Incidence and transplacental transmission of Neospora caninum in primiparous females from Bos indicus slaughtered in Presidente Pradente, São Paulo, Brazil

To produce an epidemiological map of neosporosis in Brazil and identify the types of transmission of this disease, the present study evaluated the occurrence of Neospora caninum in Nelore cattle (Bos indicus) in Presidente Prudent, west region of São Paulo state; its vertical transmission; and the early stage in which fetuses are infected. To achieve this, serum samples from 518 slaughtered pregnant heifers and their fetuses were tested by ELISA technique and fetal brain tissues subjected to PCR. One hundred and three heifers (19.88%) had antibodies to N. caninum, as well as 38 (36.8%) of fetuses from 4 months of gestation. The conventional PCR failed to detect N. caninum DNA. These findings show that neosporosis occurs in the area studied and that it may be transmitted the transplacental route, althought N. caninum had not detected in brain tissue from non-aborted fetuses. The use of nested PCR it would be applied to increase the sensitivy of test.

Rabies virus in a pregnant naturally infected southern yellow bat (Lasiurus ega)

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

Structural and ultrastructural characterization of buffalo fetus (Bubalus bubalis) ovarian germinative cells

The objective of the present study was to characterize ovogones, primary oocytes and preantral follicles of buffalo fetus in different ages of gestation. For this, 29 fetuses were collected from a slaughterhouse (Frigol, Brazil) and crown-rump lengths were measured to estimate the fetal age (0-3, 4-6, 7-10 months of gestation). The ovaries were removed and ovarian tissue was processed for classic histology and transmission eletron microscopy examination. The structural evaluation demonstrated that in the first period of the gestation (0-3 months) the buffalo fetus showed ovogones (in mitotic division) and in some cases, the primary oocytes surrounded by somatic cells. In the second period (4-6 months), it was verified that the preantral follicles were completely formed. In the last period (70 month to the end of gestation) the ovaries contained a large amount of preantral follicles, and in some fetuses, antral follicles were observed. The ultrastructural analysis of the ovogones, primary oocytes and preantral follicles showed that these cells have few organelles and the quantity of mitochondria, endoplasmatic reticulum and apparatus Golgi complex is increased as the germinative cells passing from one stage to another.

Efeito do sevofluorano sobre a freqüência cardíaca fetal no terço final de gestação de cadelas

Estudaram-se os efeitos do sevofluorano sobre a freqüência cardíaca nos fetos de cadelas no terço final de gestação, mediante a mensuração da freqüência cardíaca fetal com a utilização do ultra-som. Nove cadelas, sem raça definida, entre um e cinco anos de idade, com aproximadamente 45 dias de gestação, foram anestesiadas com acepromazina (0,05mg/kg, IV), propofol (5mg/kg, IV) e sevofluorano. O monitoramento da freqüência cardíaca fetal foi realizado antes da medicação pré-anestésica (M0), 15 minutos após a intubação traqueal (M1), aos 30 minutos (M2) e 60 minutos (M3) do período de manutenção anestésica. A pressão arterial sistólica (PAS), média (PAM) e diastólica (PAD) foram obtidas pelo método não invasivo, sendo a PAM avaliada também pelo método invasivo. Por meio do monitoramento da freqüência cardíaca fetal média não se observou diferença significativa entre M0, M1 e M2, e verificou-se elevação apenas aos 60 minutos da manutenção anestésica (M3) em relação ao M0, porém sem significado clínico. O protocolo anestésico provocou diminuição significativa da pressão sangüínea arterial materna sem alterar, porém a freqüência cardíaca dos fetos.

Prediction and probability of neonatal outcome in isolated congenital diaphragmatic hernia using multiple ultrasound parameters

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

Fetal Pulmonary Response After Fetoscopic Tracheal Occlusion for Severe Isolated Congenital Diaphragmatic Hernia

