RESUMEN
Managed populations of honey bees (Apis mellifera Linnaeus; Hymenoptera: Apidae) are regularly exposed to infectious diseases. Good hive management including the occasional application of antibiotics can help mitigate infectious outbreaks, but new beekeeping tools and techniques that bolster immunity and help control disease transmission are welcome. In this review, we focus on the applications of beneficial microbes for disease management as well as to support hive health and sustainability within the apicultural industry. We draw attention to the latest advances in probiotic approaches as well as the integration of fermented foods (such as water kefir) with disease-fighting properties that might ultimately be delivered to hives as an alternative or partial antidote to antibiotics. There is substantial evidence from in vitro laboratory studies that suggest beneficial microbes could be an effective method for improving disease resistance in honey bees. However, colony level evidence is lacking and there is urgent need for further validation via controlled field trials experimentally designed to test defined microbial compositions against specific diseases of interest.
Asunto(s)
Apicultura , Abejas , Fermentación , Microbioma Gastrointestinal , Probióticos , Animales , Antibacterianos/inmunología , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Apicultura/métodos , Abejas/efectos de los fármacos , Abejas/inmunología , Abejas/microbiología , Fermentación/inmunología , Microbioma Gastrointestinal/inmunología , Probióticos/farmacología , Probióticos/uso terapéuticoRESUMEN
Citrate is an ubiquitous compound in nature. However, citrate fermentation is present only in a few pathogenic or nonpathogenic microorganisms. The citrate fermentation pathway includes a citrate transporter, a citrate lyase complex, an oxaloacetate decarboxylase and a regulatory system. Enterococcus faecalis is commonly present in the gastro-intestinal microbiota of warm-blooded animals and insect guts. These bacteria can also cause infection and disease in immunocompromised individuals. In the present study, we performed whole genome analysis in Enterococcus strains finding that the complete citrate pathway is present in all of the E. faecalis strains isolated from such diverse habitats as animals, hospitals, water, milk, plants, insects, cheese, etc. These results indicate the importance of this metabolic preservation for persistence and growth of E. faecalis in different niches. We also analyzed the role of citrate metabolism in the E. faecalis pathogenicity. We found that an E. faecalis citrate fermentation-deficient strain was less pathogenic for Galleria mellonella larvae than the wild type. Furthermore, strains with deletions in the oxaloacetate decarboxylase subunits or in the α-acetolactate synthase resulted also less virulent than the wild type strain. We also observed that citrate promoters are induced in blood, urine and also in the hemolymph of G. mellonella. In addition, we showed that citrate fermentation allows E. faecalis to grow better in blood, urine and G. mellonella. The results presented here clearly indicate that citrate fermentation plays an important role in E. faecalis opportunistic pathogenic behavior.
Asunto(s)
Ácido Cítrico/metabolismo , Enterococcus faecalis/patogenicidad , Fermentación/genética , Infecciones por Bacterias Grampositivas/microbiología , Infecciones Oportunistas/microbiología , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Carboxiliasas/genética , Carboxiliasas/metabolismo , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Modelos Animales de Enfermedad , Enterococcus faecalis/genética , Enterococcus faecalis/inmunología , Enterococcus faecalis/metabolismo , Fermentación/inmunología , Regulación Bacteriana de la Expresión Génica , Genoma Bacteriano/genética , Infecciones por Bacterias Grampositivas/inmunología , Humanos , Redes y Vías Metabólicas/genética , Mariposas Nocturnas/inmunología , Mariposas Nocturnas/microbiología , Familia de Multigenes/genética , Infecciones Oportunistas/inmunología , Regiones Promotoras Genéticas/genética , Secuenciación Completa del GenomaRESUMEN
