Níveis crescentes de parede de levedura sobre a resposta imune celular e perfil hematológico de frangos de corte / Cellular immune response and hematological parameters of broilers from different age broiler breeders, fed with a sorghum meal with different yeast wall levels

Um experimento foi conduzido para avaliar o efeito de níveis crescentes de parede de levedura e idade das matrizes reprodutoras sobre o peso dos órgãos linfóides, a resposta imune celular e o perfil hematológico de frangos de corte. Foram utilizados 3.360 pintos de corte da linhagem Cobb, distribuídos em delineamento inteiramente casualisado, em esquema fatorial 2x5, mais dois controles, sendo duas idades de matrizes (34 e 57 semanas de idade) e cinco níveis de suplementação de parede de levedura (zero, um, dois, três e quatro kg de parede de levedura/tonelada de ração). A idade das matrizes influenciou a resposta de todas as variáveis. A inclusão de 3kg de parede de levedura/tonelada de ração promoveu, na progênie de reprodutoras de 57 semanas, reação inflamatória mais intensa quando comparada a dieta controle, no entanto não houve aumento significativo no número de heterófilos e linfócitos circulantes. Conclui-se que a utilização da parede de levedura associada ao sorgo ou não em rações de frangos de corte ainda necessita de estudos complementares, que incluam, por exemplo, os componentes purificados da parede de levedura (MOS e ß-glucano).

An experiment was carried out to evaluate the effect increasing levels of yeast wall and broiler breeders' age levels on lymphoid organs weight, cellular immune response and hematological parameters in broilers. A total of 3,360 Cobb broilers were allotted, in a completely randomized design and a 2x5 factorial arrangement, and two controls, compound of two broiler breeders age (34 and 57 weeks of age) and five yeast wall levels (zero, one, two, three and four kg of yeast wall/ton of diet). Broiler breeders' age affected all studied variables. The inclusion of 3kg of yeast wall/ton of meal increased, at 57 weeks age broiler breeders, more intense inflammatory reaction when compared to control diet; however the circulated heterophils and lymphocytes numbers were not increased. In conclusion, the use of yeast wall, associated or not to sorghum on broilers diet still needs complementary studies, for example, purified components of yeast wall (MOS and ß-glucano).

Contribuição da citologia de papanicolaou para o diagnóstico de leveduras em secreção vaginal / Contribution of the cytology of papanicolaou for the diagnosis of yeasts in vaginal secretion

Introdução: candidíase vulvovaginal (CVV) é uma infecção causada pelo crescimento anormal de fungos do tipo leveduras na mucosa do trato genital. Ultimamente, tem crescido o interesse na utilização da citologia de Papanicolaou (Pap) no diagnóstico de infecções associadas a patógenos de transmissão sexual, mas o método tem sido pouco avaliado para CVV. Objetivos: avaliar a acurácia diagnóstica da citologia de Papanicolaou para o diagnóstico de fungos vaginais, bem como comparar os resultados da citologia com os da bacterioscopia de secreção vaginal (Gram) e cultura. Métodos: foram coletadas secreções cérvico-vaginais para Gram, Papanicolaou e cultura de leveduras, e as colônias crescidas na cultura foram contadas. As mulheres não foram triadas para sintomatologia de CVV e foram descartadas aquelas com outra infecção agente ou imunodeficiências. Foram realizados cálculos estatísticos de sensibilidade, especificidade, valores preditivo positivo e negativo, e freqüência de falso-positivos e negativos, em três situações: na detecção de fungos independente da quantidade de colônias; nos casos com crescimento de 1-9 colônias; e naqueles com crescimento de ? 100. Resultados: a cultura foi positiva em 35 pacientes (21,70%). Sem considerar o número de colônias, a sensibilidade do Gram e da citologia foram muito parecidas (67,65% e 62,85%), nos casos com crescimento de 1-9 colônias, de 37,50% e 50,00%, e nos casos com crescimento ? 100 colônias, de86,61% e 78,26%. Conclusão: a citologia mostrou-se um método morfológico com eficiência muito semelhante e para pequenos números de colônias melhor do que o Gram para detectar leveduras vaginais, tornando-a perfeitamente aplicável na rotina laboratorial para este diagnóstico.

Introduction: vulvovaginal Candidiasis vulvovaginal (VVC) it is an infection caused by the abnormal growth of fungus of the type yeasts in themucous membrane of the genital treatment. Lately, it has been increasing the interest in the use of the cytology of Papanicolaou (Pap) in the diagnosis of associated infections the pathogens of sexual transmission, but the method has been little appraised for VVC. Objectives: to evaluate the accuracy diagnostic of the cytology of Papanicolaou for the diagnosis of vaginal fungus, as well as to compare the results of the cytology with the one of the bacterioscopy of vaginal secretion (Gram) and culture. Methods: cervico-vaginal secretions were collected for Gram, Papanicolaou and culture of yeasts, and the grown colonies in the culture were counted. The women didn?t go selected to simptomatology of VVC and they were discarded those with other infection agent or immunodeficiency. Statistical calculations of sensibility were accomplished, specificity, values positive and negative predictive, and frequency of false positive and negative, in three situations: in the independent detection of fungus of the amount of colonies; in thecases with growth of 1-9 colonies; and in those with growth of ? 100. Results: the culture of it was positive in 35 patients (21,70%). Without considering the number of colonies, the sensibility of Gram and of the cytology they were very similar (67,65% and 62,85%), in the cases with growth of 1-9 colonies, of 37,50% and 50,00%, and in the cases with growth ? 100 colonies, of 86,61% and 78,26%. Conclusion: the cytology was shown a morphologic method with very similar efficiency and for small numbers of colonies better than Gram to detect vaginal yeasts, turning her perfectly applicable in the routine laboratorial for this diagnosis.

