Influencia de las concentraciones de melaza de Saccharum officinarum L. en la producción de Blastoesporas de Isaria fumorosea en un biorreactor
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Date
2024
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Universidad Nacional de Trujillo
Abstract
Se evaluó la influencia de las concentraciones de melaza al 1%, 5% y 10% de Saccharum
officinarum L. en la producción de blastosporas de Isaria fumosorosea en un biorreactor tipo
tanque agitado y aireado de 30 Lt. Para esto, se aisló Isaria fumosorosea a partir de cadáveres
momificados de larvas del insecto plaga Spodoptera frugiperda en hojas de uva (Vitis vinifera
L.). Luego se realizaron subcultivos de Isaria fumosorosea en placas petri conteniendo Agar Papa
Dextrosa para la obtención de esporas las cuales fueron cosechadas agregando 10 mL de Tween 80
estéril (0,05%) sobre la superficie de agar y con un asa de drigalski estéril se raspó
suavemente y la suspensión se recolectó en frascos estériles de 250 mL. Luego, la suspensión
se ajustó a una concentración de 5 × 108 conidios/ml con agua destilada estéril usando una
cámara de neubauer. Para la producción de blastosporas en medio líquido se usó 3
tratamientos el cual consiste en distintas concentraciones de melaza (1%, 5% y 10%). Cada
medio de cultivo fue esterilizado in situ a 121°C, 15 min, 15 psi en un biorreactor de 30 Lt.
Finalizado el proceso de esterilización, se dejó enfriar el medio de cultivo hasta que alcanzó
una temperatura entre 28-30 °C. Posteriormente, se inoculó con 1 lt de la suspensión de
esporas de Isaria fumosorosea a una concentración de 5 × 108 conidios/ml. Se dejó incubar
durante 3 días a 25°C a 350 rpm y pH 5,3. Los cultivos fueron evaluados a las 24, 48 y 72
h para la producción de blastosporas. Todos los experimentos se repitieron tres veces de
forma independiente (n=3) por cada tratamiento. Los resultados indican que Isaria
fumosorosea a la concentración del 10% de melaza favorece la producción de blastosporas
(3,59x109 blastosporas/mL) a las 72 h de incubación por lo que podría ser usado para la
formulación de micoinsecticidas de uso agrícola.
The influence of molasses concentrations at 1%, 5%, and 10% of Saccharum officinarum L. on the production of blastospores of Isaria fumosorosea in a 30 L stirred and aerated tank bioreactor was evaluated. For this, Isaria fumosorosea was isolated from cadavers of the pest insect larvae Spodoptera frugiperda on grapevine (Vitis vinifera L.) leaves. Subcultures of Isaria fumosorosea were then performed on petri dishes containing Potato Dextrose Agar to obtain spores, which were harvested by adding 10 mL of sterile Tween 80 (0.05%) on the agar surface. The surface was gently scraped with a sterile Drigalski spatula, and the suspension was collected in sterile 250 mL flasks. The suspension was then adjusted to a concentration of 5 × 10⁸ conidia/mL with sterile distilled water using a Neubauer chamber. For the production of blastospores in liquid medium, three treatments were used, consisting of different molasses concentrations (1%, 5%, and 10%). Each culture medium was sterilized in situ at 121°C for 15 minutes at 15 psi in a 30 L bioreactor. After the sterilization process, the culture medium was allowed to cool until it reached a temperature between 28-30°C. Subsequently, it was inoculated with 1 L of the Isaria fumosorosea spore suspension at a concentration of 5 × 10⁸ conidia/mL. It was incubated for 3 days at 25°C, 350 rpm, and pH 5.3. The cultures were evaluated at 24, 48, and 72 hours for blastospore production. All experiments were independently repeated three times (n=3) for each treatment. The results indicate that Isaria fumosorosea at a 10% molasses concentration favors blastospore production (3.59 × 10⁹ blastospores/mL) after 72 hours of incubation, which could be used for the formulation of agricultural mycoinsecticides.
The influence of molasses concentrations at 1%, 5%, and 10% of Saccharum officinarum L. on the production of blastospores of Isaria fumosorosea in a 30 L stirred and aerated tank bioreactor was evaluated. For this, Isaria fumosorosea was isolated from cadavers of the pest insect larvae Spodoptera frugiperda on grapevine (Vitis vinifera L.) leaves. Subcultures of Isaria fumosorosea were then performed on petri dishes containing Potato Dextrose Agar to obtain spores, which were harvested by adding 10 mL of sterile Tween 80 (0.05%) on the agar surface. The surface was gently scraped with a sterile Drigalski spatula, and the suspension was collected in sterile 250 mL flasks. The suspension was then adjusted to a concentration of 5 × 10⁸ conidia/mL with sterile distilled water using a Neubauer chamber. For the production of blastospores in liquid medium, three treatments were used, consisting of different molasses concentrations (1%, 5%, and 10%). Each culture medium was sterilized in situ at 121°C for 15 minutes at 15 psi in a 30 L bioreactor. After the sterilization process, the culture medium was allowed to cool until it reached a temperature between 28-30°C. Subsequently, it was inoculated with 1 L of the Isaria fumosorosea spore suspension at a concentration of 5 × 10⁸ conidia/mL. It was incubated for 3 days at 25°C, 350 rpm, and pH 5.3. The cultures were evaluated at 24, 48, and 72 hours for blastospore production. All experiments were independently repeated three times (n=3) for each treatment. The results indicate that Isaria fumosorosea at a 10% molasses concentration favors blastospore production (3.59 × 10⁹ blastospores/mL) after 72 hours of incubation, which could be used for the formulation of agricultural mycoinsecticides.
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