Effects of Carboxymethylcellulose and Carboxypolymethylene on Morphology of <Emphasis Type="Italic">Aspergillus fumigatus</Emphasis> NRRL 2346 and Fumagillin Production

Aspergillus fumigatus NRRL 2346 is the producer of fumagillin, an antitumor antibiotic that inhibits angiogenesis. This strain is very difficult to grow reproducibly in shake flasks owing to an extreme form of pellet growth and extensive wall growth. The effects of carboxymethylcellulose (CMC) and carboxypolymethylene (Carbopol) on growth and fumagillin production by A. fumigatus were investigated. By adding the polymers to the fermentation medium, the growth form of the mold was changed from a single large glob to small reproducible pellets, and wall growth was diminished to a minimum. Carbopol, at a lower concentration, was more effective than CMC in improving both morphology and production. Small pellets were produced which favored fumagillin biosynthesis. 1.5% (wt/vol) CMC and 0.3% (wt/vol) Carbopol were found to be the optimum concentrations; higher levels increased viscosity to an unacceptable level. Received: 1 November 2001 / Accepted: 27 March 2002     

Impact of Carboxymethylcellulose on Morphology and Antibiotic Production by Streptomyces hygroscopicus

A chemically defined media consisting of carboxymethylcellulose (CMC) was developed to maximize the production of antibiotics, hexaene H-85 and azalomycine, by Streptomyces hygroscopicus CH-7. The production of antibiotics by filamentous organisms is often dependent on the morphology and size distribution of the pellet population within the culture. By adding the polymer to the fermentation medium, the growth was changed from a single large glob to small reproducible pellets, and wall growth was diminished to a minimum. Maximum concentrations of hexaene H-85 (146.7 mg/dm(3)) and azalomycine (188.6 mg/dm(3)) were reached at 3.0% and 1.0% (w/v) CMC, respectively.     

Carbon and nitrogen source nutrition of fumagillin biosynthesis by Aspergillus fumigatus

The carbon and nitrogen source requirements of Aspergillus fumigatus NRRL 2436 for growth and production of the angiogenesis inhibitor fumagillin were studied in chemically defined media. Both carbon and nitrogen sources strongly influenced fumagillin formation. Two out of 29 carbon sources tested interfered with fumagillin biosynthesis. The best combination of two carbon sources was 30 g L(-1) xylan and 50 g L(-1) mannose. Of fifteen nitrogen sources tested, three ammonium salts (chloride, sulfate, and dibasic phosphate) failed to support fumagillin formation, presumably due to the low pH which developed. The dosage-response study of the best nitrogen source, L-glutamic acid, revealed that 9 g L(-1) was optimal. Volumetric production of fumagillin was increased by 15-fold over that in the starting (Peterson-Goldstein) medium as a result of these findings.     

Induction and Regeneration of Autoplasts from Clostridium thermohydrosulfuricum JW102 and Thermoanaerobacter ethanolicus JW200

Autolysis was induced to form stable, cell wall-free cells of Clostridium thermohydrosulfuricum JW102 and Thermoanaerobacter ethanolicus JW200, using a complex medium containing glycine (0.4% wt/vol) and/or sucrose or glycerol (10% wt/vol) at an optimum temperature of 64°C. Autoplasts of both bacteria were grown as L-phase colonies on solid medium; more than 50% of these colonies regenerated to the walled form during prolonged incubation. The removal of the cell wall was confirmed by electron microscopy.     

Effects of polymer additives on fermentation parameters in a culture of A. niger

Of 24 different polymer and surfactant materials examined, a carboxypolymethylene (“Carbopol”) was found to cause enhancement of respiration rates in an Aspergillus niger culture by as much as 200%. Enhancement of other fermentation parameters, such as cellular growth and amylase production, was also observed. The enhancement effects of Carbopol were examined with clusters of spores and mold pellets. In the first case, it appears that the ionized carboxyl groups of Carbopol induced electrostatic repulsion among the spores thus initiating pulp growth with increased interfacial area of contact between the mold and the nutrient medium. In the second case, the Carbopol additive formed a thin film attached to the surface of the pellets which seemed to be responsible for an increased rate of potassium transport and, hence, fermentation yields. Additive utilization as substrate and physiological changes in the culture were not observed in these cases. It was also found that the probability of pellet formation, the size of pellets formed, and the number of spores per pellet can be correlated to the energy input to the fermentation system.     

