Quantificação do líquido amniótico pela ultra-sonografia tridimensional e bidimensional no primeiro trimestre da gestação

OBJETIVO: quantificar os valores do líquido amniótico no primeiro trimestre da gestação, em fetos normais, por meio de ultra-sonografia tridimensional e bidimensional. MÉTODOS: foram avaliados 25 fetos normais, da 8ª à 11ª semana de gestação. O estudo foi do tipo prospectivo longitudinal. As medidas do volume de líquido amniótico foram obtidas por meio de ultra-sonografia endovaginal, modos tridimensional e bidimensional. O estudo bidimensional consistiu em determinar o volume por cálculo matemático com base na forma de um elipsóide (constante 0,52), obtendo-se o volume do saco amniótico e do embrião. No estudo tridimensional, o volume do líquido amniótico foi feito pela técnica VOCAL, utilizando os graus de rotação 6, 9, 15 e 30°. Foi considerado como resultado final o volume do líquido amniótico obtido pelo grau de rotação6. Em ambos, o volume de líquido amniótico foi obtido pela subtração do volume do saco amniótico pela medida volumétrica do embrião. Para análise estatística utilizamos análise de variância (ANOVA), correlação e análise de regressão. O nível de significância adotado foi p < 0,05. RESULTADOS: a evolução no volume de líquido amniótico pela ultra-sonografia bidimensional foi de 5,4 para 39,5 cm³ da 8ª para a 11ª semana (ANOVA - p < 0,05). Observamos correlação entre idade gestacional e volume de líquido amniótico (p < 0,001, r² = 88,3%). No estudo tridimensional o volume de líquido amniótico aumentou de 5,7 para 42,9 cm³ da 8ª para a 11ª semana (ANOVA - p < 0,05). Também observamos correlação entre idade gestacional e volume de líquido amniótico (p < 0,001, r² = 98,1%). CONCLUSÃO: há aumento no volume de líquido amniótico no primeiro trimestre da gestação, quando avaliado nos modos bidimensional e tridimensional. Além disso, demonstramos que, quanto maior a idade gestacional, maior é o volume de líquido amniótico.

Uropatias obstrutivas bilaterais fetais: sinais ultrassonográficos durante a gravidez e evolução pós-natal

OBJETIVO: verificar a associação entre sinais ultrassonográficos durante a gestação e evoluções pós-natais em casos de fetos com uropatias obstrutivas bilaterais, acompanhados de forma expectante. MÉTODOS: fetos com uropatias obstrutivas bilaterais apresentando oligoâmnio grave e tórax estreito foram comparados a fetos com uropatias obstrutivas bilaterais que não desenvolveram estas alterações com relação à presença ou ausência de cistos em ambos os rins e à presença ou ausência de hiperecogenicidade de parênquima em ambos os rins. Casos em que houve óbito do neonato foram comparados com aqueles em que o neonato teve alta do berçário em relação aos mesmos aspectos ecográficos renais acima citados, à presença de oligoâmnio grave e de tórax estreito. A sensibilidade, a especificidade, os valores preditivos positivo e negativo da presença de cistos renais bilaterais, hiperecogenicidade renal bilateral, oligoâmnio grave e tórax fetal estreito para óbito do neonato foram calculados. RESULTADOS: o oligoâmnio grave e o tórax estreito foram mais frequentes (p=0,03; p<0,001) nos fetos que tiveram cistos renais bilaterais quando comparados àqueles com parênquimas renais ecograficamente normais. O óbito neonatal foi mais frequente entre os casos que tiveram oligoâmnio grave (p<0,001), tórax estreito (p<0,001) e cistos renais bilaterais (p<0,002) quando respectivamente comparados aos casos sem essas alterações. Os melhores valores de sensibilidade, especificidade, valores preditivos positivo e negativo para óbito do neonato/lactente foram obtidos com o uso do aspecto ecográfico tórax estreito, tendo sido de 81,8, 100, 100 e 79,3%, respectivamente. CONCLUSÕES: Em casos de fetos com uropatias obstrutivas bilaterais acompanhados de forma expectante, os sinais ultrassonográficos mais associados ao mau prognóstico são o oligoâmnio grave, o tórax fetal estreito e a presença de cistos renais bilaterais.

