Chronic estrone exposure affects spermatogenesis and sperm quality in zebrafish (Danio rerio).

Estrone (E1) is a common environmental contaminant found in rivers and streams due to the farming of animals, such as swine and cattle. Our study evaluated the effects of chronic E1 exposure at environmentally relevant concentrations on spermatogenesis and the semen quality of zebrafish (Danio rerio). We exposed the fish to E1 at concentrations of 20, 200, and 2000 ng/L diluted in 0.001% ethanol (v/v) for 49 days. There were two control groups: one was exposed to water only and the other to ethanol at the same concentration used in the E1 groups. Following exposure, we analyzed the proportion of testicular cell types and other components (%), rate of cell proliferation and death, and sex steroid concentrations. Furthermore, we analyzed the expression of insulin-like growth factor 1 (IGF1), IGF2, IGF1 receptor (IGF1R), and inducible nitric oxide synthase and assessed the semen quality. E1 exposure increased spermatogonia, spermatids, Sertoli cells, Leydig cells, and the proportion of inflammatory infiltrate but decreased the spermatozoa amount. These changes were reflected by reductions in the gonadosomatic index and levels of 11-ketotestosterone in the testes. On the other hand, E1 exposure increased testicular estradiol, IGF1R expression, and nitric oxide production. After an evaluation using a computer-assisted sperm analysis (CASA) system, we observed reduced progressive motility, curvilinear velocity, and beat cross frequency of 20 and 2000 ng/L E1 groups. Our findings support that E1 causes deleterious effects on the testicular function and semen quality of D. rerio even at environmental concentrations. Thus, E1 concentrations should be monitored in surface waters for the purposes of fish conservation.

Effects of temperature manipulation on gamete development and reproductive activity in the farmed catfish Lophiosilurus alexandri.

Temperature is a critical factor for fish management, especially during breeding season. This study evaluated gametogenesis and gonadal maturation in the catfish Lophiosilurus alexandri under farming conditions in two temperatures. The fish were divided in the following experimental groups: (S0) control group, before starting the experiment, (S1) 30 days at 23 °C, (S2) 70 days at 23 °C, (S3) 30 days at 27 °C, and (S4) 70 days at 27 °C. Gonad, liver, and blood samples were obtained to analyze reproductive and health parameters. In females, increased gonadosomatic index (GSI) was associated with a decreased proportion of perinucleolar follicles and increased diameter of vitellogenic follicles in S4. Postovulatory follicles were found only in fish kept at 27 °C, whereas atretic follicles were more frequent at 23 °C. In S4, females had higher hepatic levels of vitellogenin and lower levels of zona radiata proteins. In males, a lower proportion of spermatogonia was found at 27 °C (S4), whereas GSI and proportions of spermatocytes, spermatids, and spermatozoa were similar among the groups. Levels of cortisol were higher in females of group S3. Plasma 17ß-estradiol, 11-ketotestosterone and 17α-hydroxyprogesterone were significantly higher in S4. Hematocrit, erythrocytes, and glucose were higher at 27 °C. Together, the present study indicate that exposure of L. alexandri to 27 °C provides suitable conditions for germ cell development and gonadal maturation in captivity, promotes increase of sex steroids, follicular growth, and oocyte maturation, with onset of ovulation occurring at 30 days. At 23 °C, spermatogonia proliferation is favored in males, but it cannot be recommended for stimulating female maturation of L. alexandri in fish farming.

Adverse effects of herbicides in freshwater Neotropical fish: A review.

Although herbicides have been developed to act on the physiological processes of plants, they are responsible for causing deleterious effects on animals. These chemical compounds are widely used throughout the world, but especially in countries that export agricultural products such as Central and South America, their use has increased in recent years. Aquatic environments are natural reservoirs of herbicides, which after being applied on crops, run off through the soil reaching rivers, lakes, and oceans. Fish are among the many organisms affected by the contamination of aquatic environments caused by herbicides. These animals play an important ecological role and are a major source of food for humans. However, few studies address the effects of herbicides on fish in this region. Thus, in the present review we discuss the morphophysiological and molecular consequences of herbicide exposure in Neotropical fish systems as well as how the environmental and land use characteristics in this region can influence the toxicity of these pollutants. A toxicity pathway framework was developed summarizing the mechanisms by which herbicides act and endpoints that need to be further investigated.

