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United States Department of Agriculture

Agricultural Research Service

Research Project: PRODUCTION, STABILIZATION AND FORMULATION OF MICROBIAL AGENTS AND THEIR NATURAL PRODUCTS

Location: Crop Bioprotection Research

Title: Dissolved oxygen levels affect dimorphic growth by the entomopathogenic fungus Isaria fumosorosea

Author
item JACKSON, MARK

Submitted to: Biocontrol Science and Technology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: November 16, 2011
Publication Date: January 10, 2012
Repository URL: http://handle.nal.usda.gov/10113/55137
Citation: Jackson, M.A. 2012. Dissolved oxygen levels affect dimorphic growth by the entomopathogenic fungus Isaria fumosorosea. Biocontrol Science and Technology. 22(1):67-79. DOI: 10.1080/09583157.2011.642339.

Interpretive Summary: The fungus Isaria fumosorosea (Ifr) infects and kills numerous insect pests including whiteflies, aphids, Asian citrus psyllids, and subterranean termites. For development as a biological control agent for these insects, a low-cost production method for Ifr is required. Using liquid fermentation, Ifr grows as a mass of filaments or as individual cells called blastospores. Blastospores are the desired form of Ifr when used in spray applications to control insects. In this study, we evaluated the effect of aeration on the production of the blastospores of Ifr. High aeration rates supported Ifr growth in the blastospore form during liquid fermentation with very little filamentous growth. High concentrations of stable blastospores were produced in short fermentation times using low-cost nutrients when high aeration rates were used during the fermentation process. These studies showed that adequate aeration is critical for the production of blastospores of Ifr. Better blastospore yields using low cost nutrients will significantly lower production costs for Ifr blastospores and improve their potential for commercial use against important insect pests. Lower cost, natural, non-chemical controls for these serious insect pests will provide farmers, greenhouse operators, and homeowners with additional management tools.

Technical Abstract: The entomopathogenic fungus Isaria fumosorosea is capable of dimorphic growth (hyphal or yeast-like) in submerged culture. In shake flask studies, we evaluated the impact of aeration on the mode of growth of I. fumosorosea. Using 250 mL baffled Erlenmeyer flasks, culture volumes of 50, 100, 150, and 200 mL of I. fumosorosea were grown in a shaker incubator at 350 rpm and 28 °C. Dissolved oxygen (DO) was continuously monitored using a non-invasive oxygen monitoring system. Culture volumes of 50 mL maintained DO concentrations above 10% throughout the 3 day growth period. Fifty mL cultures accumulated biomass and produced blastospores more rapidly than the other culture volumes tested and more than 55% of their biomass was blastospores. Dissolved oxygen was depleted in culture volumes of 100, 150, and 200 mL after a growth period of 20.5, 16.8 and 13.5 hours, respectively. In 100 mL cultures, DO was below 10% for 8 hours whereas DO in 150 and 200 mL cultures were exhausted (< 3%) for the remainder of culture growth resulting in significantly lower blastospore yields and increased hyphal growth. These studies were used to set oxygen concentration requirements (> 20% DO) for I. fumosorosea growth in a 100 L bioreactor resulting in comparable blastospore production with minimal hyphal growth. Maintaining higher DO levels resulted in increased blastospore production by cultures of I. fumosorosea grown on nitrogen sources (cottonseed meal and soy flour) that previously elicited excessive hyphal growth. These studies have shown that oxygen availability is essential for significant yeast-like growth by I. fumosorosea cultures and that continuous monitoring of oxygen concentrations in shake flask cultures can be a useful tool for establishing appropriate aeration conditions for use in bioreactors.

Last Modified: 9/10/2014
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