Improving dispersed growth of Frankia using Carbopol

In laboratory culture, strains of the actinomycete Frankia grow slowly and form dense mycelial pellets. In this study, we have shown that by adding the anionic polymer Carbopol 941® to liquid growth media, dispersed growth of Frankia is improved. Cell protein measurements indicated higher biomass production in cultures maintained in media supplemented with Carbopol. Fluorescence staining and microscopy were used to compare viability of well-dispersed mycelia and pellets.     

Molecular Cloning and Expression of Cellulase Genes from Ruminococcus albus 8 in Escherichia coli Bacteriophage lambda

A genomic library of Ruminococcus albus 8 DNA was constructed by using the Escherichia coli bacteriophage lambdaDASH. Recombinants were screened for cellulolytic activity by plating in soft agar (0.7%) overlays containing either 1% (wt/vol) carboxymethyl cellulose (CMC), 4-methylumbelliferyl-beta-d-cellobioside (MUC, 1 mg/ml), or 1% (wt/vol) Ostazin brilliant red-hydroxyethyl cellulose (OBR-HEC). One hundred and three recombinant phage exhibiting activity against OBR-HEC were found, and these fell into different classes based on the size of the zone of hydrolysis. Twenty-one recombinant phage exhibiting activity against CMC and 19 recombinant phage exhibiting activity against MUC were isolated. Four OBR-HEC, five CMC, and seven MUC clones were further analyzed by restriction endonuclease mapping and cellulase substrate specificity to identify unique clones and to determine their cellulase type. Three different clone types representing endoglucanase activity were identified. Three clones that appeared to encode exoglucanase type activity and four clones that had a mixed specificity, including beta-glucosidase activity, were also identified.     

Effects of different spices used in production of fermented sausages on growth of and curvacin A production by Lactobacillus curvatus LTH 1174

Lactobacillus curvatus LTH 1174, a fermented sausage isolate, produces the listericidal bacteriocin curvacin A. The effect of different spices relevant for the production of fermented sausages was investigated in vitro through laboratory fermentations with a meat simulation medium and an imposed pH profile relevant for Belgian-type fermented sausages. The influence on the growth characteristics and especially on the kinetics of curvacin A production with L. curvatus LTH 1174 was evaluated. Pepper, nutmeg, rosemary, mace, and garlic all decreased the maximum specific growth rate, while paprika was the only spice that increased it. The effect on the lag phase was minor except for nutmeg and especially for garlic, which increased it, yet garlic was stimulatory for biomass production. The maximum attainable biomass concentration (X(max)) was severely decreased by the addition of 0.40% (wt/vol) nutmeg, while 0.35% (wt/vol) garlic or 0.80% (wt/vol) white pepper increased X(max). Nutmeg decreased both growth and bacteriocin production considerably. Garlic was the only spice enhancing specific bacteriocin production, resulting in higher bacteriocin activity in the cell-free culture supernatant. Finally, lactic acid production was stimulated by the addition of pepper, and this was not due to the manganese present because an amount of manganese that was not growth limiting was added to the growth medium. Addition of spices to the sausage mixture is clearly a factor that will influence the effectiveness of bacteriocinogenic starter cultures in fermented-sausage manufacturing.     

Optimization of cholesterol removal by probiotics in the presence of prebiotics by using a response surface method

