Structural and ultrastructural characteristics of the yolk syncytial layer in Prochilodus lineatus (Valenciennes, 1836) (Teleostei; Prochilodontidae)

The yolk syncytial layer (YSL) has been regarded as one of the main obstacles for a successful cryopreservation of fish embryos. The purpose of this study was to identify and characterize the YSL in Prochilodus lineatus, a fish species found in southeastern Brazil and considered a very important fishery resource. Embryos were obtained through artificial breeding by hormonal induction. After fertilization, the eggs were incubated in vertical incubators with a controlled temperature (28 degrees C). Embryos were collected in several periods of development up to hatching and then fixed with 2% glutaralclehyde and 4% paraformaldehyde in 0.1 M sodium phosphate buffer (pH 7.3). Morphological analyses were carried out under either light, transmission or scanning electron microscopy. The formation of the YSL in P. lineatus embryos starts at the end of the cleavage stage (morula), mainly at the margin of the blastoderm, and develops along the embryo finally covering the entire yolk mass (late gastrula) and producing a distinct intermediate zone between the yolk and the endodermal cells. The YSL was characterized by the presence of microvilli on the contact region with the yolk endoderm. A cytoplasmic mass, full of mitochondria, vacuoles, ribosomes, endomembrane nets and euchromatic nuclei, indicated a high metabolic activity. This layer is shown as an interface between the yolk and the embryo cells that, besides sustaining and separating the yolk, acts as a structure that makes it available for the embryo. The structural analyses identified no possible barriers to cryoprotectant penetration.

Identity by descent and candidate gene mapping of Richieri-Costa and Pereira syndrome

The Richieri-Costa-Pereira syndrome is a rare autosomal recessive disorder characterized by short stature, Robin sequence, cleft mandible, pre/postaxial anomalies and clubfoot. of 15 families reported with this disorder 14 are from Brazil suggesting a founder effect. We studied 15 families using identity-by-descent as a hypothesis to attempt gene localization We have examined through linkage analysis 497 polymorphicmarkers and also performed direct sequencing of exons for 10 candidate genes selected on the basis of their expression in the developing mandible and limb. No evidence for allele sharing at any locus tested or mutations in candidate genes was found. Additional higher resolution mapping, new families and other candidate genes might improve future chances of gene identification. (C) 2003 Wiley-Liss, Inc.

Cryogenic preservation of embryos of Prochilodus lineatus (Valenciennes, 1836) (Characiforme; Prochilodontidae)

While the freezing techniques of mammal embryos have been providing promising results, the cryopreservation of teleostean eggs and embryos have remained unsuccessful up to now. Therefore, this work aimed to develop a procedure of cryogenic preservation of embryos of Prochilodus lineatus and to observe, at both structural and ultrastructural levels, the morphological alterations that took place after the application of freezing/thawing techniques. The embryos at the morula stage could not tolerate exposure to the cryoprotectants ethylene glycol monomethyl ether, propylene glycol monomethyl ether, methanol, dimethyl sulphoxide and propylene glycol, presenting 100% of mortality. Embryos at the 4- to 6-somites stage tolerated exposure to propylene glycol and dimethyl sulphoxide, and the results revealed no significant differences (alpha = 0.05) regarding survival from both treatments. None of the freezing, thawing and hydration protocols was effective on preserving embryo viability. The ultrastructural analyses of frozen and thawed embryos showed that cells from ectoderm, somites, notochord and endoderm were structurally intact, with well preserved nuclei and mitochondria. The yolk globules were able to tolerate the freezing process, but the yolk syncytial layer was unorganized, displaying an electron-dense and compacted appearance, collapsed reticules, nuclei with modified chromatin and ruptures on the plasmatic membrane at the contact zone with endoderm. It might be concluded that the procedures tested for freezing were unable to avoid the formation of intracellular ice crystals, leading to drastic morphological modifications and making P. lineatus embryos unviable.

Ultrastructural studies of the mandibular gland of Melipona quadrifasciata Lepeletier, 1836 (Apidae, Meliponini): a comparison between workers and queens

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

A comparative cytochemical study of the dufour gland in the eusocial bee Apis mellifera Linne, 1758 and Melipona bicolor Lepeletier, 1836

Dufour glands of Apis mellifera and Melipona bicolor were studied under light and transmission electron microscopy, using the cytochemical techniques of mercury bromophenol blue for protein detection, imidazole-buffered osmium tetroxide selective staining of unsaturated lipids, lanthanum nitrate for intercellular junction identification and zinc-iodide-osmium tetroxide for cytoplasmic endomembrane visualization. The results in both species corroborated the lipid nature of the gland secretion and showed in A. mellifera the poverty of the synthetic machinery in the worker gland cells in comparison with the queen, as expected by previous biochemical analyses. The pathway of the exogenous compounds of the secretion is intracellular, since substances can penetrate the cell folds and intercellular junctions, but their access to the, gland lumen is barred by the apical intercellular junctions.

Ultrastructural aspects of the mandibular gland of Melipona bicolor Lepeletier, 1836 (Hymenoptera : Apidae, Meliponini) in the castes

The mandibular gland in Melipona bicolor workers and queens was studied by scanning and transmission electron microscopy. There is no difference in the gland anatomy between the castes, but the transmission electron microscopy showed variation of the cellular ultrastructure according to the secretory phase of the gland in both castes. Smooth endoplasmic reticulum was abundant in the secretory cells of physogastric queens, indicating that these cells produce lipid secretion that is stored in granules with multi-lamellar bodies. Mitochondrial variations during the cell secretory cycle indicates their participation in the lipid synthesis. After secretion, release in the reservoir lumen through the collecting canals, the secretory cells contain many myelinic bodies, indicative of cellular regression. (C) 2004 Elsevier Ltd. All rights reserved.