Giardiasis, caused by the protozoan Giardia intestinalis, is one of the most common intestinal diseases worldwide and constitutes an important problem for the public health systems of various countries. Kefir is a probiotic drink obtained by fermenting milk with 'kefir grains', which consist mainly of bacteria and yeasts that coexist in a complex symbiotic association. In this work, we studied the ability of kefir to protect mice from G. intestinalis infection, and characterized the host immune response to this probiotic in the context of the intestinal infection. Six- to 8-week-old C75BL/6 mice were separated into four groups: controls, kefir mice (receiving 1â:â100 dilution of kefir in drinking water for 14 days), Giardia mice (infected orally with 4×10(7) trophozoites of G. intestinalis at day 7) and Giardia-kefir mice (kefir-treated G. intestinalis-infected mice), and killed at 2 or 7 days post-infection. Kefir administration was able to significantly reduce the intensity of Giardia infection at 7 days post-infection. An increase in the percentage of CD4(+) T cells at 2 days post-infection was observed in the Peyer's patches (PP) of mice belonging to the Giardia group compared with the control and kefir groups, while the percentage of CD4(+) T cells in PP in the Giardia-kefir group was similar to that of controls. At 2 days post-infection, a reduction in the percentage of B220-positive major histocompatibility complex class II medium cells in PP was observed in infected mice compared with the other groups. At 7 days post-infection, Giardia-infected mice showed a reduction in RcFcε-positive cells compared with the control group, suggesting a downregulation of the inflammatory response. However, the percentages of RcFcε-positive cells did not differ from controls in the kefir and Giardia-kefir groups. An increase in IgA-positive cells was observed in the lamina propria of the kefir group compared with controls at 2 days post-infection. Interestingly, the diminished number of IgA-positive cells registered in the Giardia group at 7 days post-infection was restored by kefir feeding, although the increase in IgA-positive cells was no longer observed in the kefir group at that time. No significant differences in CXCL10 expression were registered between groups, in concordance with the absence of inflammation in small-intestinal tissue. Interestingly, a slight reduction in CCL20 expression was observed in the Giardia group, suggesting that G. intestinalis might downregulate its expression as a way of evading the inflammatory immune response. On the other hand, a trend towards an increase in TNF-α expression was observed in the kefir group, while the Giardia-kefir group showed a significant increase in TNF-α expression. Moreover, kefir-receiving mice (kefir and Giardia-kefir groups) showed an increase in the expression of IFN-γ, the most relevant Th1 cytokine, at 2 days post-infection. Our results demonstrate that feeding mice with kefir reduces G. intestinalis infection and promotes the activation of different mechanisms of humoral and cellular immunity that are downregulated by parasitic infection, thus contributing to protection.
Asunto(s)
Productos Lácteos Cultivados/inmunología , Fermentación/inmunología , Giardia lamblia/inmunología , Giardiasis/inmunología , Giardiasis/prevención & control , Leche/inmunología , Animales , Linfocitos B/inmunología , Linfocitos B/metabolismo , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Quimiocinas/inmunología , Quimiocinas/metabolismo , Productos Lácteos Cultivados/metabolismo , Regulación hacia Abajo/inmunología , Femenino , Genes MHC Clase II/inmunología , Giardia lamblia/metabolismo , Giardiasis/metabolismo , Inmunoglobulina A/inmunología , Inmunoglobulina A/metabolismo , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/prevención & control , Interferón gamma/inmunología , Interferón gamma/metabolismo , Intestino Delgado/inmunología , Intestino Delgado/metabolismo , Mastocitos/inmunología , Mastocitos/metabolismo , Ratones , Ratones Endogámicos C57BL , Leche/metabolismo , Membrana Mucosa/inmunología , Membrana Mucosa/metabolismo , Ganglios Linfáticos Agregados/inmunología , Ganglios Linfáticos Agregados/metabolismo , Factor de Necrosis Tumoral alfa/inmunología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
La levadura Candida guilliermondii es objeto de estudio debido a su capacidad de producir xilitol aprovechando compuestos hemicelulósicos ricos en xilosa, dado esto, la cepa Candida guilliermondii aislada del fruto del corozo chiquito (Bactris guineensis) fue usada en este estudio con el fin de evaluar su capacidad para producir xilitol sobre un sustrato hidrolizado de cascarilla de arroz. El objetivo de este trabajo fue determinar los parámetros fermentativos como producción de xilitol, productividad volumétrica (Qp) y rendimiento de sustrato en producto (Yp/s) durante la fermentación con la cepa nativa Candida guilliermondii. Se emplearon 200 ml de medio de cultivo hidrolizado de cascarilla de arroz, el cual contenía una concentración de xilosa de 27,5 g/L. La fermentación se llevó a cabo bajo las siguientes condiciones: temperatura 30 ºC, pH del medio 5,8, agitación 120 rpm e inóculo adaptado de 3 g/L. Los resultados mostraron que después de 120 horas de fermentación se obtuvieron 2,6 g/L de xilitol con productividad volumétrica (Qp) de 0,02 g/L-h y rendimiento de sustrato en producto (Yp/s) de 0,13 g/g. De esta manera, la cepa nativa Candida guilliermondii, aislada del fruto de Corozo chiquito (Bactris guineensis), produjo xilitol bajo condiciones específicas de fermentación.
