RESUMO
Background: The development of animal reproductive biotechniques is intended of raising reproduction effi ciency. The mammal ovary contains thousands of follicles, of which approximately 99.9% are eliminated by means of the atresia, apoptosis and cellular necrosis process. In other to reduce this follicular loss, the methods for isolation and characterization of preantral follicles have been studied as a premise to culture systems of these structures. The purpose of this study was quantify and evaluate the quality of preantral ovarian follicles from prepubertal gilts after mechanical isolation procedure. Furthermore, it aims to analyze the preantral follicular histological morphology.Materials, Methods & Results: Ovaries (n = 20) from prepubertal gilts were divided in two halves and used each one for isolation and histological processes. The tissue chopper previously regulated for the performance of seriated cuts at 200 m intervals was used for mechanical isolation. The marker bisbenzimidine Hoechst 33342 was added to the follicular pool for evidence the presence of granulosa cells around oocyte and the viability of the isolated PAF was evaluated by using propidium iodide. For the histological evaluation, the ovarian halves were fi xed, dehydrated and diaphanized, and after enclosed in paraffi n, each of them was divided into 5 blocks and sectioned in series 6 m thick. At
Background: The development of animal reproductive biotechniques is intended of raising reproduction effi ciency. The mammal ovary contains thousands of follicles, of which approximately 99.9% are eliminated by means of the atresia, apoptosis and cellular necrosis process. In other to reduce this follicular loss, the methods for isolation and characterization of preantral follicles have been studied as a premise to culture systems of these structures. The purpose of this study was quantify and evaluate the quality of preantral ovarian follicles from prepubertal gilts after mechanical isolation procedure. Furthermore, it aims to analyze the preantral follicular histological morphology.Materials, Methods & Results: Ovaries (n = 20) from prepubertal gilts were divided in two halves and used each one for isolation and histological processes. The tissue chopper previously regulated for the performance of seriated cuts at 200 m intervals was used for mechanical isolation. The marker bisbenzimidine Hoechst 33342 was added to the follicular pool for evidence the presence of granulosa cells around oocyte and the viability of the isolated PAF was evaluated by using propidium iodide. For the histological evaluation, the ovarian halves were fi xed, dehydrated and diaphanized, and after enclosed in paraffi n, each of them was divided into 5 blocks and sectioned in series 6 m thick. At
RESUMO
Background: The development of animal reproductive biotechniques is intended of raising reproduction effi ciency. The mammal ovary contains thousands of follicles, of which approximately 99.9% are eliminated by means of the atresia, apoptosis and cellular necrosis process. In other to reduce this follicular loss, the methods for isolation and characterization of preantral follicles have been studied as a premise to culture systems of these structures. The purpose of this study was quantify and evaluate the quality of preantral ovarian follicles from prepubertal gilts after mechanical isolation procedure. Furthermore, it aims to analyze the preantral follicular histological morphology.Materials, Methods & Results: Ovaries (n = 20) from prepubertal gilts were divided in two halves and used each one for isolation and histological processes. The tissue chopper previously regulated for the performance of seriated cuts at 200 m intervals was used for mechanical isolation. The marker bisbenzimidine Hoechst 33342 was added to the follicular pool for evidence the presence of granulosa cells around oocyte and the viability of the isolated PAF was evaluated by using propidium iodide. For the histological evaluation, the ovarian halves were fi xed, dehydrated and diaphanized, and after enclosed in paraffi n, each of them was divided into 5 blocks and sectioned in series 6 m thick. At
Background: The development of animal reproductive biotechniques is intended of raising reproduction effi ciency. The mammal ovary contains thousands of follicles, of which approximately 99.9% are eliminated by means of the atresia, apoptosis and cellular necrosis process. In other to reduce this follicular loss, the methods for isolation and characterization of preantral follicles have been studied as a premise to culture systems of these structures. The purpose of this study was quantify and evaluate the quality of preantral ovarian follicles from prepubertal gilts after mechanical isolation procedure. Furthermore, it aims to analyze the preantral follicular histological morphology.Materials, Methods & Results: Ovaries (n = 20) from prepubertal gilts were divided in two halves and used each one for isolation and histological processes. The tissue chopper previously regulated for the performance of seriated cuts at 200 m intervals was used for mechanical isolation. The marker bisbenzimidine Hoechst 33342 was added to the follicular pool for evidence the presence of granulosa cells around oocyte and the viability of the isolated PAF was evaluated by using propidium iodide. For the histological evaluation, the ovarian halves were fi xed, dehydrated and diaphanized, and after enclosed in paraffi n, each of them was divided into 5 blocks and sectioned in series 6 m thick. At
RESUMO
The aim of present work was to evaluate the changes of cervico-vaginal epithelial cells and progesterone levels of nuliparous goats submitted to artificial photoperiod, in Middle-West of Brazil. Seventh Alpine and six crossbreed nuliparous goats were used, distributed in two groups: control (GC), maintained at isolated stall and submitted to natural light for the time of the year, and treated (GT), submitted to an alternate regime among natural and artificial light, for 24 hours and 35 uninterrupted days. Cérvico-vaginal content was collected, prepared on glass slides and differential cellular counts were carried out on Papanicolau smears. The progesterone concentration in blood serum was measurement by ELISA. Parabasal were the most frequent cell type present, followed by intermediate cells, mainly in the end of metaestrus and diestrus. Superficial cornified cells were present during proestrus and estrus. The cellular profile and progesterone concentrations oscillated in agreement with estrus cycle phase in control animals, but confuse on the begin of experimental period for treated animals. Cytological evaluation of cervico-vaginal content could be of value for a hormonal bioassay of estrus cycle of goats submitted to artificial photoperiod just after the adaptation period. KEY WORDS: Artificial photoperiod, caprine, hormone, vaginal cytology.