Lactobacillus casei ASCC 292 was grown in the presence of six prebiotics, namely, sorbitol, mannitol, maltodextrin, high-amylose maize, fructooligosaccharide (FOS), and inulin, in order to determine the combination of probiotic and prebiotics that would remove the highest level of cholesterol. A first-order model showed that the combination of L. casei ASCC 292, FOS, and maltodextrin was the most efficient for the removal of cholesterol, and the optimum experimental region was developed by using the steepest ascent. This led to the middle points of probiotic (1.70% [wt/vol]), FOS (4.80% [wt/vol]), and maltodextrin (6.80% [wt/vol]) for the development of a central composite design for optimization. Perturbation plot, response surface, and coefficient estimates showed that all three factors had significant quadratic effects on cholesterol removal, with FOS showing the most conspicuous quadratic change. A second-order polynomial regression model estimated that the optimum condition of the factors for cholesterol removal by L. casei ASCC 292 is 1.71% (wt/vol) probiotic, 4.95% (wt/vol) FOS, and 6.62% (wt/vol) maltodextrin. Validation experiments showed that the predicted optimum conditions were more efficient than the high and low levels of the factors and the center points. A response surface method proved reliable for developing the model, optimizing factors, and analyzing interaction effects. Analyses of growth, substrate utilization, growth yield, mean doubling time, and short-chain fatty acid (SCFA) production by the use of quadratic models indicated that cholesterol removal was growth associated. The concentration of L. casei ASCC 292 had the most significant quadratic effect on all responses studied, except for substrate utilization and SCFA production, which were significantly (P < 0.05) influenced by the interactions between the probiotic and both prebiotics, indicating that they were closely associated with the uptake of prebiotics.     

Inhibition of Aflatoxin Production by Surfactants

The effect of 12 surfactants on aflatoxin production, growth, and conidial germination by the fungus Aspergillus flavus is reported. Five nonionic surfactants, Triton X-100, Tergitol NP-7, Tergitol NP-10, polyoxyethylene (POE) 10 lauryl ether, and Latron AG-98, reduced aflatoxin production by 96 to 99% at 1% (wt/vol). Colony growth was restricted by the five nonionic surfactants at this concentration. Aflatoxin production was inhibited 31 to 53% by lower concentrations of Triton X-100 (0.001 to 0.0001%) at which colony growth was not affected. Triton X-301, a POE-derived anionic surfactant, had an effect on colony growth and aflatoxin production similar to that of the five POE-derived nonionic surfactants. Sodium dodecyl sulfate (SDS), an anionic surfactant, and dodecyltrimethylammonium bromide, a cationic surfactant, suppressed conidial germination at 1% (wt/vol). SDS had no effect on aflatoxin production or colony growth at 0.001%. The degree of aflatoxin inhibition by a surfactant appears to be a function of the length of the hydrophobic and hydrophilic chains of POE-derived surfactants.     

Effects of Different Spices Used in Production of Fermented Sausages on Growth of and Curvacin A Production by Lactobacillus curvatus LTH 1174

Lactobacillus curvatus LTH 1174, a fermented sausage isolate, produces the listericidal bacteriocin curvacin A. The effect of different spices relevant for the production of fermented sausages was investigated in vitro through laboratory fermentations with a meat simulation medium and an imposed pH profile relevant for Belgian-type fermented sausages. The influence on the growth characteristics and especially on the kinetics of curvacin A production with L. curvatus LTH 1174 was evaluated. Pepper, nutmeg, rosemary, mace, and garlic all decreased the maximum specific growth rate, while paprika was the only spice that increased it. The effect on the lag phase was minor except for nutmeg and especially for garlic, which increased it, yet garlic was stimulatory for biomass production. The maximum attainable biomass concentration (X_max) was severely decreased by the addition of 0.40% (wt/vol) nutmeg, while 0.35% (wt/vol) garlic or 0.80% (wt/vol) white pepper increased X_max. Nutmeg decreased both growth and bacteriocin production considerably. Garlic was the only spice enhancing specific bacteriocin production, resulting in higher bacteriocin activity in the cell-free culture supernatant. Finally, lactic acid production was stimulated by the addition of pepper, and this was not due to the manganese present because an amount of manganese that was not growth limiting was added to the growth medium. Addition of spices to the sausage mixture is clearly a factor that will influence the effectiveness of bacteriocinogenic starter cultures in fermented-sausage manufacturing.     

Optimization of Cholesterol Removal by Probiotics in the Presence of Prebiotics by Using a Response Surface Method

