ovarioles. The dying cells are characterized by large membrane bodies, electron-dense apoptotic bodies, vacuoles, vesiculation, secondary lysosomes, enlarged rough endoplasmic reticulum cisternae, swollen mitochondria, pycnotic nuclei, masses of chromatin adjacent to the convoluted nuclear envelope, and nucleoli showing signs of fragmentation. Cell death continues in ovarioles even after the emergence of the workers. Once they become nurse bees, the ovaries have become transformed into a cell mass in which structurally organized ovarioles can no longer be identified. In F. varia workers, ovariole cell death most certainly is part of the program of caste differentiation.

Influence of the colony cycle on physiological and morphological caste variation in the perennial neotropical swarm-founding social wasp, Protonectarina sylveirae (Hymenoptera, Vespidae, Epiponini)

This paper describes the influence of the colony cycle on caste differences in Protonectarina sylveirae. Despite invariably keeping the main aspects of the conspicuous dimorphism between the castes, it is shown that: 1- queen-worker differences varied according to the progress of the colony cycle, 2- queens in pre-emergence colonies were smaller than those in male-producing ones, 3- workers were larger in pre-emergence than in other colony stages, 4- at particular steps of the colony cycle, non-inseminated ovary-developed queens and workers were detected. The size of the spermatheca differed between inseminated and non-inseminated queens with the highest values appearing in those bearing the most developed kind of ovary development. Differences between inseminated and non-inseminated queens were found in tergites III and IV, ovarian development and amount of fatty tissues. Because tergite size relates to gaster size, this character may be an important stimulus for selection of larger queens in the course of the colony cycle.

Nutritional and social hierarchy establishment of the primitively eusocial wasp Mischocyttarus cassununga (Hymenoptera, Vespidae, Mischocyttarini) and related aspects

The results on the nutritional influence in the social hierarchy establishment among females from six colonies of Mischocyttarus cassununga show that: - there was no caste determination in the pre-imaginal period concerning ingested food quantities, however, in relation to qualification (proteins and carbohydrate) the Mann-Whitney test applied showed variation among colonies: in two colonics (colony 3, p=0.0209; colony 4, p=0.0090) the larvae which ingested a higher quantity of solid food became adults of hierarchically higher position, however in two other colonies, colony 1 (p=0.6744) and colony 2 (p=0.3367) this difference was not statistically significant; in two experimentally manipulated colonies the caste determination was observed in the imaginal phase (it was verified that the larger the food quantity consumed, the higher the assumed hierarchical position by the individuals); - adult individuals of hierarchically higher position in two colonies ingested a greater amount of solid than liquid food (Mann-Whitney; p ≤ 0.05), however in the other three colonies there was no difference. It can be seen that for all colonies (2, 3, 4, 5 and 6) the high tendency to solid food ingestion occurs. In colony 1, this data was not collected because the observation purpose was different); - inside the colony hierarchy, the older individuals were the ones that occupied higher hierarchical positions; - more than one inseminated female per colony was found in the two experimentally manipulated colonies ; - ovarian development is greater in inseminated females; - the average height of the 3rd tergite was greater for inseminated females, indicating an adaptation to accommodate larger ovary development (t-Student test; p ≤ 0.05).