O objetivo do presente trabalho foi avaliar as alterações celulares do epitélio cérvico-vaginal e as concentrações de progesterona sérica em cabras nulíparas expostas ao fotoperíodo artificial, na região Centro-Oeste do Brasil. Dezessete cabras da raça Alpina e seis mestiças, nulíparas, foram aleatoriamente distribuídas em grupo-controle (GC), mantido em galpão isolado e submetido ao regime de luz natural para a época do ano, e grupo tratado (GT), submetido ao regime alternado entre luz natural e artificial, por 24 horas durante 35 dias ininterruptos. Realizou-se colheita de conteúdo cérvico-vaginal para preparo de esfregaços, que foram corados pelo método Papanicolau. A concentração sérica de progesterona foi mensurada por enzimaimunoensaio. Verificou-se predomínio de células parabasais, seguido de células intermediárias, principalmente no final do metaestro e diestro. Células superficiais queratinizadas foram relacionadas às fases de proestro e estro. O perfil celular acompanhou as oscilações das concentrações séricas de progesterona, as quais sofreram modificações de acordo com a fase do ciclo estral nos animais-controle, mas com um período confuso no início da exposição à luz artificial, nos animais tratados. O estudo das modificações celulares do epitélio cérvico-vaginal pode ser valioso como um bioensaio hormonal do ciclo estral em cabras submetidas ao fotoperíodo artific
RESUMO
The aim of present work was to evaluate the changes of cervico-vaginal epithelial cells and progesterone levels of nuliparous goats submitted to artificial photoperiod, in Middle-West of Brazil. Seventh Alpine and six crossbreed nuliparous goats were used, distributed in two groups: control (GC), maintained at isolated stall and submitted to natural light for the time of the year, and treated (GT), submitted to an alternate regime among natural and artificial light, for 24 hours and 35 uninterrupted days. Cérvico-vaginal content was collected, prepared on glass slides and differential cellular counts were carried out on Papanicolau smears. The progesterone concentration in blood serum was measurement by ELISA. Parabasal were the most frequent cell type present, followed by intermediate cells, mainly in the end of metaestrus and diestrus. Superficial cornified cells were present during proestrus and estrus. The cellular profile and progesterone concentrations oscillated in agreement with estrus cycle phase in control animals, but confuse on the begin of experimental period for treated animals. Cytological evaluation of cervico-vaginal content could be of value for a hormonal bioassay of estrus cycle of goats submitted to artificial photoperiod just after the adaptation period. KEY WORDS: Artificial photoperiod, caprine, hormone, vaginal cytology.
O objetivo do presente trabalho foi avaliar as alterações celulares do epitélio cérvico-vaginal e as concentrações de progesterona sérica em cabras nulíparas expostas ao fotoperíodo artificial, na região Centro-Oeste do Brasil. Dezessete cabras da raça Alpina e seis mestiças, nulíparas, foram aleatoriamente distribuídas em grupo-controle (GC), mantido em galpão isolado e submetido ao regime de luz natural para a época do ano, e grupo tratado (GT), submetido ao regime alternado entre luz natural e artificial, por 24 horas durante 35 dias ininterruptos. Realizou-se colheita de conteúdo cérvico-vaginal para preparo de esfregaços, que foram corados pelo método Papanicolau. A concentração sérica de progesterona foi mensurada por enzimaimunoensaio. Verificou-se predomínio de células parabasais, seguido de células intermediárias, principalmente no final do metaestro e diestro. Células superficiais queratinizadas foram relacionadas às fases de proestro e estro. O perfil celular acompanhou as oscilações das concentrações séricas de progesterona, as quais sofreram modificações de acordo com a fase do ciclo estral nos animais-controle, mas com um período confuso no início da exposição à luz artificial, nos animais tratados. O estudo das modificações celulares do epitélio cérvico-vaginal pode ser valioso como um bioensaio hormonal do ciclo estral em cabras submetidas ao fotoperíodo artific
RESUMO
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.
