PCR Multiplex fluorescente para detecção de bactérias em sêmen bovino

Este estudo teve como objetivo avaliar o limiar de detecção da técnica de PCR multiplex fluorescente aliada a eletroforese capilar na detecção de agentes infecciosos em amostras de sêmen experimentalmente contaminadas com concentrações decrescentes das bactérias Brucella abortus, Leptospira interrogans sorovar pomona, Campylobacter fetus e Haemophilus somnus. Amostras de sêmen bovino foram experimentalmente contaminadas com concentrações decrescentes de bactérias obtidas através de diluições seriadas na base 10 de modo a obter-se amostras contendo desde 1 vez até 10-7 bactérias/mL a partir da concentração inicial de Leptospira pomona, Brucella abortus, Campylobacter fetus e Haemophilus somnus. As diluições foram efetuadas individualmente para cada bactéria, bem como nas diferentes concentrações necessárias para a padronização do teste de multiplex PCR. As extrações de DNA de todas as soluções contendo espermatozóides e bactérias analisadas no presente estudo foram realizadas segundo protocolo descrito por Heinemann et al. (2000). Os produtos de PCR multiplex foram avaliados por eletroforese em gel de poliacrilamida 8% e separação eletroforética por sistema capilar em equipamento automático de análise de fragmentos de DNA MegaBace. Observou-se a amplificação de fragmentos de 193pb, 330pb, 400pb e 415pb a partir do DNA de B. abortus, L. pomona, H. somnus, C. fetus, respectivamente. Na análise por eletroforese capilar de produtos da PCR multiplex do DNA para detecção simultânea dos quatro patógenos observou-se a sinal de positividade até a diluição de 10-3 bactérias/mL vezes da concentração inicial da solução estoque de cada bactéria. A técnica de PCR multiplex aliada à eletroforese capilar foi usada pela primeira vez para o diagnóstico direto de quatro bactérias patogênicas no sêmen, demonstrando ser um método rápido na detecção de bactérias causadoras de doenças reprodutivas.

Effects of Gestational and Lactational Fenvalerate Exposure on Immune and Reproductive Systems of Male Rats

The aim of this study was to determine the consequent reproductive developmental and immunotoxic effects due to exposure to fenvalerate during pregnancy and lactation in male offspring of maternal-treated rats. Pregnant rats were treated daily by oral gavage with 40 or 80 mg/kg of fenvalerate or corn oil (vehicle, control), from d 12 of pregnancy to d 21 of lactation. Immune and reproductive developmental effects were assessed in male offspring at postnatal days (PND) 40 (peripuberty), 60 (postpuberty), and 90 (sexual maturity). Treatment with the higher dose (80 mg/kg) resulted in convulsive behavior, hyperexcitability, and mortality in 45% of the dams. Fenvalerate was detected in the fetus due to placental transfer, as well as in pups due to breast-milk ingestion, persisting in male offspring until PND 40 even though pesticide treatment was terminated on PND 20. However, fenvalerate did not produce marked alterations in age of testicular descent to the scrotum and prepucial separation, parameters indicative of puberty initiation. In contrast, at puberty, there was a reduction in testicular weight and sperm production in male offspring of maternal-treated rats. At adulthood, the sperm counts and fertility did not differ between control and treated groups. Testosterone levels were not changed at any time during reproductive development. Similarly, no apparent exposure-related effects were detected in the histological structures of the lymphohematopoietic system. Data indicate that fenvalerate, in this experimental model, interfered with initial development of the male reproductive system, but that these effects on sperm production or fertility did not persist into adulthood. There was no apparent evidence that fenvalerate altered testosterone levels or produced a disruption in male endocrine functions.