Lactobacillus casei ASCC 292 was grown in the presence of six prebiotics, namely, sorbitol, mannitol, maltodextrin, high-amylose maize, fructooligosaccharide (FOS), and inulin, in order to determine the combination of probiotic and prebiotics that would remove the highest level of cholesterol. A first-order model showed that the combination of L. casei ASCC 292, FOS, and maltodextrin was the most efficient for the removal of cholesterol, and the optimum experimental region was developed by using the steepest ascent. This led to the middle points of probiotic (1.70% [wt/vol]), FOS (4.80% [wt/vol]), and maltodextrin (6.80% [wt/vol]) for the development of a central composite design for optimization. Perturbation plot, response surface, and coefficient estimates showed that all three factors had significant quadratic effects on cholesterol removal, with FOS showing the most conspicuous quadratic change. A second-order polynomial regression model estimated that the optimum condition of the factors for cholesterol removal by L. casei ASCC 292 is 1.71% (wt/vol) probiotic, 4.95% (wt/vol) FOS, and 6.62% (wt/vol) maltodextrin. Validation experiments showed that the predicted optimum conditions were more efficient than the high and low levels of the factors and the center points. A response surface method proved reliable for developing the model, optimizing factors, and analyzing interaction effects. Analyses of growth, substrate utilization, growth yield, mean doubling time, and short-chain fatty acid (SCFA) production by the use of quadratic models indicated that cholesterol removal was growth associated. The concentration of L. casei ASCC 292 had the most significant quadratic effect on all responses studied, except for substrate utilization and SCFA production, which were significantly (P < 0.05) influenced by the interactions between the probiotic and both prebiotics, indicating that they were closely associated with the uptake of prebiotics.     

Investigations on neomycin production with immobilized cells of<Emphasis Type="Italic">Streptomyces marinensis</Emphasis> Nuv-5 in calcium alginate matrix

The purpose of this investigation was to study the effect ofStreptomyces marinensis NUV-5 cells immobilized in calcium alginate for the production of neomycin. The effect of various parameters, such as the effect of alginate concentration (1%, 2%, 3%, 4%, and 5% wt/vol), the effect of cation (caCl₂, BaCl₂, and SrCl₂), the concentration of cation (0.01M, 0.125M, 0.25M, 0.375M, and 0.5M), the curing times (1, 6, 11, 16, and 21 hours), and the diameter of the bead (1.48, 2.16, 3.24, 4.46, and 5.44 mm), on neomycin production and bead stability were studied. The effect of maltose (4%, 3%, 2%, and 1% wt/vol) and sodium glutamate (0.6%, 0.3%, 0.15%, and 0.075%) wt/vol) concentration on neomycin production was also studied. Better neomycin production was achieved with optimized parameters, such as alginate at 2% wt/vol, 0.25M CaCl₂, 1-hour curing time, and 3.24 mm bead diameter. Effective neomycin production was achieved with 3% wt/vol maltose and 0.6% wt/vol sodium glutamate concentration. The repeated batch fermentations were conducted (every 96 hours) using the optimized alginate beads, employing the production medium with 3% wt/vol maltose and 0.6% wt/vol sodium glutamate along with minerals salts solution. The increase in antibiotic production was observed up to the 5th cycle, and later gradual decrease in antibiotic production was observed. Comparison of the total antibiotic production with free cells and immobilized cells was also done. An enhanced antibiotic productivity of 32% was achieved with immobilized cells over the conventional free-cell fermentation, while 108% more productivity was achieved over the washed free-cell fermentation. From these results it is concluded that the immobilized cells ofS marinensis NUV-5 in calcium alginate are more efficient for the production of neomycin with repeated batch fermentation.     

Potent anti-angiogenic action of AGM-1470: comparison to the fumagillin parent.

The anti-angiogenic activity of AGM-1470, a new synthetic analog of fumagillin isolated from Aspergillus fumigatus, was extensively examined both in vitro and in vivo using four different types of assay and compared to that of the fumagillin parent. Locally administered AGM-1470 inhibited the angiogenesis in the chick embryo chorioallantoic membrane assay and the rat corneal assay. In the rat sponge implantation assay, systemically administered AGM-1470 inhibited angiogenesis induced by basic fibroblast growth factor. Furthermore, in the rat blood vessel organ culture assay, AGM-1470 (1-1,000 ng/ml) was found to selectively inhibit the capillary-like tube formation of endothelial cells with a minimal effect on the non-endothelial cell growth. AGM-1470 showed more potent anti-angiogenic activity and less toxicity than the fumagillin parent. Therefore, AGM-1470 is much better than the fumagillin parent as anti-angiogenic compound.     