Avaliação da adaptação psicossocial na gravidez em gestantes brasileiras

RESUMO OBJETIVO: avaliar a adaptação psicossocial na gravidez, por intermédio da tradução e adaptação de instrumento específico, para ser usado em gestantes brasileiras. MÉTODOS: estudo observacional de corte transversal. Foi realizada a tradução e adaptação transcultural do PSeQ (Prenatal Self-evaluation Questionnaire) seguindo todas as etapas metodológicas exigidas. aplicou-se um questionário contendo perguntas abertas e fechadas de forma a caracterizar os dados sócio-demográficos e clínicos das gestantes (n=36). a análise estatística constou de média, desvio padrão (DP), freqüência absoluta e relativa. Para análise da consistência interna utilizou-se o coeficiente alfa de Cronbach, por meio do SPSS versão 17.0. RESULTADOS: as voluntárias apresentaram baixo nível sócio-econômico, média de idade de 25,1 anos ( 5,5), idade gestacional média de 25,9 semanas ( 8,1). Destas, 58,3% não haviam planejado a atual gravidez. O pré-teste mostrou que 75% das gestantes consideraram o questionário de fácil entendimento. Quanto ao instrumento PSEQ, a identificação com o papel materno foi a sub-escala que apresentou maior média 24,8 ( 5,6), enquanto o relacionamento com a mãe apresentou a menor média 15,4 ( 7,7). a consistência interna variou entre 0,52-0,89. CONCLUSÃO:a avaliação psicossocial materna no pré-natal mostra-se importante no acompanhamento da progressão da gestação e permite a intervenção mediante ações de promoção e prevenção no bem-estar materno-infantil.

Hofbauer cells morphology and density in placentas from normal and pathological gestations

PURPOSE: In placentas from uncomplicated pregnancies, Hofbauer cells either disappear or become scanty after the fourth to fifth month of gestation. Immunohistochemistry though, reveals that a high percentage of stromal cells belong to Hofbauer cells. The aim of this study was to investigate the changes in morphology and density of Hofbauer cells in placentas from normal and pathological pregnancies. METHODS: Seventy placentas were examined: 16 specimens from normal term pregnancies, 10 from first trimester's miscarriages, 26 from cases diagnosed with chromosomal abnormality of the fetus, and placental tissue specimens complicated with intrauterine growth restriction (eight) or gestational diabetes mellitus (10). A histological study of hematoxylin-eosin (HE) sections was performed and immunohistochemical study was performed using the markers: CD 68, Lysozyme, A1 Antichymotrypsine, CK-7, vimentin, and Ki-67. RESULTS: In normal term pregnancies, HE study revealed Hofbauer cells in 37.5% of cases while immunohistochemistry revealed in 87.5% of cases. In first trimester's miscarriages and in cases with prenatal diagnosis of fetal chromosomal abnormalities, both basic and immunohistochemical study were positive for Hofbauer cells. In pregnancies complicated with intrauterine growth restriction or gestational diabetes mellitus, a positive immunoreaction was observed in 100 and 70% of cases, respectively. CONCLUSIONS: Hofbauer cells are present in placental villi during pregnancy, but with progressively reducing density. The most specific marker for their detection seems to be A1 Antichymotrypsine. It is remarkable that no mitotic activity of Hofbauer cells was noticed in our study, as the marker of cellular multiplication Ki-67 was negative in all examined specimens.