In utero protein restriction causes growth delay and alters sperm parameters in adult male rats

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

Maternal undernutrition and the offspring kidney: from fetal to adult life

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

Rational use of antimicrobials in dentistry during pregnancy

The use of medicines during pregnancy deserves special attention from dentists due to the potential risks to fetal development. The prescription of antimicrobial drugs during this period must be based not only on the etiology of the disease but also on the drug's effect on the embryo, which may be toxic, possibly leading to irreversible lesions. Interest in studies of the teratogenic effects of drugs increased in response to reports of the high incidence of phocomelia in patients treated with thalidomide. Although teratogenicity has long been known, pregnant women today are still exposed to this risk. The effects of drugs depend on the level of susceptibility of the fetus and on the period of exposure during pregnancy. In this context, and considering the paucity of studies on this subject in dentistry, the aim of this review was to offer an up-to-date compilation of data on the antimicrobial drugs most frequently used during pregnancy and the effects of their use.

Efeito do exercício físico contínuo e intervalado no peso e perfil bioquímico de ratas Wistar prenhes e consequências no peso da prole

INTRODUÇÃO: A prática de atividade física é reconhecida como fator importante para a preservação, recuperação e manutenção da saúde. O estímulo à prática de exercícios é crescente, mas quando relacionado à gravidez, dúvidas surgem sobre os efeitos deletérios ou salutares na mãe e no feto. OBJETIVO: O objetivo deste trabalho foi avaliar os efeitos do exercício físico intervalado e contínuo no perfil bioquímico de ratas Wistar prenhes e avaliar o efeito destes exercícios no peso da placenta e dos filhotes. MÉTODOS: Utilizou-se 45 ratas Wistar divididas em grupos de 15 animais segundo o tipo de exercício: controle (GC), exercício contínuo (GCO) e exercício intermitente (GIN). Os exercícios constituíram-se de natação forçada, cinco dias por semana, em piscinas individuais: exercício contínuo (duração de 45 minutos diários com sobrecarga de 5% do peso corporal) e intermitente (45 minutos com estímulos de 15 segundos de exercício e 15 de repouso com sobrecarga de 15% do peso corporal). O exercício foi praticado do primeiro ao 20º dia de prenhez. Após este período avaliou-se o peso e os níveis de glicemia, colesterol total, LDL-C, HDL-C e triglicérides das ratas, assim como o peso da placenta e dos filhotes. RESULTADOS: Não se observou modificação no peso das mães. Houve redução significativa nos níveis de LDL-C. O peso das placentas não variou, mas os pesos dos filhotes variaram estatisticamente entre os três grupos (4,153 ± 0,649; 3,682 ± 0,070 e 3,453 ± 0,052, respectivamente, para os filhos de mães do GC, GIN e GCO). CONCLUSÕES: Conclui-se que a prática do exercício físico contínuo e intermitente por ratas prenhes, neste modelo experimental, não interferiu no peso corpóreo das mesmas, mas interferiu no peso dos filhotes ao nascer

Occurrence of Campylobacter in the genitals of teaser bulls maintained at an embryo transfer center

Em central de transferência de embriões, após os procedimentos de reconhecimento do cio em 37 vacas receptoras, através de quatro rufiões vasectomizados, observou-se que 83% delas apresentavam retorno ao cio e algum corrimento serofibrinoso. Nos exames bacteriológicos realizados nos lavados prepuciais dos rufiões foi isolado o Campylobacter fetus subsp. venerealis em todos, fato que, analisado associadamente com o retorno ao cio das vacas receptoras, é indicativo da ocorrência de campilobacteriose no plantel. Essa ocorrência demonstra a necessidade de medidas eficazes de planejamento de saúde animal, pela utilização de rufiões com desvio lateral do pênis. Uma vez impossibilitado o contato sexual, seria impedida a transmissão do agente durante o coito. Torna-se imperioso consignar que a prática da prevenção racional de enfermidades continua sendo o procedimento mais econômico para uma produtividade animal mais rentável.