The metabolite p-cresol impairs dendritic development, synaptogenesis, and synapse function in hippocampal neurons: Implications for autism spectrum disorder.

Autism spectrum disorder (ASD) is a heterogeneous neurodevelopment disorder resulting from different etiological factors, both genetic and/or environmental. These factors can lead to abnormal neuronal development on dendrite and synaptic function at the central nervous system. Recent studies have shown that a subset of ASD patients display increased circulation levels of the tyrosine metabolite, p-cresol, related to chronic intestinal disorders because of dysbiosis of the intestinal microbiota. In particular, abnormal presence of intestinal Clostridium sp. has been linked to high levels of p-cresol in ASD children younger than 8 years. However, the role of p-cresol during development of the central nervous system is unknown. Here, we evaluated in vitro the effect of p-cresol on neurite outgrowth in N2a and PC12 cell lines and dendritic morphology, synaptic density, neuronal activity, and calcium responses in primary rat hippocampal neurons. p-cresol inhibits neural differentiation and neurites outgrowth in N2a and PC12 neuronal cell lines. In hippocampal neuronal cultures, Sholl's analysis shows a decrease in the dendritic arborization of neurons treated with p-cresol. Synaptic density analyzed with the synaptic markers Piccolo and Shank2 is diminished in hippocampal neurons treated with p-cresol. Electrically evoked intracellular calcium rise was drastically, but reversely, blocked by p-cresol, whereas that spontaneous neuronal activity was severely affected by early addition of the metabolite. These findings show that p-cresol alters dendrite development, synaptogenesis, and synapse function of neurons in culture, therefore, neuronal alterations occurring in ASD children may be related to this metabolite and dysbiosis of the intestinal microbiota.

Biomarker responses induced by bisphenol A on spermatogenesis in a Neotropical teleost fish are temperature-dependent.

Bisphenol A (BPA) is an organic synthetic compound used in the plastic industry with endocrine disrupting activity. Although it is frequently found in surface waters, few studies have investigated its impact on fish gametogenesis, particularly when associated with natural stressors. In this regard, the present study evaluated BPA toxicity on spermatogenesis in the lambari Astyanax bimaculatus under controlled conditions and its interactive effects with water temperature. Adult specimens were exposed in duplicate to 40 µg/L and 400 µg/L BPA at 23 °C and 28 °C for 21 days; the control group did not receive BPA. Testicular samples were collected and analyzed using different cellular and molecular techniques. The results showed a significant reduction in the gonadosomatic index in the BPA-treated groups at both temperatures. A decrease in the testicular levels of 11-ketotestosterone was observed in the 400 µg/L BPA group at 23 °C, 17ß-estradiol increased significantly in the treated groups at 28 °C, and vitellogenin showed no difference between the treatments. The morphometric analysis of spermatogenesis revealed a significant increase in the proportion of spermatogonia, spermatocytes, and Sertoli cells in the treated groups, with a higher proportion at 23 °C than at 28 °C. Otherwise, the proportion of spermatozoa was significantly lower in the BPA-treated groups, with a greater reduction at 23 °C. In addition, BPA also stimulated spermatogonial proliferation in the treated groups, but apoptosis was significantly increased in spermatids at 23 °C. Testis-ova, cell degeneration, and chromatin alterations in spermatids and Sertoli cells were observed in the germinal epithelium of the BPA-treated groups. The integrated biomarker response (IBR) index revealed that the analyzed endpoints are suitable for assessing estrogenic contamination. Taken together, our results indicate that the interactive effects of BPA and temperature contribute to the impairment of spermatogenesis in A. bimaculatus with more severe effects observed on sperm production at 23 °C than at 28 °C.

Evaluation of the oestrogenic potential of oestrone and bisphenol-A on the reproduction of Astyanax bimaculatus males after subacute exposure.