Effects of CMC addition on bacterial cellulose production in a biofilm reactor and its paper sheets analysis

Bacterial cellulose (BC) can be grown into any desired shape such as pellicles, pellets, and spherelike balls, depending on the cultivation method, additives, and cell population. In this study, Acetobacter xylinum (ATCC 700178) was grown in the production medium with different concentrations of carboxylmethylcellulose (CMC) and were evaluated for BC production by using a PCS biofilm reactor. The results demonstrated that BC production was enhanced to its maximum (～13 g/L) when 1.5% of CMC was applied, which was 1.7-fold higher than the result obtained from control culture. The major type of the produced BC was also switched from BC pellicle to small pellets. The ratio of BC pellets in suspension increased from 0 to 93%. Fourier transform infrared (FTIR) spectroscopy demonstrated that CMC was incorporated into BC during fermentation and resulted in the decreased crystallinity and crystal size. The X-ray diffraction (XRD) patterns indicated that CMC-BC exhibited both lower crystallinity (80%) and crystal size (4.2 nm) when compared with control samples (86% and 5.3 nm). The harvested BC was subjected to paper formation and its mechanical strength was determined. Dynamic mechanical analysis (DMA) results demonstrated that BC paper sheets exhibited higher tensile strength and Young's modulus when compared with regular paper.     

Helicobacter pylori in Liquid Culture: Evaluation of Growth Rates and Ultrastructure

This study investigated the growth of Helicobacter (H.) pylori in Brucella broth supplemented with either IsoVitaleX (1% vol/vol), hemin (0.1% wt/vol), agar (0.3% wt/vol), or blood agar blocks (1.5% wt/vol agar). IsoVitaleX was found to significantly shorten the lag phase, while hemin inhibited the growth within the first 24 hours but later acted as a growth stimulant. There was a tendency toward stronger growth when blood agar blocks were added to the medium. Subsequent electron microscopic evaluation revealed that cells of H. pylori were attached to blood agar block surfaces. In contrast, the supplementation of Brucella broth with agar did not significantly increase the cell density. When H. pylori was grown in the presence of IsoVitaleX, strongly stainable electron-dense bodies (140–200 nm) were seen in the cytoplasms. Incubation of cultures on rotary shakers at 120 rpm significantly enhanced growth. The addition of glycerol (15% vol/vol) or fetal bovine serum (15% vol/vol) showed good ultrastructural preservation of bacteria with undamaged cell walls and cytoplasmic membranes, and the cytoplasms were ribosome-dense. Cell counts revealed that cultures stored in glycerol or fetal bovine serum had a significantly lower loss in viability when compared with cultures stored without cryopreservatives. Unprotected cells of H. pylori showed on electron micrographs clumping, cell lysis, and flagellar damage. Finally, the survival rates of H. pylori after multiple thawing from storage at −80°C were best in Brucella broth/glycerol, Brucella broth/fetal bovine serum, and Brucella broth without cryopreservative (in descending order). Received: 10 November 1997 / Accepted: 29 January 1998     

Sodium chloride reduces production of curvacin A, a bacteriocin produced by Lactobacillus curvatus strain LTH 1174, originating from fermented sausage

Lactobacillus curvatus LTH 1174, a strain originating in fermented sausage, produces the antilisterial bacteriocin curvacin A. Its biokinetics of cell growth and bacteriocin production as a function of various concentrations of salt (sodium chloride) were investigated in vitro during laboratory fermentations using modified MRS medium. A model was set up to describe the effects of different NaCl concentrations on microbial behavior. Both cell growth and bacteriocin activity were affected by changes in the salt concentration. Sodium chloride clearly slowed down the growth of L. curvatus LTH 1174, but more importantly, it had a detrimental effect on specific curvacin A production (k(B)) and hence on overall bacteriocin activity. Even a low salt concentration (2%, wt/vol) decreased bacteriocin production, while growth was unaffected at this concentration. The inhibitory effect of NaCl was mainly due to its role as an a(w)-lowering agent. Further, it was clear that salt interfered with bacteriocin induction. Additionally, when 6% (wt/vol) sodium chloride was added, the minimum biomass concentration necessary to start the production of curvacin A (X(B)) was 0.90 g (cell dry mass) per liter. Addition of the cell-free culture supernatant or a protein solution as a source of induction factor resulted in a decrease in X(B), an increase in k(B), and hence an increase in the maximum attainable bacteriocin activity.     