Passagem transplacentária e efeitos embriofetais de drogas usadas em anestesia

JUSTIFICATIVA E OBJETIVOS: A anestesia da paciente grávida engloba situações diversas e que devem ser analisadas com muita propriedade. Além da exposição do feto e de uma possível ação tóxica dos agentes a serem utilizados na anestesia, devem ser considerados o período gestacional, as características de cada droga e as doses a serem utilizadas. A falta de informações adequadas sobre o risco do uso de drogas na gestação torna difícil ao anestesiologista uma opção segura quando se vê diante da necessidade de anestesiar uma paciente grávida, tanto para cirurgia não obstétrica, como em cirurgia obstétrica. No primeiro caso, é importante evitar o parto prematuro (ou o aborto) e o aparecimento de alterações permanentes no feto. No segundo caso, não deve haver interferência na contratilidade uterina nem depressão significativa no feto. A finalidade desta revisão é atualizar os conhecimentos sobre a passagem transplacentária e os efeitos maternofetais das drogas usadas em anestesia. CONTEÚDO: São revisados os mecanismos de passagem transplacentária de drogas, os princípios fundamentais de embriofetotoxicidade e analisados alguns aspectos importantes sobre efeitos embriofetais das drogas utilizadas na anestesia. Também é apresentada a classificação de risco teratogênico, de acordo com o FDA, das drogas que o anestesiologista mais utiliza durante o ato anestésico. CONCLUSÕES: Embora ainda persistam muitas dúvidas em relação à escolha de drogas para a anestesia de pacientes grávidas, o anestesiologista dispõe hoje de novas drogas e de informações que lhe permitem oferecer maior segurança para o binômio mãe-feto.

Hemi-ovariossalpingohisterectomia em pacas prenhes e posterior ocorrência de prenhez (Agouti paca, Linnaeus, 1766)

Foi descrita a hemi-ovariossalpingohisterectomia em cinco pacas prenhes mantidas em cativeiro no Setor de Animais Silvestres da Faculdade de Ciências Agrárias e Veterinárias (FCAV-UNESP) de Jaboticabal, São Paulo, Brasil, a fim de observar futura prenhez no corno restante. A tranqüilização foi obtida após aplicação de azaperone (4mg/kg) seguida da aplicação de sulfato de atropina (0,06mg/kg) e da associação de cloridrato de quetamina (20mg/kg) e cloridrato de xilazina (1,5mg/kg), ambos na mesma seringa, para indução da anestesia. A anestesia geral foi obtida mediante inalação de halotano por máscara. Por meio de laparotomia mediana, foram retirados o corno uterino prenhe, o ovário e a tuba uterina, todos do mesmo antímero. Antibióticos (30.000UI/kg de três penicilinas e 12,5mg/kg de duas estreptomicinas) e analgésico (0,02mg/kg de buprenorfina) foram aplicados imediatamente após a cirurgia, sendo repetidos após dois dias. Todas as aplicações foram feitas por via intramuscular. Apesar da permanência de apenas um ovário após a cirurgia, nova prenhez ocorreu no corno restante nas cinco fêmeas submetidas à cirurgia, com o nascimento de filhotes (apenas um por parto) após 215, 248, 276, 302 e 310 dias da hemi-ovariossalpingohisterectomia.

A comparative-analysis of glycosaminoglycans from human umbilical arteries in normal subjects and in pathological conditions affecting pregnancy

BACKGROUND: Vascular cells express different phenotypes in adult and fetal vessels, and the extracellular matrix they synthesize should reflect these differences. Alterations of vascular proteoglycan/glycosaminoglycan is verified in disorders such as hypertension and diabetes, and when occurring during pregnancy, they bring about structural changes to fetal vessels that often lead to impaired fetus growth. Yet there is little data about the extracellular matrix of an important human fetal vessel, the umbilical artery.EXPERIMENTAL DESIGN: This study involved the biochemical characterization of the extracellular matrix of normal umbilical arteries, umbilical arteries from complicated pregnancies (maternal hypertension and diabetes and intrauterine growth retardation syndrome), and, for purpose of comparison, normal adult arteries (aorta and iliac and pulmonary arteries). Although the collagen types I:III ratio was determined in some cases, emphasis was placed on analysis of glycosaminoglycans.RESULTS: Normal umbilical arteries differ from normal adult arteries in that they contain greater concentrations of hyaluronic acid and lesser concentrations of heparan sulfate and chondroitin 4-and 6-sulfate. The umbilical artery also differs from adult arteries in the disaccharide composition of its chondroitin and heparan sulfates and in the molecular weight of this latter glycosaminoglycan. The glycosaminoglycan distribution in umbilical arteries derived from complicated pregnancies is roughly similar to that of controls. However, total glycosaminoglycan and collagen were significantly reduced, and the collagen I:III ratio was increased in the umbilical arteries from hypertension-complicated pregnancies.CONCLUSIONS: the glycosaminoglycan composition of the normal umbilical artery, a fully differentiated tissue, differs in many aspects from that of normal adult arteries. of the cases of complicated pregnancies studied, the extracellular matrix of umbilical arteries was altered only in maternal hypertension. The changes, notably a mild fibrosis, were not very pronounced and should not impair hemodynamic properties of the vessel.

