Desenvolvimento morfolôgico dos ovários em embriões e fetos bovinos da raça Nelore

The morphologic events related to the prenatal development of the ovaries in 81 Nelore breed embryos and fetuses gathered in a local slaughterhouse, with age range from 26 to 240 days following fecundation were studied. The age of fetuses was estimated from measures taken in the cranium-caudal direction. The sex was identified from macroscopic observations and using Polymerase Chain Reaction (PCR) technique. For histology the gonads were fixed in Bourn's fluid for 24 hours and 5 μm thick section's were stained with hematoxylin-eosin. Formation of gonadal ridge and presence of germinal cells were found within it at 29 and 34 days, respectively. Oogonia and primordial follicles, unlike the growing follicles, exhibited significant differences in diameter in the various periods studied. Positive correlation (P<0.05) was found between the diameter of oogonia and their nucleus as well as between primordial and growing follicles with their oocytes and respective nuclei. The gonad was fully formed at 40 days. Primordial follicles, in the growing stage, and antral follicles first appeared, approximately at 95, 140, and 180 days, respectively. Despite the onset and duration of oogenesis being similar to that of taurine breeds, folliculogenesis initiates at an early stages in the Nelore breed.

Diagnosis of 5α-reductase type 2 deficiency: Contribution of anti-Müllerian hormone evaluation

Aim: To evaluate anti-Müllerian hormone (AMH) levels in patients with clinical and molecular diagnosis of 5α-reductase 2 deficiency. Patients and methods: Data from 14 patients whose age ranged from 21 days to 29 years were analyzed according to age and pubertal stage. Sexual ambiguity was rated as Prader III in 11 patients. LH, FSH, testosterone (T), dihydrotestosterone (DHT) and AMH serum levels were measured in all but two patients, who had been previously submitted to gonadectomy; T and DHT were also measured in 20 age-matched controls. Results: Gonadotropin levels were normal in all but one patient who retained gonads (six of whom had reached puberty) and T/DHT ratio was elevated in all patients when compared to controls. All prepubertal patients had AMH levels < -1 SD for age, while most pubertal patients had AMH levels compatible with pubertal stage. Conclusions: Prepubertal patients with 5α-reductase 2 deficiency have AMH values in the lower part of the normal range. These data indicate that T does not need to be converted to DHT to inhibit AMH secretion by Sertoli cells. © Freund Publishing House Ltd., London.

Aspectos da reprodução no gênero Characidium Reinhardt, 1867 (Crenuchidae, Characidiinae), na microbacia do Ribeirão Grande, serra da Mantiqueira, sudeste do Brasil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Gonad development in females of fiddler crab Uca rapax (Crustacea, Brachyura, Ocypodidae) using macro and microscopic techniques

The morphology of the ovaries in Uca rapax (Smith, 1870) was described based on macroscopic and microscopic analysis. Females were collected in Itamambuca mangrove, Ubatuba, state of São Paulo, Brazil. In the laboratory, 18 females had their ovaries removed and prepared for histology. Each gonad developmental stage was previously determined based on external and macroscopic morphology and afterwards each stage was microscopically described. The ovaries of U. rapax showed a pronounced macroscopic differentiation in size and coloration with the maturation of the gonad, with six ovarian developmental stages: immature, rudimentary, developing, developed, advanced and spent. During the vitellogenesis, the amount of oocytes in secondary stage increases in the ovary, resulting in a change in coloration of the gonad. Oogonias, primary oocytes, secondary oocytes and follicular cells were histologically described and measured. In female's ovaries of U. rapax the modifications observed in the oocytes during the process of gonad maturation are similar to descriptions of gonads of other females of brachyuran crustaceans. The similarities are specially found in the morphological changes in the reproductive cells, and also in the presence and arrange of follicle cells during the process of ovary maturation. When external morphological characteristics of the gonads were compared to histological descriptions, it was possible to observe modifications that characterize the process in different developmental stages throughout the ovarian cycle and, consequently, the macroscopic classification of gonad stages agree with the modifications of the reproductive cells.