The yeast Candida guilliermondii has been studied due to its ability to produce xylitol in xylose-rich hemicellulosic compounds, Candida guilliermondii strain isolated from the fruit of Corozo chiquito (Bactris guineensis) was used in this study to assess their ability to xylitol production on these substrates. The aim of this study was to determine the fermentation parameters such as xylitol production, volumetric productivity (Qp) and yield of xylitol production (Yp/s) during fermentation with the native strain Candida guilliermondii. Was used 200 ml of culture medium rice husk hydrolysate, which contained a xylose concentration of 27.5 g/L. The fermentation was carried out under the following conditions: temperature 30 ºC, pH of 5.8, agitation 120 rpm and adapted inoculum of 3 g/L. The results showed that after 120 hours of fermentation 2.6 g / L of xylitol was achieved with volumetric productivity (Qp) 0.02 g/L-h and 0.13 g/g yield of xylitol production (Yp/s). The native strain Candida guilliermondii, isolated from the fruit of Corozo chiquito (Bactris guineensis) produced xylitol fermentation under specific conditions.
Asunto(s)
Fermentación/fisiología , Fermentación/genética , Fermentación/inmunología , Xilosa/análisis , Xilosa/análogos & derivados , Xilosa/clasificación , Xilosa/fisiología , Levadura Seca/análisis , Levadura Seca/clasificación , Levadura Seca/farmacología , Levadura Seca/genética , Levadura Seca/provisión & distribución , Levadura Seca/química , Levadura Seca/síntesis químicaRESUMEN
En el presente trabajo se describe la producción de las enzimas fitasa, celulasa, xilanasa y proteasa con Aspergillus ficuum cepa DSM 932 mediante fermentación en estado sólido (SSF) usando torta de canola y pomaza de cranberry como sustratos. Como medida indirecta de la producción de las enzimas se usó en cada caso la actividad enzimática. la torta de canola resultó ser un mejor sustrato para fitasa, celulasa y xilanasa, en tanto que la pomaza de cranberry resultó ser un sustrato potencial para proteasa. Mediante ultrafiltración escalonada fue posible purificar parcialmente los extractos enzimáticos de fitasa, celulasas y xilanasas, obtenidos a partir de torta de canola. La fitasa resultó tener un tamaño >100 kDa, en tanto que las celulasas y xilanasas presentan actividad en los retenidos de 10, 30 y 50 kDa, lo que indicaría que las isoenzimas de ambos complejos tienen pesos moleculares que oscilan entre 10 y 100 kDa.
In this paper, describes the production of the enzymes phytase, cellulase, xylanase and protease by Aspergillus ficuum DSM 932 strain, in solid state fermentation (SSF) using canola cake and cranberry pomace as substrates. The enzyme activity was used in each case as an indirect measure of the enzymes production. Canola meal turned out to be a better substrate for phytase, cellulase and xylanase, while cranberry pomace was found to be a potential substrate for protease. Various ultrafiltration operations were carried out, decreasing the cut off membranes out in order to purify partially extracts of enzymes phytase, cellulase and xylanase, obtained from canola meal. Phytase was found to have a size >100 kDa, whereas cellulase and xylanase activity present in the retained 10, 30 and 50 kDa, suggesting that isozymes of both complexes have molecular weights ranging between 10 and 100 kDa.
Asunto(s)
/análisis , Agroindustria/análisis , Agroindustria/efectos adversos , Agroindustria/métodos , Celulasa/análisis , /análisis , Fermentación/genética , Fermentación/inmunologíaRESUMEN
Se describe la producción de fitasa mediante cultivos del tipo sumergido (SmF) y sobre sustrato sólido (SSF) con Aspergillus ficuum DSM 932 en medios de cultivos basados en residuos de la agroindustria. La actividad enzimática fitásica se usó como medida indirecta de la producción de la enzima. En SmF, pH 5,3 y 25 ºC, se trabajó en fermentadores de diferentes volúmenes y con el mayor se operó con diferentes niveles de aireación y agitación. En SSF a 25 ºC se usaron placas de Petri. En SmF con un medio basado en cereales se presentó la mejor actividad neta (0,25 FTU/mL) al sexto día para 300 rpm y 0,5 vvm. En SSF, la torta de canola resultó ser el mejor sustrato con una actividad fitásica neta máxima al tercer día de 6,79 FTU/mL de extracto, equivalente a 33,96 FTU/g de sustrato sólido o 56,43 FTU/g de sustrato seco. Aplicando tecnologías de membrana se concentró un extracto de fitasa a partir de una SmF en medio basado en cereales y también fue posible purificar 6,33 veces un extracto de fitasa producido en SSF con torta de canola, diafiltrando tres veces consecutivas el retenido de 100 kDa. La enzima fitasa de la cepa A. ficuum DSM 932 mostró tener un tamaño ⥠100 kDa.