Cell differentiation increases astrocyte phagocytic activity: a quantitative analysis of both GFAP labeling and pas-stained yeast cells

Since efficiency of phagocytic potential in activated astrocytes is still a subject of controversy, an attempt was made to quantify simultaneously phagocytic activity and astrocyte differentiation. Resorting to Junin virus, known to induce astrocyte activation, infected vs control samples of cultured rat astroglial cells were serially harvested up to day 12 post-inoculation (pi), and subjected to a triple staining procedure consisting in immunoperoxidase labeling of GFAP, periodic acid-Schiff (PAS) reaction in added baker's yeast cells and hematoxylin for nuclear staining of the whole cell monolayer. Adopting GFAP labeling as a specific marker of astrocyte differentiation, the immunoprecipitate development over time was measured. Direct calculation of the initial reaction rate was feasible given its linear behavior during the first 10 min, so that GFAP amount was regarded proportional to peroxidase activity. As determined by digital image analysis, mean optical density (MOD) values of GFAP in infected samples increased from 0.618 + 0.082 at day 1 pi to 0.825 + 0.125 at day 3, leveling off at 1.010 + 0.101 as from day 9, while control unifected samples remained unchanged at roughly 0.6 during the entire observation period. In turn, phagocytosis was quantified by PAS staing densitometry, whose intensity varied according to wall degradation of yeast cells. MOD levels of PAS-stained phagocytized yeast cells were significantly lower (p <0.05) in efected vs control cultures at 48 and 72 h following their addition to the astroglial monolayer. According to simultaneous quantification of two components of astrocyte response to viral infection, it is concluded that phagocytic activity increases with astrocyte differentiation.

Flow cytometry analysis of cell cycle in myosin II-deficient yeast

OBJECTIVE: To determine whether cell cycle changes can be detected in myosin II-deficient cells using flow cytometry techniques. BACKGROUND: Although the primary role of myosin II (Myo1p) in the yeast Saccharomyces cerevisiae is in cytokinesis we have reported that this conventional myosin also appears to inuence the regulation of cell wall metabolism as indicated by increases in the expression of chitin metabolizing enzymes in a null mutant of the MYO1 gene. The expression of these enzymes is known to be regulated in the cell cycle suggesting that cell cycle changes may alter their expression. METHODS: Flow cytometry was employed to assess the nuclear DNA content of logarithmic yeast cell cultures as a means of determining changes in the cell cycle of Myo1p-deficient cells. RESULTS: Significant changes were observed in the Myo1p-deficient strain suggesting that these cells are arrested in G2/M-phase of the cell cycle. CONCLUSIONS: Based on the results of this preliminary study, we propose a model in which the increased activity of chitin metabolizing enzymes may be explained by a mitotic arrest in these cells.

Assesment of viability of yeast by flow cytometry using light green as vital dye exclusion

Se compararon dos técnicas para valorar la viabilidad de levaduras por medio de citometría de flujo aplicada a microbiología. En una se usó el colorante yoduro de propidio (YP), el cual al haber un daño de membrana pasa al citoplasma para unirse al DNA o RNA, lo que permite excluir microorganismos dañados de los íntegros, y en la otra se usó el colorante verde claro (VC), ampliamente utilizado en tinciones histológicas, histoquímicas e inmunoquímicas. El modelo de levadura utilizado fue el de muerte por calor en Candida guilliermondii. El análisis de citometría de flujo por diagramas de puntos, en ambos colorantes mostró una separación muy clara de las poblaciónes muertas hacia verde-naranja (350-600 nm) para el VC y rojo (550-725 nm) para el YP, en los canales de fluorescencia correspondientes. Todos los experimentos mostraron entre las poblaciones de levadura vivas y muertas por calor, una prueba de Kolmogorov-Smirnov (K-S) mayor de 99 por ciento de probabilidad de diferencia entre los histogramas de los organismos vivos de los muertos, con una D= con valores de 0.50-0.64 para VC y de 0.91-0.98 para YP. Dado el costo y riesgo de carcinogénesis por el YP, el VC puede ser utilizado en el análisis de viabilidad en levaduras por citofluorometría

Una técnica para preparaciones permanentes de levaduras / A technique for permanent slides of yeasts

Modificando una técnica indicada por Arta-gaveitía-Allende para microcultivos de levaduras, se obtuvieron entre lamina y laminilla, preparaciones teñidas permanentes que mantienen su integridad por largo tiempo