The reproductive biology and cycle of Rioraja agassizi (Müller and Henle, 1841) (Chondrichthyes: Rajidae) in southeastern Brazil, SW Atlantic Ocean

Specimens of Rioraja agassizi were collected at Guarujá pier, São Paulo State, Brazil, from March 2005 to March 2006. A total of 275 males were captured. Based on the analysis of the clasper length, gonad weight, clasper gland length, alar thorns and lobule diameter, the size-at-maturity was estimated to be 32 cm. There were 1049 females in the record. Size-at-maturity was calculated in 40 cm. Sexual resting females were observed. Gonadosomatic and hepatosomatic indexes varied significantly in both sexes throughout the year. However, ovulation, egg-laying and presence of sperm in the seminal vesicle were observed all year round. For this reason, an annual cycle with at least one peak in the sexual activity is proposed for this species.

Morfogênese do testículo de embriões e fetos de vacas da raça nelore (Bos taurus indicus)

The aim of this study was to accompany the process of testicular development from the non-differentiable phase to its complete formation. Embryos and fetuses of Nelore breed cows (Bos Taurus indicus) were obtained in slaughterhouses near the Uberlandia city, Minas Gerais. The gonads and the embryos were fixed in Bouin's fixative and afterwards processed for conventional optical microscopy. The gonadal was observed firstly in a 1.0 cm long embryo. In 2.5 cm long embryos the presence of the albuginea allows the sex identification. The mean thickness of the albuginea ranged from 29.08 to 558.45 mm. Gradually increase of vascularization of the albuginea and parenchyma is observed. The mediastinum is located centrally. There was a decrease in the space occupied by the testicular cords, from 63.71 to 41.99% of the total testes volume. Its diameter ranged from 31.68 to 48.80 mm. The diameter of germinal cells (and their nuclei) was from 12.27 (6.65) to 16.95 914.21) mm. The quantity of germinal cells by cross section of cord decreased from a maximum of 2.80 to 0.76. The total number of germinal cells was from 16 at the beginning of colonization of the gonad to 18.32 x 106 at the end of the study. The number of Sertoli's cells by cross section of cord ranged from 10.00 to 16.25. The results obtained show that the origin and formation of testes in embryos and fetuses from Nelore breed cows (Bos taurus indicus) does occur in a very similar way to what is described for Bos taurus taurus.

Sexual development and reproductive cycle of the Eyespot skate Atlantoraja cyclophora (Regan, 1903) (Chondrichthyes: Rajidae: Arhynchobatinae), in southeastern Brazil

Specimens o/Atlantoraja cyclophora were collected monthly from commercial fishing landings at Guarujá, São Paulo State, Brazil, from March 2005 to April 2006 at depths between 10 and 146 m. Males ranged from 13.3 to 58.5 cm TL (n = 396). Both the smallest mature male and the largest immature male were 47.0 cm long. Males' size-at-50% maturity was calculated to be 46.3 cm. Females ranged from 11.5 to 68.0 cm (n = 401). The smallest mature and the largest immature female were 51.6 and 53.0 cm long respectively. For the females, size-at-50% maturity was calculated to be 53.2 cm. In the males the hepatosomatic andgonadosomatic indices varied between 0.48 (August) and 3.54 (November) and between 0.15 (November) and 1.45 (June) respectively, with no significant variation for the fourteen-month period. In the females the hepatosomatic and gonadosomatic indices varied from 1.55 and 6.30 3.54 (both for April 2006) and from 0.08 (December) to 4.41 (October) respectively, with no significant difference among months. Egg-bearing females were found in all months with proportions varying from 0.03 (March) to 0.67 (April). Both males and females undergo an annual cycle, with slight seasonal variations in reproductive activity and a peak in the proportion of egg bearing females between April and July.