In the last decades, oestrogenic compounds have often been reported in environmentally relevant concentrations in aquatic environments around the world. Most laboratory studies of oestrogens try to understand the effects of a single contaminant, but in natural environments, the effects may be quite different due to interactions with other compounds. The present study aimed to compare the action of oestrone (E1) and bisphenol-A (BPA), acting singularly and in combination, on the spermatogenesis of Astyanax bimaculatus. After exposure to 100 ng/L of E1, BPA and a mixture of the two for 15 days, our results showed that E1 and the E1 + BPA mixture significantly altered the number of spermatogenic cells. BPA presented high cytotoxicity when compared to other treatments. Analysis of the two oestrogenic compounds suggests that the E1 + BPA mixture has no additive or synergistic effects. Together, the results of the present study indicate that endocrine-disrupting chemicals (EDCs) analysed alone may behave differently than when administered with other substances.

Insulin-like growth factors 1 and 2 are associated with testicular germ cell proliferation and apoptosis during fish reproduction.

To support sperm production, fish testes undergo intense tissue remodelling, with endocrine, paracrine and autocrine signals regulating gonad physiology. The aim of this study was to investigate the testicular expression of insulin-like growth factor (Igf) 1 and Igf2 during spermatogenesis, and their relationship with cell proliferation and apoptosis throughout the reproductive cycle. The study was performed in male Hypostomus garmani, a catfish living in headwater rivers of the São Francisco River basin, Brazil. Spermatogenesis was analysed using histology, morphometry, immunohistochemistry and terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL) analysis at different maturity stages. The results showed the proliferation of spermatogonia throughout the reproductive cycle, with a higher rate during the ripe stage. Germ and Sertoli cells expressed Igf1 at all stages of testicular maturity, Igf2 was predominant at the ripe stage and both Igf1 and Igf2 occurred at the spent stage. Caspase-3 and TUNEL analysis revealed a higher rate of apoptosis at the spent stage associated with reduced expression of Igf1 and Igf2. Sertoli cell proliferation was associated with spermatogonia and spermatocyte cysts at different stages of the reproductive cycle. Together, the data support a proliferative role for Igf1 and Igf2 in regulating testicular apoptosis in H. garmani, with cyclical variation in their expression during gonad maturation.

Effects of metal contamination on liver in two fish species from a highly impacted neotropical river: A case study of the Fundão dam, Brazil.

Environmental disasters such as the rupturing of mine tailings dams are a major concern worldwide. In the present study, we assess the effects of the release of mine waste due to the rupture of the Fundão dam on two native fish species (Hoplias intermedius and Hypostomus affinis) from the Doce River basin. Two sampling sites were chosen: S1, a reference site, and S2, contaminated by mining waste. Water and sediment were collected to evaluate metals concentration. Adult fish were caught to analyse biological parameters, hepatic histopathology, and biomarkers of metal contamination. Compared to site S1, the concentration of manganese was statistically higher in water while lead, nickel, and arsenic were statistically higher in the sediment from site S2, and iron had no significant difference between sites. At site S1, fish of both species presented hepatic tissue with normal architecture. At site S2, hepatic alterations, such as cytoplasmic vacuolization and necrosis were frequently found in both species. Regarding the histopathological index, higher values were found in both species from site S2. The positive antibody reactions for cytochrome P450 1A (CYP1A) and metallothionein (MT) were statistically greater in site S2 for both species. The oxidative stress biomarkers, superoxide dismutase (SOD) and catalase (CAT) were statistically higher in H. intermedius from site S2, but only CAT was statistically greater in H. affinis at site S2. These results demonstrate that the release of mineral residues from the rupture of the Samarco mine dam is provoking hepatic damage in the fish from the Doce River besides inducing the expression of proteins and enzymes related to metal contamination.

Effects of starvation and refeeding cycles on spermatogenesis and sex steroids in the Nile tilapia Oreochromis niloticus.