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.
RESUMO
Este estudo foi desenvolvido com o objetivo de verificar as alterações presentes na microbiota cérvico-vaginal durante o final da gestação e o puerpério em vacas da raça Girolando. Para isso, no período compreendido entre27 dias antes do parto e 58 dias após, foram realizados 117 cultivos bacteriológicos, oriundos de material colhido da região cérvico-vaginal a intervalos de 14 dias, com exceção do período compreendido entre o dia zero (parto) e o dia 2. Foram obtidos 101 isolamentos, constatando-se que as amostras negativas predominaram durante o período pré-parto (58,8%) e no final do puerpério (44%), mas que durante o período entre o parto e o 16o dia todas as amostras colhidas foram positivas ao cultivo e isolamento microbiano.Os agentes isolados foram: Escherichia coli (27,72%),Staphylococcus coagulase negativa (21,78%),Staphylococcus coagulase positiva (1,98%), Streptococcus spp. (19,80%), Corynebacterium spp. (7,92%), Corynebacterium bovis (3,98%), Bacillus spp. (6,98%),Pseudomonas spp. (3,96%), Enterobacter aerogenes (1,98%), Actinomyces pyogenes (1,98%), Citrobacter freundii (0,99%) e Citrobacter amalonaticus (0,99%). Amostras obtidas de fêmeas primíparas mostraram maior positividade, sugerindo sensibilidade alterada dessa categoria à contaminação do trato reprodutor por microrganismos no período subseqüente ao parto. PALAVRAS-CHAVE: Bovinos, microbiota cérvico-vaginal
RESUMO
Este estudo foi desenvolvido com o objetivo de verificar as alterações presentes na microbiota cérvico-vaginal durante o final da gestação e o puerpério em vacas da raça Girolando. Para isso, no período compreendido entre27 dias antes do parto e 58 dias após, foram realizados 117 cultivos bacteriológicos, oriundos de material colhido da região cérvico-vaginal a intervalos de 14 dias, com exceção do período compreendido entre o dia zero (parto) e o dia 2. Foram obtidos 101 isolamentos, constatando-se que as amostras negativas predominaram durante o período pré-parto (58,8%) e no final do puerpério (44%), mas que durante o período entre o parto e o 16o dia todas as amostras colhidas foram positivas ao cultivo e isolamento microbiano.Os agentes isolados foram: Escherichia coli (27,72%),Staphylococcus coagulase negativa (21,78%),Staphylococcus coagulase positiva (1,98%), Streptococcus spp. (19,80%), Corynebacterium spp. (7,92%), Corynebacterium bovis (3,98%), Bacillus spp. (6,98%),Pseudomonas spp. (3,96%), Enterobacter aerogenes (1,98%), Actinomyces pyogenes (1,98%), Citrobacter freundii (0,99%) e Citrobacter amalonaticus (0,99%). Amostras obtidas de fêmeas primíparas mostraram maior positividade, sugerindo sensibilidade alterada dessa categoria à contaminação do trato reprodutor por microrganismos no período subseqüente ao parto. PALAVRAS-CHAVE: Bovinos, microbiota cérvico-vaginal
RESUMO
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.
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.
RESUMO
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.
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.
RESUMO
Testosterone concentration was measured by radioimmunoassay in 30 samples of blood serum obtained five times at each 6 hours interval during 24 hours from 6 Jaffarabadi x Mediterranean buffalo bulls. Blood samplings were carried out twice, respectively, in one day of summer and one day of winter season. Testosterone concentration ranged from 0.10 to 1.36 ng/ml in the winter day and from 0.10 to 2.54 ng/ml in the summer day. Maximum values were obtained at 6:00 p.m. (0.52) in summer and 0.82 ng/ml in winter, then abrupt drops occurred, first at 12 m. (0.37) in summer and 0.21 in winter, and the second one at 24 p.m. (0.17) in summer and 0.49 ng/ml in winter. In summer, higher serum testosterone levels were observed during the day.
Concentrações de testosterona foram determinadas por radioimunoensaio em 30 amostras de soro sangüíneo obtidas cinco vezes durante 24 horas, de 6 búfalos adultos Jaffarabadi x Mediterrâneo. As amostras foram obtidas durante um dia do inverno e um dia do verão de 1991. As concentrações de testosterona variaram de 0,10 a 1,36 ng/ml no inverno e de 0,10 a 2,454 ng/ml no verão. Valores máximos foram obtidos às 6,00 horas (0,52 ng/ml) no verão e 0,82 ng/ml no inverno, ocorrendo então duas quedas abruptas, a primeira às 12 horas (0,37 ng/ml) no verão e 0,21 ng/ml no inverno e a segunda 24 horas (0,17 ng/ml) no verão e 0,49 ng/ml no inverno. No verão, níveis mais altos de testosterona sérica foram observados durante o dia.