Cellular localization of androgen synthesis in equine granulosa-theca cell tumors: Immunohistochemical expression of 17α-hydroxylase/17,20-lyase cytochrome P450

Elevated blood testosterone concentrations, often accompanied by male-typical behaviors, is a common signalment of mares with granulosa-theca cell tumors (GCTCs), but no definitive information exists regarding the cellular differentiation of tumors associated with androgen secretion. This study was conducted to localize and thereby define the cellular expression of 17α-hydroxylase/17,20-lyase cytochrome P450 (P450c17), the enzyme most directly responsible for androgen synthesis, in 30 GTCTs and control tissues (gonads and adrenal glands) using immuno-histochemistry (IHC). Immuno-reactivity for P450c17 was evident in approximately half of 30 specimens examined, was most consistent in the interstitial cells surrounding existing or developing cysts, and was less intense in cells within cysts in the smaller proportion of specimens where this was observed. In control tissues, the expression of P450c17 was localized primarily in theca interna of normal ovarian follicles, in theca-lutein cells of some corpora lutea, but not in granulosa-lutein cells. Testicular interstitial cells and islands of adreno-cortical cells located in the adrenal medulla of the adrenal cortex further established the specificity of the antisera used. These data provided the first substantive evidence that polyhedral cells identified previously in GTCTs by histopathology have the potential to synthesize and secrete androgens, similar to theca interna and theca lutein cells in normal equine ovaries. © 2010 Elsevier Inc.

Reproduction, food dynamics and exploitation level of Oreochromis niloticus (Perciformes: Cichlidae) from artisanal fisheries in Barra Bonita Reservoir, Brazil

Nile tilapia (Oreochromis niloticus), which is exotic to South America, is the most common species caught in artisanal fisheries at the Barra Bonita Reservoir, Southeastern Brazil. This species is of great socioeconomic importance for the region and keeps active a population of about 500 fishers. In the present study we assess reproduction, food dynamics and level of exploitation of O. niloticus, caught by artisanal fisheries in the Barra Bonita Reservoir. Specimens were collected monthly, from July 2004-June 2005, and a total of 1 715 specimens were analyzed. Each specimen was examined to obtain biological and biometric data: standard length (cm), total weight (g), reproductive data (sex and stage of maturation), and stomach contents (empty, partly full, and full). We also estimated the sex ratio (by macroscopic observation of gonads), reproductive period (by ovarian development and seasonal average of gonadosomatic index in females), and feeding habits (by stomach contents). The possible relationship between abiotic factors and the reproductive period was statistically verified using Spearman's Rank Correlation. The FiSAT (ELEFAN I) package was used to assess growth parameters, mortality rates and to infer exploitation rate from standard length frequencies. The O. niloticus population had a sex ratio of 1.3:1 (M:F). Results indicated that ripe females were captured throughout the year, with a higher frequency during the winter-2004 (with a frequency of 59%, at a mean temperature of 20.5°C), and in spring-2004 (with a frequency of 60.5% at a mean temperature of 21.18°C). The GSI mean values obtained by season were: winter-2004: 1.71; spring-2004: 1.72; summer-2005: 0.80, and autumn-2005: 1.19. The Spearman correlation indicated positive values with respect to pH, dissolved oxygen, electric conductivity, transparency and chlorophyll a, and negative values with respect to temperature, accumulated rainfall and altimetric benchmark. The main food items were phytoplankton and periphytic algae, observed in 99.6% of the analyzed stomachs. The estimated growth and mortality parameters were: L∞=33.60cm, k=0.63/year, longevity= 4.76years, Z=2.81/ year, M=1.20/year and F=1.61/year. The weight-length relationship was Ln Wt=-2.8532+2.8835 Ln Lp. The estimated yield per recruit values were as follows: E=0.570, Emax=0.776, E0.1=0.604 and E0.5=0.349. These results indicate that a well established population of O. niloticus is present at Barra Bonita Reservoir; with an active reproduction throughout the year, more intense during winter and spring, and that O. niloticus is a phytoplanktophagus species. There were no indications that this species is being overfished, we therefore recommend that, due to its exotic condition, no restrictions need to be taken on its fishing activities.