Does hyperglycemia in pregnancy change fetal kidney growth?: a longitudinal prospective study

PURPOSE: To measure fetal renal volume in normoglycemic and hyperglycemic pregnancies. METHODS: A longitudinal prospective study was conducted and included 92 hyperglycemic and 339 normoglycemic pregnant women attended at the prenatal service of a hospital from Rio de Janeiro State. Ultrasound examinations were performed to estimate gestational age at baseline and the kidney volume was estimated using the prolate ellipsoid volume equation. RESULTS: Fetal kidney volume growth between normoglycemic and hyperglycemic pregnancies are significantly different. The fetal kidney volume growth in pregnancy is positively correlated with gestational age explained by these predictor equations, by group: normal renal volume = exp (6.186+0.09×gestational week); hyperglycemic renal volume = exp (6.978+0.071×gestational week) and an excessive growth pattern for hyperglycemic pregnancies may be established according to gestational age. CONCLUSION: This is important for early detection of abnormalities in pregnancy, particularly in diabetic mothers.

Triplet pregnancies: perinatal outcome evolution

PURPOSE: To evaluate the obstetric and perinatal outcomes evolution of triplet pregnancies.METHODS: A prospective observational study was conducted in triplet pregnancies delivered over 16 years in a tertiary obstetric center with differentiated perinatal support. Evaluation of demographic factors, obstetric complications, gestational age at delivery, mode of delivery, birth weight and immediate newborn outcome were done over a 16 years period. A global characterization of the sample was performed considering the listed parameters. Variables were categorized in three groups according to year of occurrence: 1996-2000, 2001-2006, 2007-2011, and all parameters were compared.RESULTS: Of the 33 triplets included, 72.7% resulted from induced pregnancies. All except one patient received prenatal corticosteroids and five received tocolytics. All women delivered prenatally and no significant differences were seen in the mean gestational age at delivery or birth weight towards time. There were three intrauterine fetal deaths. Neonatal immediate outcomes were not significantly different over the years.CONCLUSION: Despite remarkable progresses in perinatal and neonatal cares, no noticeable impact in triplet gestations' outcomes was seen, sustaining that triplets should be avoided due to their great risk of prematurity and neonatal morbidities, either by limiting the numbers of embryos transferred or by fetal reduction.

Pregnancy in women undergoing hemodialysis: case series in a Southeast Brazilian reference center

PURPOSE: To describe maternal and neonatal outcomes in pregnant women undergoing hemodialysis in a referral center in Brazilian Southeast side.METHODS: Retrospective and descriptive study, with chart review of all pregnancies undergoing hemodialysis that were followed-up at an outpatient clinic of high- risk prenatal care in Southeast Brazil.RESULTS: Among the 16 women identified, 2 were excluded due to follow-up loss. In 14 women described, hypertension was the most frequent cause of chronic renal failure (half of cases). The majority (71.4%) had performed hemodialysis treatment for more than one year and all of them underwent 5 to 6 hemodialysis sessions per week. Eleven participants had chronic hypertension, 1 of which was also diabetic, and 6 of them were smokers. Regarding pregnancy complications, 1 of the hypertensive women developed malignant hypertension (with fetal growth restriction and preterm delivery at 29 weeks), 2 had acute pulmonary edema and 2 had abruption placenta. The mode of delivery was cesarean section in 9 women (64.3%). All neonates had Apgar score at five minutes above 7.CONCLUSIONS: To improve perinatal and maternal outcomes of women undergoing hemodialysis, it is important to ensure multidisciplinary approach in referral center, strict control of serum urea, hemoglobin and maternal blood pressure, as well as close monitoring of fetal well-being and maternal morbidities. Another important strategy is suitable guidance for contraception in these women.

