Morphological aspects of spermatogenic cells at different stages of sexual maturation in black isogenic C57BL/6/Uni mice

In order to study the morphological changes that occur in cells of the testes of isogenic black mouse C57BL/6/Uni into three periods during spermatogenetic used 15 mice divided into 3 groups of 5 animals with 40,50 and 60 days of age. The mice were sacrificed and weighed. Testicles were weighed and measured, and histologically processed and stained with HE, PAS and Masson Massom-H and evaluated under light microscopy. It was observed that group I with 40 days of age in the seminifcrous tubules had a lumen with sparse small amount of interstitial tubular cells. In the seminiferous epithelium type A spermatogonia, intermediate and B were identified, which occupied the compartment adbasal and intermingled with these cells in spermatocytes I in Pachytene and leptotene was observed, whereas in the adluminal compartment Golgi phase spermatids we observed the presence of acrosomal granule. In group II, the cells of the seminiferous epithelium were developed and it was observed in round spermatids cephalic hood phase plus many elongated spermatids in acrosome phase and Sertoli cells. In Group III, 60 days old, it was found that seminiferous epithelium which was of the tubules had elongated spermatids in acrosome phase and maturation, with elongated nuclei and acrosomal system typical of spermiation in the presence of sperm and residual bodies near the tubular lumen. Therefore morphological evolution of germ cell testicular spermatids can be checked and recognized in its four phases: Golgi, cap, acrosome and maturation over the age of the animal.

Genome-Wide Analysis of Differentially Expressed Genes During the Early Stages of Tomato Infection by a Potyvirus

Plant responses against pathogens cause up-and downward shifts in gene expression. To identify differentially expressed genes in a plant-virus interaction, susceptible tomato plants were inoculated with the potyvirus Pepper yellow mosaic virus (PepYMV) and a subtractive library was constructed from inoculated leaves at 72 h after inoculation. Several genes were identified as upregulated, including genes involved in plant defense responses (e. g., pathogenesis-related protein 5), regulation of the cell cycle (e. g., cytokinin-repressed proteins), signal transduction (e. g., CAX-interacting protein 4, SNF1 kinase), transcriptional regulators (e. g., WRKY and SCARECROW transcription factors), stress response proteins (e. g., Hsp90, DNA-J, 20S proteasome alpha subunit B, translationally controlled tumor protein), ubiquitins (e. g., polyubiquitin, ubiquitin activating enzyme 2), among others. Downregulated genes were also identified, which likewise display identity with genes involved in several metabolic pathways. Differential expression of selected genes was validated by macroarray analysis and quantitative real-time polymerase chain reaction. The possible roles played by some of these genes in the viral infection cycle are discussed.

Description of gamontogonic and sporogonic stages of Hepatozoon spp. (Apicomplexa, Hepatozoidae) from Caudisoma durissa terrifica (Serpentes, Viperidae)

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

Seasonal effect of three pasture plants species on the free-living stages of Haemonchus contortus

Avaliaram-se o desenvolvimento e a sobrevivência dos estágios de vida livre de Haemonchus contortus em fezes obtidas de ovinos infectados por esse nematódeo, depositadas nas quatro estações do ano, em pastagens de três espécies de gramíneas (Brachiaria decumbens cv Australiana, Cynodon dactylon cv. Coast-cross e Panicum maximum cv. Aruana), com duas alturas (5cm e 30cm) no momento da deposição das fezes. Amostras de fezes e de capim foram coletadas uma, duas, quatro, oito, 12 e 16 semanas após a deposição das fezes e processadas em laboratório, para recuperação de larvas infectantes. O maior número de larvas infectantes foi recuperado das amostras de capim e de fezes nas pastagens com 30cm de altura. As condições climáticas que mais favoreceram o desenvolvimento e a sobrevivência das larvas nas fezes, bem como a sobrevivência e a manutenção das larvas no capim foram aquelas com temperaturas médias em torno de 17ºC, acompanhadas de baixas precipitações pluviométricas. As temperaturas mais altas e as precipitações elevadas, que ocorrem na chamada estação das águas, foram desfavoráveis para a recuperação de larvas infectantes da pastagem. de forma geral, a pastagem de Aruana foi a que possibilitou as maiores recuperações de larvas.

Postlarval stages and growth patterns of the spider crab Pyromaia tuberculata (Brachyura, Majidae) from laboratory-reared material

The spider crab Pyromaia tuberculata was introduced into southeastern Brazil; ovigerous material was collected and reared in the laboratory. Morphologic changes and growth patterns of post-larval development are reported. Results show that within-stage size variation is lowest in mature stages, especially in the case of females in which there is an apparent size threshold for the last juvenile stages to undergo the puberty molt. A prepuberty molt taking place at the fourth crab stage is indicated by analyzing the allometric growth of the abdomen in females. In contrast, the same procedure using the allometric growth of chelae failed in detecting both the prepuberty and puberty molts in males. Conversely to females, which develop a complex brood chamber at the puberty molt, the enlargement of chelae was not consistent in all postpuberty males. The short instar sequence of this species, in no case exceeding nine stages, is marked by conspicuous morphologic alterations achieved at each molt. Almost all stages can be identified by examining diagnostic features of rostrum, abdomen, sternum, and pleopods.