Gametogenesis and reproductive cycle of Melanorivulus aff. punctatus (Boulenger, 1895) (Cyprinodontiformes, Rivulidae) in Chapada dos Guimarães, Mato Grosso, Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Female germ cell renewal during the annual reproductive cycle in Ostariophysians fish

The objective was to characterize female germ cell renewal during the annual reproductive cycle in two species of ostariophysian fish with distinct reproductive strategies: a siluriform, Pimelodus maculatus, in which oocyte development is group synchronous and the annual reproductive period is short; and a characiform, Serrasalmus maculatus, with asynchronous oocyte development and a prolonged reproductive period. These reproductive strategies result in fish determinate and indeterminate fecundity, respectively. Annual reproductive phases were determined by biometric and histologic analysis of gonads and interpreted according to new proposals for phase classification and stages of oocyte development (with special attention to germinal epithelium activity). Histologically, there were two types of oogonia in the germinal epithelium: single oogonia and those in mitotic proliferation. Oogonial proliferation and their entry into meiosis resulted in formation of cell nests (clusters of cells in the ovarian lamellae). Morphometric analysis was used to estimate germ cell renewal. Based on numbers of single oogonia in the lamellar epithelium, and nests with proliferating oogonia or early prophase oocytes throughout the annual reproductive cycle, oogonial proliferation and entrance into meiosis were more intense during the regenerating phase and developing phase, but decreased sharply (P < 0.05) during the spawning-capable phase. Oogonial proliferation gradually recovered during the regressing phase. We concluded that, independent of species or features of the reproductive cycle, germ cell renewal occurred during the regenerating phase, ensuring availability of eggs for the spawning event. © 2013 Elsevier Inc.

Population dynamics of the freshwater crab Dilocarcinus pagei in tropical waters in southeastern Brazil

In the northwestern region of the state of São Paulo, Brazil, the freshwater crab Dilocarcinus pagei Stimpson, 1861 is usually sold as bait for several fish species for sport fishing. Thus, knowledge of the reproductive biology of this species is essential to provide information for maintaining natural stocks and implementing future farming initiatives. The objective of this study was to investigate the population dynamics of the red freshwater crab D. pagei, focusing on reproductive and juvenile recruitment periods, sex ratio, and individual growth. Sampling was performed monthly from October 2005 to September 2007 in the Marimbondo-Fumas Hydroelectric Plant reservoir, Rio Grande, municipality of Icém in northwestern São Paulo state. The individuals were sexed, carapace widths were measured, and the gonads were examined macroscopically. In total, 1002 individuals were analyzed, consisting of 568 males and 434 females, of which 4 were ovigerous and 35 carried early juveniles in the incubatory chamber. The largest numbers of reproductive individuals of both sexes were recorded during the winter and spring months before spawning, which occurred in the spring. Intense juvenile recruitment and the highest rainfall levels were observed during the Southern Hemisphere summer. The results indicate that spawning can be considered seasonal, and the simultaneous emergence of juveniles with high rainfall levels is probably the result of the higher availability of food and potential sites for protection against predators. The skewed sex ratio (♂ 1:0.76 ♀) may be associated with growth rates, mortality, food restriction, behavioral differences, migration, and differential habitat use between the sexes. The similar maximum size (CW∞) and growth constant (k) for males (k = 1.78 y-1, CW = 61.43 mm) and females (k = 1.67 y-1, CW = 60.66 mm) may be associated with different energy investment strategies between the sexes. Additionally, a closed season is proposed for the species. © The Crustacean Society, 2013. Published by Brill NV, Leiden.