Food restriction is part of the life cycle of many fish species; however, nutritional deficiency may negatively influence gametogenesis and gonadal maturation. The aim of this study was to evaluate the effects of food restriction on the spermatogenesis of Nile tilapia. For this, adult males were submitted to starvation and refeeding cycles (alternating periods of starvation and feeding) for 7, 14, 21, and 28 days. After 7 days of starvation, glycaemic and lipid levels were significantly reduced, followed by reduction of plasma testosterone (T) and 11-ketotestosterone (11-KT). In addition, reduced proliferation of spermatogonia and increased apoptosis of spermatocytes, spermatids, and spermatozoa was observed in starvation groups. In the refeeding groups, the sex steroids and the proportion of germ cells had no significant alterations compared to the control group, except for spermatozoa. In this sense, the present study suggests that starvation after 7 days progressively reduces T and 11-TK, resulting in damage to the production of spermatogenic cells, while refeeding may delay spermatogenesis but does not lead to testicular impairment.

Influence of salinity on spermatogenesis in adult Nile tilapia (Oreochromis niloticus) testis.

Continental waters salinisation is a global threat that has grown because of climate change and human activities, but little is known about how and what biological tracts are affected. The aim of this study was to investigate the influence of different water salinities on the expression of HSP70, PCNA and caspase-3 during spermatogenesis of Nile tilapia. Adult males were submitted to four salinity treatments: (S0) fresh water, (S7) 7 g L-1, (S14) 14 g L-1, and (S21) 21 g L-1 for 1, 4, and 9 days. All specimens were in spermatogenic activity and the highest values of the gonadosomatic index (GSI) occurred in the S0 and S7. In the morphometric analysis, spermatocytes were the most frequent germ cell detected in all treatments (>50%) and spermatids achieved about 20% of the testicular proportion, with few variations among treatments. Spermatozoa were significantly reduced only in S14 compared to S7. Leydig cells were significantly increased in S14 when compared to S7 but plasma concentrations of 11-KT showed no significant difference among treatments. ELISA assay showed higher testicular expression of HSP70 at 1 day in all groups, followed by a significant decrease at days 4 and 9 in S14 and S21. The expression of PCNA was significantly lower while the activity of caspase-3 was higher in S14 and S21 when compared to S0 and S7. These results indicate that higher salinities in S14 and S21 interfere with the relationship between testicular HSP70, PCNA, and caspase-3, but with few effects over spermatogenesis dynamics of Nile tilapia.

Environmental exposure to oestrogenic endocrine disruptors mixtures reflecting on gonadal sex steroids and gametogenesis of the neotropical fishAstyanax rivularis.

Discharge of municipal wastewater promotes the entry of diverse oestrogenic compounds into the water bodies. This complex mixture of substances interferes in the steroidogenic pathway, being able to promote severe reproductive impairment in freshwater fish populations. The purpose of the present study was to evaluate the effects of oestrogenic endocrine disruptors (EDCs) mixture on gonadal sex steroids (testosterone, T; 11-ketotestosterone, 11-KT; 17ß-oestradiol, E2; 17-hydroxyprogesterone, 17-OHP) in the peak of the reproductive season of Astyanax rivularis, correlating the results obtained with the proportion of germ cells and gonadal histopathology. Three sampling sites were chosen to conduct the study, one reference site (S1), without contamination by municipal wastewater and two sites (S2 and S3) receiving discharge of municipal wastewater. Males of A. rivularis presented higher concentrations of E2, lower androgens (T and 11-KT) in gonads when compared to males from site S1. Concentrations of 17-OHP did not present significant difference among sites. In sites S2 and S3, the proportion of early spermatocytes, spermatids and Leydig cells increased while spermatozoa decreased compared to fish from S1. The following gonadal histopathologies were detected in the male fishes: intersex gonads (28% in S3) and testicular degeneration with germinal epithelium exhibiting agglutinated germ cells masses and empty cysts (57% in S2 and 71% in S3). In females, concentrations of T, E2 and 17-OHP did not present significant difference among the sites, however higher 11-KT concentrations were detected in females from sites S2 and S3. A lower proportion of perinucleolar follicles and a higher incidence of vitellogenic follicles, besides, aged oocytes and the presence of eosinophilic proteinaceous fluid in the interstitial compartment were also found in females from impacted sites. These results indicate that the urbanization and consequent release of municipal wastewater containing oestrogenic compounds in the headwater creeks are altering the levels of sex hormones and gametogenesis of A. rivularis. Further studies should be performed to determine whether oestrogenic endocrine disrupters are disrupting the reproduction of A. rivularis.