Effects of leucine supplemented diet on intestinal absorption in tumor bearing pregnant rats

Background: It is known that amino acid oxidation is increased in tumor-bearing rat muscles and that leucine is an important ketogenic amino acid that provides energy to the skeletal muscle.Methods: To evaluate the effects of a leucine supplemented diet on the intestinal absorption alterations produced by Walker 256, growing pregnant rats were distributed into six groups. Three pregnant groups received a normal protein diet (18% protein):pregnant (N), tumor-bearing (WN), pair-fed rats (Np). Three other pregnant groups were fed a diet supplemented with 3% leucine (15% protein plus 3% leucine):leucine (L), tumor-bearing (WL) and pair-fed with leucine (Lp). Non pregnant rats (C), which received a normal protein diet, were used as a control group. After 20 days, the animals were submitted to intestinal perfusion to measure leucine, methionine and glucose absorption.Results: Tumor-bearing pregnant rats showed impairment in food intake, body weight gain and muscle protein content, which were less accentuated in WL than in WN rats. These metabolic changes led to reduction in both fetal and tumor development. Leucine absorption slightly increased in WN group. In spite of having a significant decrease in leucine and methionine absorption compared to L, the WL group has shown a higher absorption rate of methionine than WN group, probably due to the ingestion of the leucine supplemented diet inducing this amino acid uptake. Glucose absorption was reduced in both tumor-bearing groups.Conclusions: Leucine supplementation during pregnancy in tumor-bearing rats promoted high leucine absorption, increasing the availability of the amino acid for neoplasic cells and, mainly, for fetus and host utilization. This may have contributed to the better preservation of body weight gain, food intake and muscle protein observed in the supplemented rats in relation to the non-supplemented ones.

Experimental paracoccidioidomycosis in hamsters (Mesocricetus auratus): Gestational interactions

Paracoccidioides brasiliensis is a dimorphic fungus presenting specific steroid hormone receptors, both in the yeast and mycelial forms and estrogen inhibits the transition from mycelium to yeast. In the acute phase, the disease occurs with equal frequency in both sexes but in adults, females are spared. Placental fungal infection has been reported, but references to fetal infection have not been confirmed. We used 78 Syrian female hamsters divided into 3 groups: GI consisted of 30 infected mated females, GII of 20 infected unmated females and GIII of 28 uninfected mated females. Animals of group I were mated 4 weeks after infection and half of them were submitted to cesarean section on day 15 after successful mating; the other half was maintained and submitted to cesarean section and sacrificed 14 weeks after infection. Half of the animals of group II were sacrificed seven weeks and the other half 14 weeks after infection. Uninfected animals of group III were treated the same as the animals of group I. The animals were infected with strain 18 of P. brasiliensis by the intracardiac route. We evaluated the disease by the volume of granulomas in different organs, number of fungi in liver and spleen and the immunologic responses [ELISA, Double Immunodifusion (DID), Delayed Hypersensitivity Skin Test (DHT) and Macrophage Migration Inhibition (MMI)]. We studied the infection through the gestation by evaluation of the abortions, morphologic and clinic examinations of the fetuses. Our results showed that the infection did not transfer to the fetus through the placenta, but the number of abortions was larger among infected females. The newborns of GI females were smaller, weighed less and showed little vitality. The disease was more severe and disseminated in infected mated females, especially in the second sacrifice 14 weeks after inoculation, when the total volume of granulomas in them (56.3 mm) was much greater than in the infected unmated females (12 mm).