Phytase production by submerged fermentation (SmF) and solid state fermentation (SSF) using Aspergillus ficuum DSM 932 in agro-waste-based culture media is described here. Phytase enzyme activity was used for the indirect measurement of enzyme production. Fermentation was carried out in SmF, pH 5.3 at 25 ºC with two fermenters having different volumes; the largest one had different levels of aeration and agitation. Petri dishes were used for SSF at 25 °C. A cereal-based medium obtained the best net activity (0.25 FTU mL-1) for SmF on the sixth day at 300 rpm at 0.5 vvm. Canola cake was the best substrate for SSF, having maximum net phytase activity on the third day: 6.79 FTU mL-1 extract, equivalent to 33.96 FTU g-1 solid substrate or 56.43 FTU g-1 dry substrate. A phytase extract was concentrated from an SmF-based medium in cereals by applying membrane technologies. A phytase extract produced in SSF with canola cakes was purified 6.33 times using three consecutive diafiltrations of the 100 kDa retentate. A. ficuum DSM 932 phytase was ⥠100 kDa in size.
Asunto(s)
Fermentación/fisiología , Fermentación/genética , Fermentación/inmunologíaRESUMEN
BACKGROUND: Fermented milks containing probiotic bacteria are a way of delivering bioactive constituents to targets in the gastrointestinal tract. We reported previously that the fermentation of milk at constant pH 6 by L. helveticus R389 increased its content of peptide fractions, and the oral administration of the non-bacterial fraction (FMSpH6) to mice increased total secretory IgA in the intestinal lumen and enhanced the number of IgA and various cytokines producing cells as well as the secretion of IL-6 by small intestine epithelial cells. We also demonstrated that this FMSpH6 was effective for the prevention of Salmonella typhimurium infection in mice. In this work, we studied in mice the impact of the oral administration of the supernatant of milk fermented by L. helveticus R389 on the gut physiology by measuring parameters such as calcium channels and E-cadherin expression, the activation of the biological signal calcineurin and mast and goblet cells, as a way to determine some mechanisms involved in the immunomodulating effects of the milk fermentation products, observed in previous studies. We analyzed the impact of the supernatant of milk fermented by L. helveticus R389 at pH6-controlled on the expression of calcineurin and on the reinforcement of the ephitelial barrier, measuring parameters such as calcium channels and E-cadherin expression and in the reinforcement of the non-specific immunity determining mast cells and goblet cells associated to the gut. RESULTS: We observed an enhanced expression of TRPV6 channels in the duodenum, indicating an improved capacity for dietary Ca2+ uptake. We demonstrated an enhanced expression of calcineurin in the small intestine, able to upregulate immune parameters such as IL-2 and TNF production, with an increase in the number of these cytokines secreting cells. We determined an increase in the number of mucosal mast cells and goblet cells, which would mean an improved state of mucosal surveillance at sites of infection. CONCLUSION: The oral administration of the supernatant of milk fermented by L. helveticus R389 enhanced the gut mucosal immunity by improving the mechanisms that reinforce the epithelial and non-specific barriers and the gut functioning at sites of infection, with an improvement in the expression of the enzyme calcineurin, an important signal in the network that activates the gut immune system. The results of this work contribute to revealing the mechanisms underlying the immunomodulation of the gut immune function by fermented milks with probiotic bacteria.
Asunto(s)
Calcineurina/metabolismo , Productos Lácteos Cultivados/inmunología , Fermentación/inmunología , Inmunidad Mucosa , Mucosa Intestinal/inmunología , Lactobacillus helveticus/fisiología , Animales , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD8-positivos/metabolismo , Cadherinas/metabolismo , Productos Lácteos Cultivados/química , Productos Lácteos Cultivados/microbiología , Técnica del Anticuerpo Fluorescente , Células Caliciformes/metabolismo , Interleucina-2/metabolismo , Mastocitos/inmunología , Mastocitos/metabolismo , Ratones , Ratones Endogámicos BALB C , Probióticos , Canales Catiónicos TRPV/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Sugarcane bagasse was used as substrate for xylanase production by means of a strain of Trichoderma harzianum Rifai isolated from decaying Aspidosperma sp. (peroba) wood. The bagasse was washed, dried, milled and wetted with minimal salts medium and the cultures grown at 28 ñ 2§C for 7 days. Two extraction methods were tested for enzyme recovery: (A) Tween 80, 0.1(per cent) (v/v), in physiological saline, and (B) 50mM sodium acetate buffer, pH 5.0, under agitation (180rpm) for 15, 30 and 60min. After a single extraction, both extraction methods recovered an average of 15U/ml of xylanase activity, independent on the time of shaking. A second and third extraction recovered 10.4 and 6.6U/ml xylanase, respectively. The effect of volume size for extraction, and sugarcane bagasse concentration, on xylanase production were also investigated. The growth profile of Trichoderma harzianum was followed over 20 days on 14(per cent) (w/v) bagasse, and highest xylanase activity (288U/ml) appeared on the seventh day. The enzymatic extract after precipitation with ammonium sulphate was submitted to electrophoresis on polyacrylamide gels and showed 4 protein-staining bands, one of which exhibited xylanase activity.