Expression patterns and immunolocalisation of IGF-I and IGF-II in male and female gonads of the Neotropical characid fish Astyanax fasciatus.

The insulin-like growth factor (IGF) system plays important roles in fish reproduction, but the expression pattern and cellular location of IGF-I and IGF-II during gonadal maturation are uncertain. The present study reports a stage-specific assessment of gonadal expression levels and immunolocalisation of IGF-I and IGF-II in Astyanax fasciatus, a characid fish from South America. Adult fish in different maturity stages were caught in the Furnas Reservoir, Grande River, Brazil. Gonad samples were processed for histology, immunohistochemistry, and ELISA for IGF-I and IGF-II. Ovarian levels of IGF-I were low during ripening and ripe stages, higher in totally spent, and then decreased in resting. Levels of IGF-II increased during ovarian maturation, reaching significantly higher values at stage totally spent. In males, IGF-I levels followed gonadal maturation, with higher values in ripening and ripe stages, whereas IGF-II levels showed higher values in stage ripening and partially spent. A positive correlation was found between IGF-I and gonadosomatic index (GSI) for males (r = 0.59), while females showed a negative correlation (r = - 0.43), but IGF-II showed no correlation to GSI. IGF-I was expressed mainly in oogonia nests whereas IGF-II stained the follicular cells in the perinucleolar follicles, cortical vesicles in the previtellogenic follicles, and oogonia nests. In males, IGF-I was evident in spermatogonia and spermatocytes while IGF-II stained Sertoli cells surrounding spermatids cysts and spermatogonia in late stages. Together, these findings support a hypothesis that the balance between IGF-I and IGF-II levels is important in the regulation of gonad maturation in Astyanax fasciatus.

Stage-specific testicular protein levels of the oestrogen receptors (ERα and ERß) and Cyp19 and association with oestrogenic contamination in the lambari Astyanax rivularis (Pisces: Characidae).

Oestrogens participate in various biological processes such as oogenesis, vitellogenesis and testicular development, but studies regarding the distribution and protein levels of oestrogen receptors (ERα and ERß) and aromatase (Cyp19) in testis are rarely investigated in fish species. The aim of the present study was to analyse the expression pattern of ERα, ERß and Cyp19 in testis of Astyanax rivularis and, in addition, to verify if oestrogenic contamination interferes in the expression levels of these proteins. Quarterly, field samplings were carried out during a reproductive cycle in a stream of the Upper Velhas River with a good conservation status (site S1). In the gonadal maturation peak (June), when ripe stage was most abundant, fish collection was made in three streams: S1, reference site, and S2 and S3, sites contaminated by untreated sewage. The results of immunohistochemistry demonstrated labelling of Cyp19 in Leydig cells and acidophilic granulocytes, but spermatogonia, Sertoli cells, spermatids and spermatozoa were also labelled. ERα was more widely distributed than ERß being found in all developmental germ cell phases. On the other hand, ERß was found only in spermatogonia and spermatocytes. During testicular maturation, ELISA levels for Cyp19, ERα and ERß followed the gonadosomatic index (GSI) with significant higher values in the ripe stage. Regarding to endocrine disruption, the males exposed to domestic sewage presented significant higher expression of Cyp19 and ERα when compared to the non-exposed fish. Together, our results demonstrate expression patterns of Cyp19, ERα and ERß in the testis of A. rivularis. In addition, we indicate ERα and Cyp19 as sensitive biomarkers for monitoring of oestrogenic contamination in freshwater environments.

Reproductive effects of oestrogenic endocrine disrupting chemicals in Astyanax rivularis inhabiting headwaters of the Velhas River, Brazil.