OBJECTIVE: To estimate the response in lung growth and vascularity after fetal endoscopic tracheal occlusion for severe congenital diaphragmatic hernia in the prediction of neonatal survival.METHODS: Between January 2006 and December 2010, fetal lung parameters (observed-to-expected lung-to-head ratio; observed-to-expected lung volume; and contralateral lung vascularization index) were evaluated before fetal tracheal occlusion and were evaluated longitudinally every 2 weeks in 72 fetuses with severe isolated congenital diaphragmatic hernia. Thirty-five fetuses underwent fetal endoscopic tracheal occlusion and 37 cases did not.RESULTS: Survival rate was significantly higher in the fetal endoscopic tracheal occlusion group (54.3%) than in the no fetal endoscopic tracheal occlusion group (5.4%, P<.01). Fetal endoscopic tracheal occlusion resulted in a significant improvement in fetal lung size and pulmonary vascularity when compared with fetuses that did not go to the fetal intervention (increase of the observed-to-expected lung-to-head ratio, observed-to-expected total lung volume, and contralateral pulmonary vascularization index 56.2% compared with 0.3%, 37.9% compared with 0.1%, and 98.6% compared with 0.0%, respectively; P<.01). Receiver operating characteristic curves indicated that the observed-to-expected total fetal lung volume was the single best predictor of neonatal survival before fetal endoscopic tracheal occlusion (cutoff 0.23, area under the curve [AUC] 0.88, relative risk 5.3, 95% confidence interval [CI] 1.4-19.7). However, the contralateral lung vascularization index at 4 weeks after fetal endoscopic tracheal occlusion was more accurate in the prediction of neonatal outcome (cutoff 24.0%, AUC 0.98, relative risk 9.9, 95% CI 1.5-66.9) with the combination of observed-to-expected lung volumes and contralateral lung vascularization index at 4 weeks being the best predictor of outcome (AUC 0.98, relative risk 16.6, 95% CI 2.5-112.3).CONCLUSION: Fetal endoscopic tracheal occlusion improves survival rate by increasing the lung size and pulmonary vascularity in fetuses with severe congenital diaphragmatic hernia. The pulmonary response after fetal endoscopic tracheal occlusion can be used to predict neonatal survival. (Obstet Gynecol 2012; 119: 93-101) DOI: 10.1097/AOG.0b013e31823d3aea

Three-Dimensional Ultrasonographic Assessment of Fetal Total Lung Volume as a Prognostic Factor in Primary Pleural Effusion

Objectives-The purpose of this study was to predict perinatal outcomes using fetal total lung volumes assessed by 3-dimensional ultrasonography (3DUS) in primary pleural effusion.Methods-Between July 2005 and July 2010, total lung volumes were prospectively estimated in fetuses with primary pleural effusion by 3DUS using virtual organ computer-aided analysis software. The first and last US examinations were considered in the analysis. The observed/expected total lung volumes were calculated. Main outcomes were perinatal death (up to 28 days of life) and respiratory morbidity (orotracheal intubation with mechanical respiratory support >48 hours).Results-Twelve of 19 fetuses (63.2%) survived. Among the survivors, 7 (58.3%) had severe respiratory morbidity. The observed/expected total lung volume at the last US examination before birth was significantly associated with perinatal death (P < .01) and respiratory morbidity (P < .01) as well as fetal hydrops (P < .01) and bilateral effusion (P = .01).Conclusions-Fetal total lung volumes may be useful for the prediction of perinatal outcomes in primary pleural effusion.

Expression of fibroblast growth factor-8 and its cognate receptors, fibroblast growth factor receptor (FGFR)-3c and-4, in fetal bovine preantral follicles

Paracrine cell signaling is thought to be important for ovarian follicle development, and a role for some members of the fibroblast growth factor (FGF) family have been suggested. In the present study, we tested the hypothesis that FGF-8 and its cognate receptors (FGFR-3c and FGFR-4) are expressed in bovine preantral follicles. Reverse transcription-polymerase chain reaction was used to amplify bovine FGF-8, FGFR-3c, and FGFR-4 from preantral follicle samples and a variety of fetal and adult tissues. All three genes were widely expressed in fetal tissues, with a restricted expression pattern in adult tissues. FGF-8 and FGFR-3c were expressed in secondary follicles in 70% of fetuses examined, whereas FGFR-4 expression was significantly less frequent (20%). FGFR-3c expression frequency was significantly lower in primordial compared to secondary follicles, and FGF-8 expression showed a similar trend. FGFR-4 was only observed when all follicle classes of an individual were expressing both FGF-8 and FGFR-3c. We conclude that FGF-8 and its receptors are expressed in preantral follicles in a developmentally regulated manner. (C) 2005 Wiley-Liss, Inc.