Asunto(s)
Enzimas , Técnicas In Vitro , Industria del Azúcar , Trichoderma , Activación Enzimática/inmunología , Fermentación/inmunologíaRESUMEN
Os alimentos funcionais constitutem hoje prioridade de pesquisa em todo mundo com a finalidade de elucidar as propriedades e os efeitos que estes produtos podem apresentar na promoçäo da saúde. As bactérias probióticas säo microrganismos vivos que, quando consumidos, exercem efeitos benéficos sobre o hospedeiro conferindo propriedades à microbiota endógena. Algumas propriedades benéficas atribuídas às culturas probióticas necessitam de estudos mais controlados para serem definitivamente esclarecidas. Neste artigo säo enfocados os aspectos tecnológicos dos probióticos, os efeitos associados ao consumo de produtos contendo probióticos e as principais cepas empregadas. Säo apresentadados resultados experimentais...
Asunto(s)
Microbiología de Alimentos , Alimentos Fortificados , Técnicas In Vitro , Lactobacillus , Leche , Probióticos/análisis , Probióticos/metabolismo , Análisis de los Alimentos/métodos , Medios de Cultivo , Estudio de Evaluación , Fermentación/inmunologíaRESUMEN
Níveis intracelulares de G-3-PDH (sn-glicerol-3-fosfato: NAD+ 2-oxidoredutase, EC 1.1.1.8) de levedura de panificaçäo foram acompanhados durante a estocagem sob três diferentes temperaturas. Semelhantes valores de biomassa final e de atividade específica da enzima foram obtidos após crescimento por 48 horas de duas linhagens de leveduras de panificaçäo. O melhor meio (meio indutor) para obtençäo de G-3-PDH foi: extrato de levedura (1 por cento, p/v), peptona (2 por cento, p/v), glicerol (3 por cento, v/v) e etanol (1 por cento v/v). O choque osmótico com adiçäo de NaCl 0,6 M provocou aumento da atividade de G-3-PDH de 1,2 vezes para leveduras crescidas em meio indutor por 48 horas e transferidas...
Asunto(s)
Industria de Alimentos , Glicerolfosfato Deshidrogenasa , Almacenamiento de Materiales y Suministros , Pan/microbiología , Saccharomyces cerevisiae , Biomasa , Técnicas de Cultivo de Célula , Fermentación/inmunología , Liofilización , Manejo de EspecímenesRESUMEN
Se midió la degradación in vitro de enzimas fibrolíticas exógenas y su efecto en la degradación in vitro de FDN y FDA de heno de alfalfa o ballico. Se usó la primera fase de Tilley y Terry (0,3,6,12,24,48 y 72h) con saliva McDougall (S) sola o con líquido ruminal (LR). La desaparición de la enzima (E) fue constante de 0 a 6h, y aumentó de 12 a 72h. La concentración de N-NH4 fue constante de 0 a 24h y su mayor valor fue a las 72h. La desaparición de FDN de los forrajes se incrementó de 6 a 72h con la E y de 24 a 72h con E + LR. Además, E aumentó la desaparición de FDA de alfalfa de 3 a 72h y la del ballico de 3 a 12h; pero E + LR no cambio la desaparición de FDA. La E con LR aumentó la desaparición neta de FDN de ambos forrajes a las 48 y 72h, pero redujo la desaparición neta de FDA del ballico a las 12h, en tanto que no hubo diferencias para la FDA de la alfalfa. En las primeras 12h no se dirigieron las enzimas del producto enzimático, el cual tiene un efecto positivo importante en la digestibilidad in vitro de la pared celular del heno de alfalfa y de ballico, aún en presencia de microorganismos ruminales