The Velhas River is the most polluted river in the state of Minas Gerais, south-eastern Brazil. Due to its historical and environmental relevance, the aim of this study was to evaluate the effects of oestrogenic endocrine disruptors on the reproduction of the lambari Astyanax rivularis, a small-sized species found in headwaters of the São Francisco River basin. Quarterly field samplings were carried out during a reproductive cycle in three streams of the upper Velhas River: S1 (reference site) and S2 and S3 (sites contaminated by untreated sewage). The main oestrogenic compounds were evaluated in water using HPLC/MS. Molecular, histological and reproductive biomarkers were assessed in liver and gonad. The results showed higher average concentrations of oestradiol (>200ng/l) in S2 and S3, oestrone (>250ng/l) in S2 as well as oestriol (>200ng/l), bisphenol A (>190ng/l), and nonylphenol (>600ng/l) in S3 compared to S1 (<70ng/l for all compounds). In S2 and S3, there was an increase in the proportion of females, higher ELISA levels of vitellogenin (Vtg) and proteins of the zona radiata (Zrp) in liver males. Insulin-like growth factor (IGF-I) levels were lower in S2 males, which also had a smaller body size, a smaller seminiferous tubule diameter, a higher proportion of spermatogonia, and lower proportion of spermatozoa in relation to S1. Histopathological analyses detected an increase in yolk deficient oocytes and over-ripening in the contaminated sites, and these alterations were associated to a reduction of hepatic Vtg levels and a delay in spawning, respectively. Intersex specimens with perinucleolar follicles in a multifocal distribution in the testis were detected in S2 and S3. These results indicate that chronic exposure to oestrogenic compounds induced endocrine disruption that may affect wild populations of A. rivularis in the Velhas River.

Influence of a large dam and importance of an undammed tributary on the reproductive ecology of the threatened fish matrinxã Brycon orthotaenia Günther, 1864 (Characiformes: Bryconidae) in southeastern Brazil

A total of 312 adults of matrinxã were captured bimonthly, between 2011-2012, in two sites of the rio São Francisco: immediately downstream of the Três Marias Dam (site 1) and in the confluence with the rio Abaeté (site 2). Specimens of Brycon orthotaenia from site 2 presented higher values of total length (32.24 ± 5.70 cm for females and 26.64 ± 3.79 cm for males) and body weight (506.66 ± 332.17 g for females and 267.36 ± 145.84 g for males), when compared with the site 1. In site 2 was registered the highest GSI means (9.97 ± 2.96 for females and 0.93 ± 0.52 for males), compared with the site 1 means. Reproduction period occurred from October to February. Unlike site 2, no spawned females or spent males were captured in site 1. Significant differences were found between females and males in site 2 (χ2 = 26.84, df = 1, P < 0.05). Physical-chemical parameters of the water presented higher values in site 2. The canonical correlation test indicated that the reproduction of males is less susceptible to environmental factors than that of females. These results show that B. orthotaenia finds favourable conditions to reproduction just in site 2.

Um total de 312 adultos de matrinxã foram capturados bimestralmente entre 2011-2012, em dois pontos do rio São Francisco: imediatamente a jusante da barragem de Três Marias (ponto 1) e na confluência com o rio Abaeté (ponto 2). Exemplares de Brycon orthotaenia do ponto 2 apresentaram maiores valores de comprimento total (32,24 ± 5,70 cm para fêmeas e 26,64 ± 3,79 cm para machos) e peso corporal (506,66 ± 332,17 g para fêmeas e 267,36 ± 145,84 g para machos) quando comparado com peixes do ponto 1. No ponto 2, foi registrado as maiores médias de IGS (9,97 ± 2,96 para fêmeas e 0,93 ± 0,52 para machos) em comparação com o ponto 1. O período reprodutivo ocorreu de outubro a fevereiro. Ao contrário do ponto 2, fêmeas desovadas e machos espermiados não foram capturados no ponto 1. Foram encontradas diferenças significativas entre fêmeas e machos no ponto 2 (χ2 = 26,84, gl = 1, P < 0,05). Parâmetros físico-químicos da água apresentaram maiores valores no ponto 2. A análise de correlação canônica indicou que a reprodução de machos é menos susceptível a fatores ambientais do que de fêmeas. Estes resultados mostram que B. orthotaenia encontra condições favoráveis para a reprodução apenas no ponto 2.