The toxic effects of Coleus barbatus B. on the different periods of pregnancy in rats

Extracts of Coleus bal barbatus B. have been used in folk medicine to interrupt pregnancy. In order to evaluate if this plant interferes with embryo implantation or with the normal development of the concepts, pregnant Wistar rats were treated with increasing doses (220, 440 and 880 mg/kg per day) of a hydroalcoholic extract of C. barbatus. The rats received the extract by gavage from days 0 to 5 of pregnancy (preimplantation period) or 6 to 15 (organogenic period). Control groups received distilled water during the same periods: the animals were killed at term for the evaluation of maternal and fetal parameters. The results showed that the treatment with 880 mg/kg per day of the extract of C. barbatus before embryo implantation caused delayed fetal development and an anti-implantation effect, which justifies the popular use of this extract with abortive purposes. After embryo implantation delayed development associated with maternal toxicity was observed in the fetuses of the group which received 880 mg/kg per day. (C) 2000 Elsevier B.V. Ireland Ltd. All rights reserved.

Somatic cell nuclear transfer is associated with altered expression of angiogenic factor systems in bovine placentomes at term

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

Fetotoxicity caused by the interaction between zinc and arsenic in mice

Arsenic is an environmental pollutant that induces congenital malformations in experimental models and can contribute to human birth defects. The environmental exposure to arsenic is relatively small when compared with the doses required to cause teratogenicity in mice and other laboratory animals. In order to study the action of zinc in the arsenic-induced teratogenicity, in the present work mice were either pretreated with zinc and later with arsenic or were treated simultaneously with zinc and arsenic in vivo and in vitro. Following administration of arsenate on gestation day 8, pregnant females were killed on the 17th day of gestation; maternal and fetal data were collected by laparotomy and used to calculate reproductive parameters. Fetuses were analyzed for the presence of external malformation and, after the appropriate processing, visceral and skeletal analyses were accomplished. Conceptuses were exposed in whole embryo culture to arsenicals on gestation day 8 (3-6 somite stage). After a 26 h culture period, morphological development was assessed. Neither pretreatment with zinc nor simultaneous administration of zinc prevented arsenic teratogenicity in these experimental models. (C) 2002 Wiley-Liss, Inc.

Reproductive effects in male rats exposed to diuron

Diuron is a ureic herbicide considered to have very low toxicity. The present study evaluated several aspects of reproductive toxicity of diuron in adult male rats. Diuron was diluted in corn oil and administered by oral gavage to groups of 18-20 rats at doses of 0, 125 or 250 mg/kg per day for 30 days; the control group received only the corn oil vehicle. At the end of the treatment period, approximately half the animals from each group were assigned to one of two terminal assessment lines: (1) reproductive organ, liver and kidney weights; measurement of diuron concentrations in liver and kidney; plasma testosterone determinations; evaluation of daily sperm production per testis; sperm number and sperm transit time in the epididymis; or (2) sexual behavior assessment during cohabitation with a receptive female; fertility and pregnancy outcome after natural mating; testicular, epididymal, kidney and liver histopathology; sperm morphology. After 30 days of oral diuron treatment, there were no treatment-related changes in body weights, but dose-related diuron residues were detected in the liver of all treated rats and absolute and relative liver weights were increased in both groups. There were no statistically significant differences between the treated and control groups obtained in plasma testosterone concentrations, or in parameters of daily sperm production, sperm reserves in the epididymis, sperm morphology or measured components of male sexual behavior. on the other hand, the number of fetuses in the litters from diuron-treated rats was slightly smaller than litters from control rats. Therefore, although the results did not indicate that diuron exposure resulted in direct male reproductive toxicity in the rat, they suggest that additional studies should be undertaken to investigate the possible effects on fertility and reproductive performance. (c) 2006 Elsevier B.V. All rights reserved.