MORPHOLOGICAL CASTE DIFFERENCES IN POLYBIA AEQUATORIALIS (HYMENOPTERA: VESPIDAE: POLISTINAE: EPIPONINI), A SOCIAL WASP of THE HIGHLANDS of COSTA RICA

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

História de vida de Laetacara aff. araguaiae Ottoni & Costa, 2009 (Perciformes, Cichlidae) em dois riachos no Noroeste do Estado de São Paulo, Brasil

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

Migration of the curimbatá Prochilodus lineatus (Valenciennes, 1836) (Pisces, Prochilodontidae) at the waterfall Cachoeira de Emas of the Mogi-Guaçu river - São Paulo, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Structural and ultrastructural analysis of embryonic development of Prochilodus lineatus (Valenciennes, 1836) (Characiforme ; Prochilodontidae)

This survey was performed to characterize the embryogenesis of Prochilodus lineatus. Seven stages of embryo development were identified - zygote, cleavage, blastula, gastrula, segmentation, larval and hatching - after a period of incubation of 22h (24 degrees C) or 14h (28 degrees C). The following cleavage pattern was identified: the first plane was vertical (2 blastomeres); the second was vertical and perpendicular to the first (4 blastorneres); the third was vertical and parallel to the first (4 x 2); the fourth cleavage was vertical and parallel to the second (4 x 4); the fifth was vertical and parallel to the first (4 x 8); and the sixth cleavage was horizontal (64 blastomeres). At the blastula stage (3.0-4.0 h (24 degrees C); 1.66-2.0h (28 degrees C) irregular spaces were detected and periblast structuring was initiated. At the gastrula stage (4.0-8.0 h (24 degrees C); 3.0-6.0 h (28 degrees C) the epiboly, convergence and cell movements, as well as the formation of embryonic layers, had begun. The segmentation stage (10.0-15.0h (24 degrees C); 7.0-10.0h (28 degrees C)) was characterized by a rudimentary formation of organs and systems (somites, optic vesicle and intestinal delimitation). The embryo at the larval stage (16.0-21.0 h (24 degrees C); 11.0-13.0 h (28 degrees C)) showed a free tail, more than 25 somites, an optic vesicle and a ready-to-hatch larval shape. The blastomeres at cleavage stage had disorganized nuclei indicating high mitotic activity. At gastrula, the blastomeres and the periblast had euchromatic nuclei and a large number of mitochondria and vesicles. The yolk was organized into globose sacs, which were dispersed into small pieces prior to absorption.

Factors associated with exclusive breastfeeding in children under four months old in Botucatu-Sp, Brazil

This study aimed to identify factors associated with exclusive breastfeeding (AME) and the reasons mothers presented to introduce complementary feeding in the first four months of life. A total of 380 mothers (92.2%) of children under four months old vaccinated in a Multi-vaccination Campaign were interviewed. To identify factors associated to AME, univariate and multiple logistic regressions analyses were performed. Thirty-eight percent of the children were on AME; 33.4% consumed cow milk; 29.2% tea; and 22.4% water. The mothers justified introduction of cow milk by factors related to quantity/quality of maternal milk and necessity of the child. The use of a pacifier (odds ratio=2.63; CI95%=1.7-4.06) and difficulty to breastfeed (odds ratio=1.57., CI95%=1.02-2.41) were associated with the absence of AME. The populational attributable risk percentage for the use of a pacifier was estimated at 46.8%. Thus, modifiable risk factors were associated with AME interruption.

Gli infortuni sul lavoro in Brasile. Studio nello Stato di San Paolo del Brasile, regione di Botucatu, utilizzando il metodo dell'albero delle cause

This paper discusses, within the prevaling Brazilian situation, the possibility of applying 'Causal Tree' (CT) method in investigating occupational accidents by safety personnel in the public health services and workers' unions. The method was developed during the seventies in France, for use by plant safety personnel. The authors used this method in Botucatu, State of Sao Paulo, Brazil, in order to investigate 40 serious occupational accidents that occurred in industrial plants during the second half of 1993, that had been registered by Social Security. In these cases, the predominance of situations in which the lack of safety measures were identified by inspection indicates that in most instances, the use of CT is unnecessary. However, the authors discuss its use by safety personnel from the public health services and workers' unions to investigate certain accidents to contribute to the knowledge base and help overcome the cultural based guilt which, in Brazil, has turned the victim into the person responsible for the accident.

Spermiophagy in the spermatheca of Melipona bicolor lepeletier, 1836 (Hymenoptera, Apidae, Meliponini)

The presence of spermatozoa in vesicles in the cytoplasm of the epithelial cells that constitute the spermathecal wall of Melipona bicolor queen (Meliponini) is discussed in relation to the organ structure. The epithelial wall is lined by an apparently continuous cuticle in the luminal surface that should be a non-transposable barrier to the luminal spermatozoa. However, some spermatozoa were seen crossing the cuticle through interruptions that was first interpreted as sectioning defects. Nevertheless, the sperm cells in well-structured cytoplasmic vesicles, bound by membranes and sometimes associated to multivesicular bodies, as well as cytoplasmic structures representative of intracellular digestion and the occurrence of the phenomenon in two of the three spermathecae studied, suggest a real spermiophagic hole in the spermathecal epithelial cells.