Medico-legal litigation in Obstetrics: a characterization analysis of a decade in Portugal

PURPOSE: It was to analyse the most critical areas in Obstetrics and to suggest measures to reduce or avoid the situations most often involved in these disputes. METHODS: Obstetrics cases submitted to the Medico-legal Council since the creation of the National Institute of Legal Medicine and Forensic Sciences in 2001 until 2011 were evaluated. A comprehensive characterization, determination of absolute/relative frequencies, hypothesis of a linear trend over the years and the association between each parameter was done. RESULTS: The analysis has shown no significantly linear trend. The most common reasons for disputes were perinatal asphyxia (50%), traumatic injuries of the newborn (24%), maternal sequelae (19%) and issues related to prenatal diagnosis and/or obstetric ultrasound (5.4%). Perinatal asphyxia showed no significantly linear trend (p=0.58) and was usually related to perinatal deaths or permanent neurologic sequelae in newborn children. Traumatic injuries of the newborn, mostly related to instrumented deliveries, shoulder dystocia or vaginal delivery in breech presentation, has shown a significantly increased linear trend (p<0.001), especially related to instrumented deliveries. The delay/absence of cesarean section was the clinical procedure questioned in a significantly higher number of cases of perinatal asphyxia (68.7%) and of traumatic lesions of the newborn due to instrumented deliveries (20.5%). CONCLUSION: It is important to improve and correct theoretical/practical daily clinical performance in these highlighted areas, in order to reduce or even avoid situations that could end up in medico-legal litigations.

Reproductive outcome in pregnant women with recurrent pregnancy loss

Abstract PURPOSE: To estimate the future pregnancy success rate in women with a history of recurrent pregnancy loss. METHODS: A retrospective cohort study including 103 women seen at a clinic for recurrent pregnancy loss (loss group) between January 2006 and December 2010 and a control group including 204 pregnant women seen at a low-risk prenatal care unit between May 2007 and April 2008. Both groups were seen in the university teaching hospital the Maternidade Climério de Oliveira, Salvador, Bahia, Brazil. Reproductive success rate was defined as an alive-birth, independent of gestational age at birth and survival after the neonatal period. Continuous variables Means and standard deviations (SD) were compared using Student's t-test and nominal variables proportions by Pearson χ2test. RESULTS: Out of 90 who conceived, 83 (91.2%) had reproductive success rate. There were more full-term pregnancies in the control than in the loss group (174/187; 92.1 versus 51/90; 56.7%; p<0.01). The prenatal visits number was satisfactory for 76 (85.4%) women in the loss group and 125 (61.3%) in the control (p<0.01). In this, the beginning of prenatal care was earlier (13.3; 4.2 versus 19.6; 6.9 weeks). During pregnancy, the loss group women increased the weight more than those in the control group (58.1 versus 46.6%; p=0.04). Although cervix cerclage was performed in 32/90 women in the loss group, the pregnancy duration mean was smaller (34.8 weeks; SD=5.6 versus 39.3 weeks; SD=1.6; p<0.01) than in the control group. Due to gestational complications, cesarean delivery predominated in the loss group (55/83; 64.7 versus 73/183; 39.5%; p<0.01). CONCLUSION: A very good reproductive success rate can be attributed to greater availability of healthcare services to receive pregnant women, through prenatal visits (scheduled or not), cervical cerclage performed on time, and available hospital care for the mother and newborn.

Características morfométricas do desenvolvimento do timo em cães

O timo é um órgão de grande importância imunológica durante a vida fetal e o período neonatal. Ele é o precursor da linfopoiese e apresenta alta atividade linfopoiética, sendo também o maior órgão linfóide durante a vida intrauterina e até o nascimento. Neste estudo, os aspectoso morfológicos do desenvolvimento prenatal de timos de cães foram descritos em fetos de diferentes idades e sexos. Vinte e quatro fetos de cães domésticos, sem raça definida, machos e fêmeas, foram divididos em 4 grupos etários. O timo apresentou-se composto por dois lobos unidos por um tecido de conexão, com coloração rósea, localizados no espaço mediastinal cranial. A porção cranial estendeu-se pouco além do primeiro par de costelas. As células que formavam o parênquima do timo estavam bem organizadas. Agrega dos concêntricos chamados Corpúsculos de Hassal foram observados, envolvidos por uma delgada cápsula de tecido conjuntivo que é espessada por tecido adiposo. O tamanho dos vasos aumentou gradativamente com a idade dos fetos, sendo maior nos animais próximos ao nascimento. Os timos das fêmeas apresentaram maior volume (Vref) e dimensões de tamanho que nos machos.