Population dynamics of the seabob shrimp Xiphopenaeus kroyeri (Dendrobranchiata, Penaeidae) in south-eastern Brazil

The population dynamics, including the sex ratio, reproductive period, individual growth and longevity, and population structure of the shrimp Xiphopenaeus kroyeri, are described. The shrimps were collected monthly from July 2005 to June 2007 at four sites in Ubatuba Bay, Brazil. The salinity, temperature, depth, organic matter content and sediment grain size were measured at each site. Specimens were quantified and sexed, their carapace length measured and their gonads were classified according to the stage of maturation. A von Bertalanffy model was used to estimate the individual growth, and longevity was calculated using its inverted formula. In total, 6 470 individuals were examined and the number of males and females differed significantly in favour of females. During some periods, the segregation could be related to the reproductive period. Reproductive females were captured throughout the sampled period, and there were biannual peaks that were followed by peaks of juveniles. Carapace growth was less for males than for females, which is likely related to oocyte production and the higher fecundity of the females. The higher probability of female capture can be related to their longer foraging to sustain the enhanced food requirement during the ovarian maturation period. © 2013 Copyright NISC (Pty) Ltd.

Cyp17a1 and cyp19a1 in the zebrafish testis are differentially affected by oestradiol

Oestrogens can affect expression of genes encoding steroidogenic enzymes in fish gonads. However, little information is available on their effects at the protein level. In this context, we first analysed the expression of key steroidogenic enzyme genes and proteins in zebrafish testis, paying attention also to other cell types than Leydig cells. Gene expression was analysed by quantitative PCR on fluorescence-activated cell-sorting fractions coupled or not to differential plating, while protein synthesis was studied by immunohistochemistry using specific antibodies against zebrafish Cyp17a1, Cyp19a1a and Cyp19a1b. Furthermore, we have evaluated the effect of oestrogen treatment (17β-oestradiol (E2), 10 nM) on the localization of these enzymes after 7 and 14 days of in vivo exposure in order to study how oestrogen-mediated modulation of their expression is linked to oestrogen effects on spermatogenesis. The major outcomes of this study are that Leydig cells express Cyp17a1 and Cyp19a1a, while testicular germ cells express Cyp17a1 and both, Cyp19a1a and Cyp19a1b. As regards Cyp17a1, both protein and mRNA seem to be quantitatively dominating in Leydig cells. Moreover, E2 exposure specifically affects only Leydig cell Cyp17a1 synthesis, preceding the disruption of spermatogenesis. The oestrogen-induced suppression of the androgen production capacity in Leydig cells is a major event in altering spermatogenesis, while germ cell steroidogenesis may have to be fuelled by precursors from Leydig cells. Further studies are needed to elucidate the functionality of steroidogenic enzymes in germ cells and their potential role in testicular physiology. © 2013 Society for Endocrinology.

Reproductive biology of two macrourid fish, Nezumia aequalis and Coelorinchus mediterraneus, inhabiting the NW Mediterranean continental margin (400-2000m)

Nezumia aequalis and Coelorinchus mediterraneus are abundant species on the upper and lower continental slopes, respectively, in the Mediterranean Sea. A study on the reproductive strategy of the two species was conducted on the Catalan margin (NW Mediterranean). The reproductive cycle of both species was investigated using visual analyses of gonads and histological screening. The shallower species, N. aequalis, showed continuous reproduction with a peak of spawning females in winter months. In contrast, the deeper-living species, C. mediterraneus, showed semi-continuous reproduction with a regression period during the spring. Juveniles of N. aequalis were present in all seasons, but most abundant in the spring. Only two juveniles of C. mediterraneus were found. Both species had asynchronous oocyte development. The average fecundity of N. aequalis was 10,630 oocytes per individual, lower than known for the same species in the Atlantic Ocean. The fecundity of C. mediterraneus was measured for the first time in this study, with an average of 7693 oocytes per individual. Males of both species appear to have semi-cystic spermatogenesis. © 2013 Elsevier Ltd.