Ecological distribution and reproductive period of the hermit crab Loxopagurus loxochelis (Anomura, Diogenidae) on the northern coast of São Paulo State, Brazil

The present study aimed to determine the ecological distribution of Loxopagurus loxochelis as a function of selected environmental factors, as well as its reproductive period based on the combined analysis of the presence of ovigerous females and the development of the gonad. The collections were carried out monthly from January to December 2000 in the region of Ubatuba (SP), using a fishing boat equipped with two double rig nets, in isobaths of 5, 10, 15, 20, 25, 30, 35 and 40 m, at which depths samples of sediment and water were collected. The cephalothoracic shield length and sex of the animals were determined; the abdomen was dissected to verify gonad development stages. A total of 334 individuals was obtained ( 196 males, 48 ovigerous females and 90 non-ovigerous females) with sizes varying from 3.0 to 8.6 mm (5.7 +/- 1.0 mm). A greater abundance of L. loxochelis (95.2%) was observed at depths of 20 and 25 m. These sites mainly revealed a substratum stable with a very fine sand fraction, which facilitates the habit of embedding shown by this hermit. With regard to gonadal analysis, it was possible to classify four gonad development stages for each sex: immature, rudimentary, developing and developed. It was observed that in the winter months about 71% of the females had gonads either developing or developed and 67% were ovigerous. Therefore, the winter period can be considered the peak of reproduction for this species, characterizing a typical seasonal-continuous reproduction.

Larval stages of Stenocionops furcatus (Olivier, 1791) (Decapoda : Brachyura : Majoidea) and a reappraisal of larval morphological characters for Mithracidae

Stenocionops furcatus is a spider crab found in the western Atlantic, from Georgia, USA to Rio Grande do Sul, Brazil, on sand, coral, rocks or mud bottoms from the intertidal zone to 180 m. We describe all laboratory-reared larval stages of S. furcatus obtained from the northern coast of São Paulo State, Brazil, and compare our data with existing larval descriptions for the genus and other mithracids. The larval development of S. furcatus consists of two zoeal stages and one megalopa. The durations of the first and second zoeal stage were similar to4 and 5 days respectively, the megalopa appearing 10-18 days after hatching. Our results show that the zoeae of S. furcatus differ from those of other Mithracidae by possessing four setae on the proximal lobe of the coxal endite of the maxilla, instead of five, and by the presence of mid-dorsal setae on the third abdominal somite in the second zoeal stage, which are lacking in other mithracids. Larval descriptions for Stenocionops in two previous publications were attributed to the subspecies S. furcatus coelatus from the Caribbean. Larvae from Brazilian waters closely resemble one of these accounts, suggesting that this taxon extends beyond the West Indies and that the other description represents larvae of S. furcatus furcatus. Additional morphological details, not available previously, are provided.

Larval stages of Paradasygyius depressus (Bell, 1835) (Crustacea : Decapoda : Brachyura : Majidae) and a phylogenetic analysis for 21 genera of Majidae

All larval stages and the first crab instar of Paradasygyius depressus (Bell) were obtained in laboratory culture. Larval development consists of two zoeal stages, followed by the megalopa. Each larval stage is described in detail. Beginning with the first zoea, the duration of each stage was 4--7 (4.5 +/- 0.7), 4-5 (4.5 +/- 0.5), and 7 days, the megalopa and first crab instar appearing 11 +/- 1 and 15 days after hatching, respectively. A phylogenetic analysis of 21 genera of Majidae is provided based on 34 zoeal and three megalopal characters. The phylogenetic analysis resulted in four equally parsimonious trees 173 steps long (CI = 0.66, RI = 0.71, and RC = 0.47) supporting the monophyly of Oregoniinae, Majinae, and Inachinae (with the exclusion of Macrocheira de Haan incertae sedis). Based on general agreement of sister-group hypotheses, we provide sets of larval characters that define Oregoniinae, Majinae, and Inachinae. Our phylogenetic hypothesis suggests that Oregoniinae is the most basal clade within the Majidae, and Majinae and the clade (Epialtus H. Milne Edwards + Inachinae [excluding Macrocheira incertae sedis]) are sister taxa. Within Inachinae, all trees suggest that Inachus Weber and Macropodia Leach are sister taxa nested as the most derived clade, followed by Achaeus Leach, Pyromaia Stimpson, Paradasygyius Garth, Anasimus A. Milne-Edwards, and the most basal Stenorhynchus Lamarck. The sister-group relationships of the clade (Pisa Leach (Taliepus A. Milne-Edwards + Libinia Leach)), Mithrax Latreille and Microphrys H. Milne Edwards remained unresolved.