Influence of a large dam and importance of an undammed tributary on the reproductive ecology of the threatened fish matrinxã Brycon orthotaenia Günther, 1864 (Characiformes: Bryconidae) in southeastern Brazil

A total of 312 adults of matrinxã were captured bimonthly, between 2011-2012, in two sites of the rio São Francisco: immediately downstream of the Três Marias Dam (site 1) and in the confluence with the rio Abaeté (site 2). Specimens of Brycon orthotaenia from site 2 presented higher values of total length (32.24 ± 5.70 cm for females and 26.64 ± 3.79 cm for males) and body weight (506.66 ± 332.17 g for females and 267.36 ± 145.84 g for males), when compared with the site 1. In site 2 was registered the highest GSI means (9.97 ± 2.96 for females and 0.93 ± 0.52 for males), compared with the site 1 means. Reproduction period occurred from October to February. Unlike site 2, no spawned females or spent males were captured in site 1. Significant differences were found between females and males in site 2 (χ2 = 26.84, df = 1, P < 0.05). Physical-chemical parameters of the water presented higher values in site 2. The canonical correlation test indicated that the reproduction of males is less susceptible to environmental factors than that of females. These results show that B. orthotaenia finds favourable conditions to reproduction just in site 2.(AU)

Um total de 312 adultos de matrinxã foram capturados bimestralmente entre 2011-2012, em dois pontos do rio São Francisco: imediatamente a jusante da barragem de Três Marias (ponto 1) e na confluência com o rio Abaeté (ponto 2). Exemplares de Brycon orthotaenia do ponto 2 apresentaram maiores valores de comprimento total (32,24 ± 5,70 cm para fêmeas e 26,64 ± 3,79 cm para machos) e peso corporal (506,66 ± 332,17 g para fêmeas e 267,36 ± 145,84 g para machos) quando comparado com peixes do ponto 1. No ponto 2, foi registrado as maiores médias de IGS (9,97 ± 2,96 para fêmeas e 0,93 ± 0,52 para machos) em comparação com o ponto 1. O período reprodutivo ocorreu de outubro a fevereiro. Ao contrário do ponto 2, fêmeas desovadas e machos espermiados não foram capturados no ponto 1. Foram encontradas diferenças significativas entre fêmeas e machos no ponto 2 (χ2 = 26,84, gl = 1, P < 0,05). Parâmetros físico-químicos da água apresentaram maiores valores no ponto 2. A análise de correlação canônica indicou que a reprodução de machos é menos susceptível a fatores ambientais do que de fêmeas. Estes resultados mostram que B. orthotaenia encontra condições favoráveis para a reprodução apenas no ponto 2.(AU)

Oocyte adhesiveness and embryonic development of Astyanax bimaculatus (Linnaeus, 1758) (Pisces: Characidae).

This study shows for the first time the presence of a jelly coat on oocytes of neotropical Characiformes fish. This structure could be responsible for the adhesiveness of Astyanax bimaculatus oocytes, a species widely distributed in South America including in the São Francisco River basin in Brazil. Adult specimens of A. bimaculatus were submitted to artificial reproduction in order to analyse the egg morphology and embryonic development. The eggs were fertilised and kept in incubators with a water temperature of 24°C so that embryogenesis could be monitored. Ovulated and unfertilised oocytes were also collected and submitted to routine histological techniques. Astyanax bimaculatus oocytes were found to be spherical, yellowish, and covered by a thin jelly coat with a slightly adhesive surface. The mean oocyte diameter was 1.03 ± 0.03 mm, the perivitelline space was 0.21 ± 0.02 mm and the jelly coat's thickness was 0.04 ± 0.01 mm. Positive periodic acid-Schiff (PAS) stain and Alcian blue stain pH 2.5 indicated the presence of neutral glycoproteins, and carboxylated acid glycoconjugates on the jelly coat that formed mucosubstances that may be associated with egg adhesiveness. At a water temperature of 24°C, blastopore closure and hatching occurred at 5 h and 17 h after fertilisation, respectively. The results of this study provide essential information for phylogenetic studies and for a better understanding of the reproductive strategy of A. bimaculatus, currently included in the incertae sedis group of the Characidae family due to the lack of monophyly among the families of the group.