Caracterização da fusão caruncular em gestações naturais e de conceptos bovinos clonados

Barreto R.S.N., Miglino M.A., Meirelles F.V., Visintin J.A., Silva S.M., Burioli K.C., Fonseca R., Bertan C., Assis Neto A.C. & Pereira F.T.V. 2009. [Characterization of the caruncular fusion in gestations of natural and cloned bovine conceptuses.] Caracterizacao da fusao caruncular ern gestacoes naturais e de conceptos bovinos clonados. Pesquisa Veterinaria Brasileira 29(10):779-787. Laboratorio de Morfofisiologia da Placenta e Embriao, Faculdade de Zootecnia, Universidade Estadual Paulista, SP294 Km 651, Dracena, SP 17900-000, Brazil. E-mail: fverechia@dracena.unesp.brThe objective of the study was to compare the characteristics of the caruncular fusion in gestations of non-cloned and cloned conceptuses. The non-cloned conceptuses were divided according to the gestation period: Group 1 (2 to 3 months; n=9),II (4 to 6; n=9); III (7 to 8; n=10) and IV (9 n=7). The cloned conceptuses formed the Group V: 9 months; n=4. The caruncles were observed macroscopically (number and dimensions: length, width and height), microscopically and submitted to statistical analysis (5% of significance). We observed three types of macroscopic caruncular fusions: oval (morphologically normal); two united adjacent caruncles and the lobulated type, characterized by regions with several united caruncles presenting a false fusion or deformation of the caruncular parenchyma. The length of the caruncles was 1.55 +/- 0.57; 2.45 +/- 0.55; 4.66 +/- 2.0 and 5.72 +/- 1.90cm for the groups 1, 11, 111, IV respectively. As for the height, the caruncles presented a lineal growth during the gestation: 0.40 +/- 0.15; 0.57 +/- 0.21; 1.0 +/- 0.48 and 1.80 +/- 0.91cm, for the respective groups 1, 11, 111 and IV. The width of the caruncles was similar between the groups I and 11 (0.97 +/- 0.30 e 1.42 +/- 0.71 cm) and the groups III and IV (2.68 +/- 1.22 and 3.52 +/- 1.16cm). When the group V was compared to the IV, the caruncles of the group V presented a larger length (5.72 +/- 1.90 vs. 7.88 +/-.13cm) and width (3.52 +/- 1.16 vs. 4.93 +/- 1.46cm), however they were similar in height (1.80 +/- 0.91 and 2.25 +/- 0.67cm). We verified that in gestations of cloned conceptuses the caruncles presented a larger development than in gestations of non-cloned conceptuses. The fusioned caruncles presented measurements statistically similar to the isolated ones in all the parameters and groups. Under light microscopy, we observed the formation of a stromal axis from the basis of the caruncle to the apex of the fusional fissure, with the histological constitution similar to the endometrial stroma. Three microscopic shapes were also unpublished defined: true fusion with a single axis evident below the fusional fissure; pseudofusion with a double axis in H shape and false fusion with absence of the axis. The first two formats were associated to the oval and lobulated caruncles and the last one to the false fusion with deformation of the caruncle parenchyma. The fusional axis increased in size along the gestation among the groups I, II, III and IV. The group V presented a larger length and width of the axis when compared to the group IV. Thus, in gestations of cloned conceptuses a destruction of the lateral epithelium of the caruncles is associated to an incompetence in the maternal-fetal interdigitation, that compromises the cotyledonary fusion. We suggest that, in gestations derived of cloned conceptuses, the increase of the size of the caruncular fusions is possibly associated to a compensatory mechanism for the metabolic exchanges between mother and fetus, in reason of the smallest number of isolated caruncles.