Fetal alcohol spectrum disorders in Finnish children and adolescents : diagnosis, cognition, behavior, adaptation and brain metabolic alterations

Alcohol consumption during pregnancy can potentially affect the developing fetus in devastating ways, leading to a range of physical, neurological, and behavioral alterations most accurately termed Fetal Alcohol Spectrum Disorders (FASD). Despite the fact that it is a preventable disorder, prenatal alcohol exposure today constitutes a leading cause of intellectual disability in the Western world. In Western countries where prevalence studies have been performed the rates of FASD exceed, for example, autism spectrum disorders, Down’s syndrome and cerebral palsy. In addition to the direct effects of alcohol, children and adolescents with FASD are often exposed to a double burden in life, as their neurological sequelae are accompanied by adverse living surroundings exposing them to further environmental risk. However, children with FASD today remain remarkably underdiagnosed by the health care system. This thesis forms part of a larger multinational research project, The Collaborative Initiative on Fetal Alcohol Spectrum Disorders (the CIFASD), initiated by the National Institute of Alcohol Abuse and Alcoholism (NIAAA) in the U.S.A. The general aim of the present thesis was to examine a cohort of children and adolescents growing up with fetal alcohol-related damage in Finland. The thesis consists of five studies with a broad focus on diagnosis, cognition, behavior, adaptation and brain metabolic alterations in children and adolescents with FASD. The participants consisted of four different groups: one group with histories of prenatal exposure to alcohol, the FASD group; one IQ matched contrast group mostly consisting of children with specific learning disorder (SLD); and two typically-developing control groups (CON1 and CON2). Participants were identified through medical records, random sampling from the Finnish national population registry and email alerts to students. Importantly, the participants in the present studies comprise a group of very carefully clinically characterized children with FASD as the studies were performed in close collaboration with leading experts in the field (Prof. Edward Riley and Prof. Sarah Mattson, Center for Behavioral Teratology, San Diego State University, U.S.A; Prof. Eugene Hoyme, Sanford School of Medicine, University of South Dakota, U.S.A.). In the present thesis, the revised Institute of Medicine diagnostic criteria for FASD were tested on a Finnish population and found to be a reliable tool for differentiating among the subgroups of FASD. A weighted dysmorphology scoring system proved to be a valuable additional adjunct in quantification of growth deficits and dysmorphic features in children with FASD (Study 1). The purpose of Study 2 was to clarify the relationship between alcohol-related dysmorphic features and general cognitive capacity. Results showed a significant correlation between dysmorphic features and cognitive capacity, suggesting that children with more severe growth deficiency and dysmorphic features have more cognitive limitations. This association was, however, only moderate, indicating that physical markers and cognitive capacity not always go hand in hand in individuals with FASD. Behavioral problems in the FASD group proved substantial compared to the typically developing control group. In Study 3 risk and protective factors associated with behavioral problems in the FASD group were explored further focusing on diagnostic and environmental factors. Two groups with elevated risks for behavioral problems emerged: length of time spent in residential care and a low dysmorphology score proved to be the most pervasive risk factor for behavioral problems. The results underscore the clinical importance of appropriate services and care for less visibly alcohol affected children and highlight the need to attend to children with FASD being raised in institutions. With their background of early biological and psychological impairment compounded with less opportunity for a close and continuous caregiver relationship, such children seem to run an especially great risk of adverse life outcomes. Study 4 focused on adaptive abilities such as communication, daily living skills and social skills, in other words skills that are important for gradually enabling an independent life, maintain social relationships and allow the individual to become integrated into society. The results showed that adaptive abilities of children and adolescents growing up with FASD were significantly compromised compared to both typically-developing peers and IQ-matched children with SLD. Clearly different adaptive profiles were revealed where the FASD group performed worse than the SLD group, who in turn performed worse than the CON1 group. Importantly, the SLD group outperformed the FASD group on adaptive behavior in spite of comparable cognitive levels. This is the first study to compare adaptive abilities in a group of children and adolescents with FASD relative to both a contrast group of IQ-matched children with SLD and to a group of typically-developing peers. Finally, in Study 5, through magnetic resonance spectroscopic imaging (MRS) evidence of longstanding neurochemical alterations were observed in adolescents and young adults with FASD related to alcohol exposure in utero 14-20 years earlier. Neurochemical alterations were seen in several brain areas: in frontal and parietal cortices, corpus callosum, thalamus and frontal white matter areas as well as in the cerebellar dentate nucleus. The findings are compatible with neuropsychological findings in FASD. Glial cells seemed to be more affected than neurons. In conclusion, more societal efforts and resources should be focused on recognizing and diagnosing FASD, and supporting subgroups with elevated risk of poor outcome. Without adequate intervention children and adolescents with FASD run a great risk of marginalization and social maladjustment, costly not only to society but also to the lives of the many young people with FASD.