Análise da influência do grau de curvatura na ocorrência de desvios apicais após o preparo oscilatório em canais simulados / Analysis of the influence of degree of curvature in the occurrence of apical deviations after the oscillatory preparation in simulated root canals

Introdução e objetivo: Este estudo teve como objetivo analisar a influência do grau de curvatura na ocorrência de desvios apicais após a realização do preparo oscilatório. Material e métodos: Utilizaram-se 20 canais simulados com 21 mm de comprimento, os quais foram divididos em dois grupos experimentais com curvaturas de 20° e 40°. Os canais foram preparados com instrumentos de aço inoxidável acoplados a um contra-ângulo de sistema automatizado de movimentos oscilatórios com redução da velocidade de 10:1. Para a realização da análise do desvio, antes e após o preparo, os canais foram preenchidos com tinta nanquim e fotografados de forma padronizada com o auxílio de uma plataforma. Em seguida, as imagens obtidas foram manipuladas no programa Adobe Photoshop®, sobrepondo a imagem pré e pós-operatória, e com a ferramenta régua fez-se a medição dos desvios a 1 mm do comprimento de trabalho e no meio da área de curvatura. Os dados obtidos foram submetidos à Análise de Variância (Anova) com nível de significância de 5%. Resultados e conclusão: Por meio dos resultados obtidos observou-se que, quanto maior o grau de curvatura, maior foi o desvio. Já em relação ao local de análise, notou-se que na região a 1 mm do comprimento de trabalho houve maior desvio do que no ponto localizado no meio da área de curvatura.

Introduction and objective: This study aims to analyze the influence of degree of curvature in the occurrence of apical deviations after the oscillatory preparation. Material and methods: Twenty simulated root canals of 21 mm in length were divided into two experimental groups according to the degree of curvature: 20° and 40°. The canals were prepared using stainless steel instruments connected to a 10:1 speed reducing contra-angle handpiece with an automatized system of oscillatory movement. When analyzing the deviation, canals were filled with India ink and photographed in a standardized way with the aid of a platform, before and after the accomplishment of the preparation. In sequence, the images obtained were manipulated in Photoshop® Adobe program, preoperative and postoperative images were overlapped, and the deviation was measured with a ruler within 1 mm of working length and in the halfway of curvature. Data were submitted to analysis of variance (Anova) with a significance level of 5%. Results and conclusion: Through the results obtained, it was possible to see that the greater the degree of curvature, the greater the deviation. In relation to the place of analysis, it was possible to realize that there was more deviation in the region of 1 mm of working length than in the halfway of curvature.

Malacoplaquia de bexiga: relato de caso e revisão da literatura / Bladder malacoplakia: case report and literature review

Bladder malacoplakia is a rare granulomatous disease. Although it is a benign disease, it has a recurrent and destructive behavior. Its physiopathogenesis and treatment remain unclear. Grossly, malacoplakia consists of soft yellow colored plaques, and histologicaly it is distinguished by the pathognomonical Michaelis-Gutmann bodies. The authors review the literature and report a case of a 38-year-old patient, with recurrent urinary tract infection, bilateral ureterohydronephrosis and the characteristical vesical trigone plaques. Bilateral percutaneous nephrostomy was performed, followed by transurethral resection of the bladder tumor, thus desobstructing both ureteral orifices and allowing to take away lhe nephrostomies. The diagnosis of bladder malacoplakia was made by histological examination of resected tissue. The disease remains controlled after a therapy with oral antibiotics, bethanecol and vitamin C in afollow-up of 12 months.

Carcinoma sarcomatóide de bexiga: relato de caso / Sarcomatoid carcinoma of the bladder: a case report

Os carcinomas sarcomatóides de bexiga são neoplasias raras, representando apenas 0,3 por cento das patologias malignas da bexiga. Por apresentarem marcado grau de malignidade e agressividade, devem ser sempre consideradas no diagnóstico diferencial dos tumores da bexiga. Os autores revisam a literatura e relatam o caso de um paciente de 62 anos de idade, branco e tabagista, com queixa de hematúria macroscópica, investigada por cistocopia e biópsia. Um exame histológico do tumor sugeriu carcinoma sarcomatóide de bexiga. O diagnóstico foi confirmado por um exame imunohistoquímico. Como o paciente apresentava severa disfunção ventilatória, foi tratado com ressecção transuretral do tumor, recusou a indicação de terapia adjuvante evoluindo para óbito 5 meses após o diagnóstico