On the larval stages of Macrocoeloma diplacanthum (Decapoda : Brachyura : Majidae), and a review of mithracine phylogenetic aspects

Larval development of Macrocoeloma diplacanthum (Stimpson) consists of two zoeal stages, followed by the megalopa. Each larval stage is described in detail. The duration of the zoeal stages was 2-3 (2.4 +/- 0.5) and 3-4 (3.5 +/- 0.5) days for the first and second zoea, respectively, the megalopa phase appearing 6-8 (7.0 +/- 0.5) days after hatching. Unlike for other majids, zoeal stages of M. diplacanthum can be readily distinguished by their distended forehead with strong underlying muscle bands, undercut dorsal carapace spine, and spine on the terminal endopod segment of the first maxilliped. No other known mithracine or majid zoeae exhibit this combination of features. Our zoeal account of M. diplacanthum from Mexico is remarkably consistent with Floridian specimens previously described. However, we have found some differences between descriptions, which could be attributed to natural variation or inadequate description. Previous attempts to evaluate the relationships within Mithacinae have been based on larval characters widely distributed throughout Majidae and therefore are considered inadequate to infer sister-group relationships. The phylogenetic analysis of majids suggested that the position of Mithracinae is still uncertain, as is its monophyletic status. We recommend that additional characters, particularly of the megalopa phase, be sought for a better resolution of majid evolutionary history.

Ultrastructure of the midgut epithelium of Meliponinae larvae with different developmental stages and diets

The comparative study of the ultrastructure of the midgut epithelium of stingless bee larvae that eat plant protein (pollen) and animal protein (carrion) throughout the larval phase, shows variations in the digestive cells that are only relative to larval aging and not to the type of larval diet. The cells of older larvae present a cytoplasm with empty spaces that result from emptying of lipid and glycogen stocks, and the presence of autophagic vacuoles. These results are discussed in relation to the hypothesis that variations in the digestive tract of insects may be associated with different diets or phylogeny. We conclude that different diets do not determine cell morphology adaptations in the studied species. As the variations in the ultrastructure of the midgut epithelium are the same in all studied species, including the necrophagous species Trigona hypogea, throughout the larval stage, this sequence of changes seems to be due to different physiological state during larval development.

Ultrastructure features of the midgut of the female adult Amblyomma cajennense ticks Fabricius, 1787 (Acari: Ixodidae) in several feeding stages and subjected to three infestations

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

Morpho-histochemical characterization of salivary gland cells of males of the tick Rhipicephalus sanguineus (Acari: Ixodidae) at different feeding stages: description of new cell types

This study describes the changes undergone by cells of the salivary glands of unfed and feeding (at day two and four post-attachment) Rhipicephalus sanguineus males, as well as new cell types. In unfed males, types I and II acini are observed with cells undifferentiated, undefined 1 and 2 (the latter, with atypical granules), a, c1 and c3; type III is composed of cells d and e; and type IV present cells g. In males at day two post-attachment, type I acini exhibit the same morphology of unfed individuals. An increase in size is observed in types II, III, and IV, as cells are filled with secretion granules. Some granules are still undergoing maturation. In type II acinus, cells a, b and c1-c8 are observed. Cells c7 and c8 are described for the first time. Cells c7 are termed as such due to the addition of polysaccharides in the composition of the secretion granules (in unfed individuals, they are termed undefined 1). Type III acini exhibit cells d and e completely filled with granules, and in type IV, cells g contain granules in several stages of maturation. In males at day four post-attachment, type I acini do not exhibit changes. Granular acini exhibit cells with fewer secretion granules, which are already mature. In type II acini, cells a, b, c1-c5 are present, type III exhibit cells d and e, and type IV contain cells g with little or no secretion. This study shows that in the salivary glands of R. sanguineus males, cells a, c1, and c3 of type II acinus, and cells d and e of type III do not exhibit changes in granular content, remaining continuously active during the entire feeding period. This indicates that during the intervals among feeding stages, gland cells reacquire the same characteristics found in unfed individuals, suggesting that they undergo reprogramming to be active in the next cycle.

Evaluation of the antigenotoxicity of polysaccharides and beta-glucans from Agaricus blazei, a model study with the single cell gel electrophoresis/Hep G2 assay

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A morphological view of the relationship between indirect flight muscle maturation and the flying needs of two species of advanced eusocial bees

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

Histopathology and Ultrastructure Features of the Midgut of Adult Females of the Tick Amblyomma cajennense Fabricius, 1787 (Acari: Ixodidae) in Various Feeding Stages and Submitted to Three Infestations

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