Smoking during pregnancy and fetal brain development

Background: Although the knowledge of adverse effects of smoking during pregnancy has increased in recent years, more research is needed to gain a better understanding of the effects of smoking during pregnancy. Smoking exposure is the most common preventable factor that causes adverse pregnancy outcomes. Aims and Methods: First, data on smoking habits during pregnancy from the Nordic Medical Birth Registers was used to study the national differences in trends of smoking during pregnancy. Second, the effects of prenatal smoking exposure on fetal brain development, assessed by brain MRI at term age, were studied by using data from the multidisciplinary PIPARI Study consisting of a 6-year cohort of VLBW/VLGA infants (n = 232, of which 18.1% were exposed to prenatal smoking) born in Turku University Hospital, Finland. Third, the effects of prenatal smoking exposure on psychiatric morbidity and use of psychotropic medication were studied in a cohort of children born from 1987–1989 in Finland (n = 175,869, of which 15.3% were exposed). The data used were obtained from population-based longitudinal registers from the National Institute of Health and Welfare, the Statistics Finland, and the Finnish Social Insurance Institution. Results: Smoking rates during pregnancy differed considerably between the countries. Smoking rates were highest among teenagers and women with lower socioeconomic positions. The smoking prevalence was found to be increasing among teenagers in both Finland and Norway. Prenatal smoking exposure was associated with smaller frontal lobe and cerebellar volumes in preterm infants. A clear association was found between prenatal smoking exposure and psychiatric morbidity treated with specialized hospital care and the use of various psychotropic medications. Conclusions: Prenatal smoking exposure had adverse effects on fetal brain development. These effects might explain part of the association found between smoking exposure and psychiatric problems in later life. Our study suggests that prenatal smoking exposure is linked with both mild and severe psychiatric problems. This study emphasizes the importance of efforts to reduce smoking during pregnancy.

Chorion biopsy in mongrel dogs

With the great development of the gestational studies in all of the species, we noticed the necessity of adaptations of these techniques for prenatal diagnosis in dogs. Based on this, we studied the feasibility of chorion biopsy guided by ultrasound. Our results demonstrated accuracy on the sex determination being 2 males and 12 females, as well as it would be possible to identify chromosome alteration due to the quality of samplings. Sex determination was accomplished with the identification of Y gene chromosomes in PCR technique. After the collection, fragments were prepared for light microscopy studies and revealed fetal chorion tissue, blood colloid and erythrocyte. In the whole material we found hemosiderin impregnations due to the hemolysis and to the residue of blood of the placental marginal hematomes. The submitted female dogs to this technique demonstrated normal puppy births without death.

