ABSTRACT In order to study and characterize the lesions in the reproductive tract of Nellore heifers naturally infected with Ureaplasma diversum and presenting granular vulvovaginitis syndrome (GVS), fragments of uterine tube, uterus, cervix, vagina and vulva of 20 animals were evaluated. The macroscopic lesions of the vulvovaginal mucosa were classified in scores of “1” mild, until “4”, severe inflammation and pustular or necrotic lesions. The histopathological evaluation was performed using scores of “1” to “4”, according to the inflammatory alterations. The fragments with severe microscopic lesions (3 and 4) were from the uterine tubes and uterus, which showed leukocytes infiltration and destruction and/or necrosis of epithelium. Alterations in the lower reproductive tract fragments were mild, but characteristics of acute inflammatory processes. The histopathological findings of the reproductive tract of females naturally infected with Ureaplasma diversum are consistent with injuries that compromise the environment from the local where spermatozoa acquires ability to fertilize an oocyte until those where the oocyte is fertilized. Therefore, animals with GVS should be identified early in the herd, because, besides the reduction in the fertility rates caused by tissue damages, they can contribute to disseminate the microorganism. Key words: bovine, tissue evaluation, reproduction, Ureaplasma diversum.

The aim of this work was to study the derivation of bovine embryonic stem-like (ES-like) cells from the inner cell mass (ICM) of in vitro produced blastocysts. The ICMs were mechanically isolated and six out of seventeen (35%) ICMs could attach to a monolayer of murine embryonic fibroblasts (MEF). Ten days after, primary outgrowths were mechanically dissected into several small clumps and transferred to a new MEF layer. Cells were further propagated and passaged by physical dissociation over a 60 days period. The pluripotency of the bovine ES-like cells was confirmed by RT-PCR of Oct-4 and STAT-3 gene markers. The colonies were weakly stained for alkaline phosphatase and the mesoderm and endoderm differentiation gene markers such as GATA-4 and Flk-1, respectively, were not expressed. Embryoid bodies were spontaneously formed at the seventh passage. Results showed that bovine ES-like cells could be obtained and passaged by mechanical procedures from the fresh in vitro produced blastocysts.

Tritrichomonas foetus é um protozoário patogênico responsável por doença venérea em bovinos conhecida por tricomonose genital bovina. A tricomonose bovina é uma doença venérea causada pelo protozoário cujo habitat natural é o trato genital. Os protocolos já desenvolvidos para o diagnóstico deste parasito por PCR, apesar de serem eficazes na identificação do DNA genômico alvo, promovem algumas amplificações inespecíficas ou são incapazes de distinguir T. foetus das outras espécies do gênero. O presente trabalho foi desenvolvido com o objetivo de estabelecer e otimizar protocolos de ensaio de PCR e nested-PCR para o diagnóstico específico de T. foetus, empregando-se novos iniciadores, selecionados do alinhamento das seqüências dos genes 18S rRNA, 5,8S rRNA, 28S rRNA e dos espaços transcritos do rDNA (ITS1 e ITS2). Um par de iniciadores foi construído para amplificação gênero-específica de um fragmento de 648 pares de base e outros dois para a obtenção de produtos espécie- específicos de 343 e 429 pb. Nenhuma reação cruzada foi observada frente ao DNA genômico de Bos taurus ou de microrganismos responsáveis por infecções genitais. A sensibilidade dos ensaios de PCR e de nested-PCR apresentados neste estudo permitiu um limiar de detecção de até dois parasitos.

Tritrichomonas foetus is a pathogenic protozoan that causes a venereal disease in cattle known as bovine genital tricomonosis. In spite of the efficacy to recognize the target genomic DNA, the protocols so far developed for the diagnosis of this organism by PCR promote some inespecific amplifications or they are unable to discriminate T. foetus against other species within the genus. The objective of this study was to assess and optimize PCR and nested-PCR assays for the specific diagnosis of T. foetus, using novel primers selected from the alignment of sequences of the genes 18S rRNA, 5.8S rRNA, 28S rRNA and of the internal transcribed spacers of the rDNA (ITS1 and ITS2). A pair of primers was constructed for the genus-specific amplification of a 648 bp fragment and two others to amplify T. foetus species-specific fragments of 343 and 429 bp. No cross amplification was observed against Bos taurus genomic DNA neither against the DNA of usual bovine genital pathogens. Both, single and nested-PCR assays, presented analytical sensitivity to detect at least two T. foetus organisms.

O presente estudo objetivou descrever as alterações físico-químicas da urina, identificando os agentes etiológicos presentes na urina de matrizes descartadas sem causa definida. De 35 amostras examinadas, 16 (45,7%) foram positivas para presença de sangue e/ou nitrito (positivo para infecção urinária, IU); 8 (50%) tiveram coloração amarela escura e 6 (37,5%) coloração amarela clara; 2 (12,5%) amostras não apresentavam alteração. Dez amostras tinham odor amoniacal, das quais 7 foram positivas para IU. Também houve prevalência de urinas turvas e turvas com grumos, bem como de pH normal (5,5 - 6,5) e densidades maiores que 1,012. Presença de microrganismos foi verificada em 91,5% das 35 amostras examinadas, sendo a Escherichia coli (45,7%), a bactéria com maior prevalência. Em 11 amostras (31,4%), verificou-se a presença de Actinobaculum suis; destas, 4 foram simultaneamente positivas para IU e Actinobaculum suis; 7 foram negativas para IU, observando-se porém, a presença de Actinobaculum suis pelo exame de imunofluorescência indireta.

The present study describes the physical-chemical alterations of the urine and identifies etiological agents of the urine of 35 sows. Out of 35 samples, 16 (45.7%) were positive for nitrite and/or blood, positive for urinary infection (UI); eight (50%) were dark yellow and six (37.5%) had clear yellow coloration; in two (12.5%) samples did not show any alteration. Ten samples had ammonia smell, seven among them were positive for UI. Other findings were turbid urine and turbid with grumes, normal pH values (5.5 - 6.5) and densities higher than 1012. Microorganisms were isolated in 91.5% out of 35 urine samples, with prevalence of Escherichia coli (45.7%). In 11 samples (31.4%) Actinobaculum suis was isolated, four were simultaneously positive for UI and Actinobaculum suis; seven were negative for UI, but it was observed the presence of Actinobaculum suis in the indirect immunofluorescence.