Functional and Morphological Reproductive Aspects in Male Rats Exposed to Di-n-Butyl Phthalate (DBP) in Utero and During Lactation

The potential adverse reproductive effects, with emphasis on the epididymis, of in utero and lactational exposure to 100 mg/kg/d di-n-butyl phthalate (DBP) in adult male rat offspring were investigated. The fetal testis histopathology was also determined. The selected endpoints included reproductive organ weights, sperm motility and morphology, sperm epididymal transit time, sperm quantity in the testis and epididymis, hormonal status, fetal testis and epididymal histopathology and stereology, and androgen receptor (AR), aquaporin 9 (AQP9), and Ki-67 immunoreactivities. Pregnant females were divided into two groups: control (C) and treated (T). The treated females received DBP (100 mg/kg/d, by gavage) from gestation day (GD) 12 to postnatal day (PND) 21, while control dams received the vehicle. Some pregnant dams were killed by decapitation on GD20, and testes from male fetuses were collected for histopathogy. Male rats from other dams were killed at PND 90. Fetal testes from treated group showed Leydig-cell clusters, presence of multinucleated germinative cells, and increase of the interstitial component. Testosterone levels and reproductive organ weights were similar between the treated and control adult groups. DBP treatment did not markedly affect relative proportions of epithelial, stromal, or luminal compartments in the epididymis; sperm counts in the testis and epididymis; sperm transit time; or sperm morphology and motility in adult rats. The AR and AQP9 immunoreactivities and proliferation index were similar for the two groups. These results showed that fetal testes were affected by DBP as evidenced by testicular histopathologic alterations, but reproductive parameters and epididymal structure/function were not significantly altered in the adult animals exposed to 100 mg/kg DBP in utero and during lactation.

Development of the insulin secretion mechanism in fetal and neonatal rat pancreatic B-cells: response to glucose, K+, theophylline, and carbamylcholine

We studied the development of the insulin secretion mechanism in the pancreas of fetal (19- and 21-day-old), neonatal (3-day-old), and adult (90-day-old) rats in response to stimulation with 8.3 or 16.7 mM glucose, 30 mM K+, 5 mM theophylline (Theo) and 200 µM carbamylcholine (Cch). No effect of glucose or high K+ was observed on the pancreas from 19-day-old fetuses, whereas Theo and Cch significantly increased insulin secretion at this age (82 and 127% above basal levels, respectively). High K+ also failed to alter the insulin secretion in the pancreas from 21-day-old fetuses, whereas 8.3 mM and 16.7 mM glucose significantly stimulated insulin release by 41 and 54% above basal levels, respectively. Similar results were obtained with Theo and Cch. A more marked effect of glucose on insulin secretion was observed in the pancreas of 3-day-old rats, reaching 84 and 179% above basal levels with 8.3 mM and 16.7 mM glucose, respectively. At this age, both Theo and Cch increased insulin secretion to close to two-times basal levels. In islets from adult rats, 8.3 mM and 16.7 mM glucose, Theo, and Cch increased the insulin release by 104, 193, 318 and 396% above basal levels, respectively. These data indicate that pancreatic B-cells from 19-day-old fetuses were already sensitive to stimuli that use either cAMP or IP3 and DAG as second messengers, but insensitive to stimuli such as glucose and high K+ that induce membrane depolarization. The greater effect of glucose on insulin secretion during the neonatal period indicates that this period is crucial for the maturation of the glucose-sensing mechanism in B-cells.

Fertility of male adult rats submitted to forced swimming stress

We investigated whether stress interferes with fertility during adulthood. Male Wistar rats (weighing 220 g in the beginning of the experiment) were forced to swim for 3 min in water at 32ºC daily for 15 days. Stress was assessed by the hot-plate test after the last stressing session. To assess fertility, control and stressed males (N = 15 per group) were mated with sexually mature normal females. Males were sacrificed after copulation. Stress caused by forced swimming was demonstrated by a significant increase in the latency of the pain response in the hot-plate test (14.6 ± 1.25 s for control males vs 26.0 ± 1.53 s for stressed males, P = 0.0004). No changes were observed in body weight, testicular weight, seminal vesicle weight, ventral prostate weight or gross histological features of the testes of stressed males. Similarly, no changes were observed in fertility rate, measured by counting live fetuses in the uterus of normal females mated with control and stressed males; no dead or incompletely developed fetuses were observed in the uterus of either group. In contrast, there was a statistically significant decrease in spermatid production demonstrated by histometric evaluation (154.96 ± 5.41 vs 127.02 ± 3.95 spermatids per tubular section for control and stressed rats, respectively, P = 0.001). These data demonstrate that 15 days of forced swimming stress applied to adult male rats did not impair fertility, but significantly decreased spermatid production. This suggests that the effect of stress on fertility should not be assessed before at least the time required for one cycle of spermatogenesis.