Experimental infection of rabbits with a recombinant bovine herpesvirus type 5 (BoHV-5) gI, gE and US9-negative

Bovine herpesvirus type 5 (BoHV-5) is a major cause of viral meningoencephalitis in cattle. The expression of different viral proteins has been associated with BoHV-5 neuropathogenesis. Among these, gI, gE and US9 have been considered essential for the production of neurological disease in infected animals. To evaluate the role of gI, gE and US9 in neurovirulence, a recombinant from which the respective genes were deleted (BoHV-5 gI-/gE-/US9-) was constructed and inoculated in rabbits of two age groups (four and eight weeks-old). When the recombinant virus was inoculated through the paranasal sinuses of four weeks-old rabbits, neurological disease was observed and death was the outcome in 4 out of 13 (30.7 %) animals, whereas clinical signs and death were observed in 11/13 (84.6%) of rabbits infected with the parental virus. In eight weeks-old rabbits, the BoHV-5 gI-/gE-/US9- did not induce clinically apparent disease and could not be reactivated after dexamethasone administration, whereas wild type BoHV-5 caused disease in 55.5% of the animals and was reactivated. These findings reveal that the simultaneous deletion of gI, gE and US9 genes did reduce but did not completely abolish the neurovirulence of BoHV-5 in rabbits, indicating that other viral genes may also play a role in the induction of neurological disease.

Genital immunization of heifers with a glycoprotein Edeleted, recombinant bovine herpesvirus 1 strain confers protection upon challenge with a virulent isolate

Venereal infection of seronegative heifers and cows with bovine herpesvirus type 1.2 (BoHV-1.2) frequently results in vulvovaginitis and transient infertility. Parenteral immunization with inactivated or modified live BoHV-1 vaccines often fails in conferring protection upon genital challenge. We herein report an evaluation of the immune response and protection conferred by genital vaccination of heifers with a glycoprotein E-deleted recombinant virus (SV265gE-). A group of six seronegative heifers was vaccinated with SV265gE- (0,2mL containing 10(6.9)TCID50) in the vulva submucosa (group IV); four heifers were vaccinated intramuscularly (group IM, 1mL containing 10(7.6)TCID50) and four heifers remained as non-vaccinated controls. Heifers vaccinated IV developed mild, transient local edema and hyperemia and shed low amounts of virus for a few days after vaccination, yet a sentinel heifer maintained in close contact did not seroconvert. Attempts to reactivate the vaccine virus in two IV vaccinated heifers by intravenous administration of dexamethasone (0.5mg/kg) at day 70 pv failed since no virus shedding, recrudescence of genital signs or seroconversion were observed. At day 70 pv, all vaccinated and control heifers were challenged by genital inoculation of a highly virulent BoHV-1.2 isolate (SV56/90, 10(7.1)TCID50/animal). After challenge, virus shedding was detected in genital secretions of control animals for 8.2 days (8-9); in the IM group for 6.2 days (4-8 days) and during 5.2 days (5-6 days) in the IV group. Control non-vaccinated heifers developed moderate (2/4) or severe (2/4) vulvovaginitis lasting 9 to 13 days (x: 10.7 days). The disease was characterized by vulvar edema, vulvo-vestibular congestion, vesicles progressing to coalescence and erosions, fibrino-necrotic plaques and fibrinopurulent exudate. IM vaccinated heifers developed mild (1/3) or moderate (3/4) genital lesions, lasting 10 to 12 days (x: 10.7 days); and IV vaccinated heifers developed mild and transient vulvovaginitis (3/4) or mild to moderate genital lesions (1/4). In the IV group, the clinical signs lasted 4 to 8 days (x: 5.5 days). Clinical examination of the animals after challenge revealed that vaccination by both routes conferred some degree of protection, yet IV vaccination was clearly more effective in reducing the severity and duration of clinical disease. Furthermore, IV vaccination reduced the period of virus shedding in comparison with both groups. Taken together, these results demonstrate that SV265gE- is sufficiently attenuated upon IV vaccination in a low-titer dosis, is not readily reactivated after corticosteroid treatment and lastly, and more importantly, confers local protection upon challenge with a high titer of a virulent heterologous BoHV-1 isolate. Therefore, the use of this recombinant for genital immunization may be considered for prevention of BoHV-1-associated genital disease in the field.