Histopatology of the reproductive tract of Nellore pubertal heifers with genital ureaplasmosis

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.

Assessment of cow and farm level risk factors associated with Ureaplasma diversum in pasture-based dairy systems - A field study

ABSTRACT Potential risk factors for Ureaplasma diversum in the vaginal mucus of 1,238 dairy cows were included in a multivariate logistic regression model, based on the cow level (i.e., granular vulvovaginitis [+GVV], yearly milk production [4500 kg or more], pregnancy, predominance of Bos taurus [+Bos Taurus], score of corporal condition [at least 2.5], concomitant positivity for Escherichia coli [+E.coli]), and farm level i.e., milking room hygiene (-Milking room), dunghill location, and replacement female). Ureaplasma diversum was present in 41.1% of the samples. Independent risk factors for U. diversum were +GVV (odds ratio [OR], 1.31); +Mycoplasma spp (OR, 5.67); yearly milk production (4500 kg or more) (OR, 1.99); +Bos taurus (OR, 1.68); +E. coli (OR, 4.96); -milking room (OR, 2.31); and replacement females (OR, 1.89). Ureaplasma diversum vaginal colonization was strongly associated with Mycoplasma spp., E. coli, and number of pregnant cows.

Embryonic stem-like cells derived from in vitro produced bovine blastocysts

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 percent) 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.

Construção de iniciadores e otimização de ensaios de PCR e de nested-PCR para a detecção específica de Tritrichomonas foetus / Primers design and optimization of PCR and nested-PCR assays for the specific detection of Tritrichomonas foetus

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.

Fertilidade real e intervalo de partos de vacas nelore PO sob manejo extensivo e sem estação de monta na região centro oeste do Brasil / True fertility and calving interval of nelore po cows under extensive management and without breeding season in middle west of Brazil

Este estudo foi desenvolvido com o objetivo de avaliar as fontes de variação que interferem no índice fertilidade real (FR) e o efeito do ambiente no intervalo de partos (IDP) de vacas Nelore PO criadas em sistema extensivo de produção na região Centro-Oeste do Brasil. Os dados analisados foram ordem de parto (OP), idade ao parto(IP), peso ao nascimento do bezerro (PN), peso à desmama (PD), ano e mês do parto (AP, MP) e sexo do bezerro (SB) de546 vacas, calculando-se o IDP e fixando-se o efeito aleatório da mãe (EAM). IDP e PD foram utilizados para calcular o índice de fertilidade real. O IDP médio de 452,68 ± 117,10 dias foi influenciado (P<0,05) pela ordem de parto, idade aoparto, peso ao nascer e efeito aleatório da mãe. A FR média de 148,6 ± 34,5kg sofreu influência (P<0,05) de ano e mês do parto, ordem de parto, sexo do bezerro e idade da mãe.

This study was designed to evaluate the variation sources that interfere in the true fertility indexand the effect of herd environment in the calving interval of Nelore P.O. cows raised in extensive production systems inMiddle-West of Brazil, without breeding season. The analyzed data were parity order (OP), calving age (IP), calf birthweight (PN), weaning weight (PD), calving year and month (AP, MP) and calf sex (SB) of 546 cows, calculating the calving interval (IDP) and fixing the aleatory effect of mother (EAM). IDP and PD were used to calculate the index oftrue fertility (FR). IDP (452,68±117,10 days) was influenced (P <0,05) by OP, IP, PN and EAM. FR average(148,6±34,5kg) was influenced (P <0,05) by AP, MP, OP, S and IM.

Estudo experimental do enxerto autólogo de ovário em cadelas submetidas a ovariectomia e ovariosalpingohisterectomia / Experimental ovarian autograft in bitches submitted to ovariectomy and ovariohysterectomy

A pesquisa mostra os resultados obtidos em 10 cadelas mestiças das quais, 6 foram castradas e enxertadas com fragmento autólogo de ovário e 4 formaram os grupos de controle, somente castradas. Foram avaliadas no período pós-operatório de 12 meses através exames clínico e citológico, dosagem sérica de progesterona, observaçäo macroscópica do local de enxertia e exame histológico de fragmento do enxerto ovariano. Durante a fase pós-operatória de observaçäo, as fêmeas somente castradas e as enxertadas nao mostraram alteraçoes das mamas, com relaçäo ao peso comporal, utilizando o teste de Wilcoxon as somente castradas demonstraram diferença estatísticamente significante (p=0,0156). No tocante a citologia vaginal as fêmeas enxertadas apresentaram as quatro fases do ciclo estral. Através o teste de Wilcoxon concluiu-se que ocorre diferença significativa nas dosagens séricas de progesterona entre as fêmeas somente castradas (p=0.0082) e as enxertadas (p=0.0156). Após período de 12 e 36 meses foi realizada avaliaçäo macroscópica e exame histológico dos enxertos ovarianos, que mostraram diferentes etapas de foliculogênese.