Ethanol Production by Thermophilic Bacteria: Physiological Comparison of Solvent Effects on Parent and Alcohol-Tolerant Strains of Clostridium thermohydrosulfuricum

The effects of temperature, solvents, and cultural conditions on the fermentative physiology of an ethanol-tolerant (56 g/liter at 60°C) and parent strain of Clostridium thermohydrosulfuricum were compared. An ethanol-tolerant mutant was selected by successive transfer of the parent strain into media with progressively higher ethanol concentrations. Physiological differences noted in the mutant included enhanced growth, tolerance to various solvents, and alterations in the substrate range and the fermentation end product ratio. Ethanol tolerance was temperature dependent in the mutant but not in the parent strain. The mutant grew with ethanol concentrations up to 8.0% (wt/vol) at 45°C, but only up to 3.3% (wt/vol) at 68°C. Low ethanol concentration (0.2 to 1.6% [wt/vol]) progressively inhibited the parent strain to where glucose was not fermented at 2.0% (wt/vol) ethanol. Both strains grew and produced alcohols on glucose complex medium at 60°C in the presence of either 5% methanol or acetone, and these solvents when added at low concentration stimulated fermentative metabolism. The mutant produced ethanol at high concentrations and displayed an ethanol/glucose ratio (mole/mole) of 1.0 in media where initial ethanol concentrations were ≤4.0% (wt/vol), whereas when ethanol concentration was changed from 0.1% to 1.6% (wt/vol), the ethanol/glucose ratio for the parent strain changed from 1.6 to 0.6. These data indicate that C. thermohydrosulfuricum strains are tolerant of solvents and that low ethanol tolerance is not a result of disruption of membrane fluidity or glycolytic enzyme activity.     

Osmoadaptation Strategy of the Most Halophilic Fungus,Wallemia ichthyophaga,Growing Optimally at Salinities above 15% NaCl

Wallemia ichthyophaga is a fungus from the ancient basidiomycetous genus Wallemia (Wallemiales, Wallemiomycetes) that grows only at salinities between 10% (wt/vol) NaCl and saturated NaCl solution. This obligate halophily is unique among fungi. The main goal of this study was to determine the optimal salinity range for growth of the halophilic W. ichthyophaga and to unravel its osmoadaptation strategy. Our results showed that growth on solid growth media was extremely slow and resulted in small colonies. On the other hand, in the liquid batch cultures, the specific growth rates of W. ichthyophaga were higher, and the biomass production increased with increasing salinities. The optimum salinity range for growth of W. ichthyophaga was between 15 and 20% (wt/vol) NaCl. At 10% NaCl, the biomass production and the growth rate were by far the lowest among all tested salinities. Furthermore, the cell wall content in the dry biomass was extremely high at salinities above 10%. Our results also showed that glycerol was the major osmotically regulated solute, since its accumulation increased with salinity and was diminished by hypo-osmotic shock. Besides glycerol, smaller amounts of arabitol and trace amounts of mannitol were also detected. In addition, W. ichthyophaga maintained relatively small intracellular amounts of potassium and sodium at constant salinities, but during hyperosmotic shock, the amounts of both cations increased significantly. Given our results and the recent availability of the genome sequence, W. ichthyophaga should become well established as a novel model organism for studies of halophily in eukaryotes.     

Carbohydrate Storage in the Entomopathogenic Fungus Beauveria bassiana

The entomopathogenic fungus Beauveria bassiana was grown in 1% (wt/vol) gelatin-liquid media singly supplemented with a monosaccharide (glucose or fructose), a disaccharide (maltose or trehalose), a polyol (glycerol, mannitol, or sorbitol), or the amino sugar N-acetyl-d-glucosamine. The relative contributions of the carbohydrate, protein, and water contents in the fungal biomass were determined. Carbohydrates composed 18 to 42% of the mycelial dry weight, and this value was lowest in unsupplemented medium and highest in medium supplemented with glucose, glycerol, or trehalose. Biomass production was highest in liquid cultures supplemented with trehalose. When liquid cultures were grown in medium supplemented with 0 to 1% (wt/vol) glucose, trehalose, or N-acetyl-d-glucosamine, there was an increase in the biomass production and the contribution of carbohydrate to mycelial dry weight. Regardless of the glucose concentration in the culture, water content of the mycelia remained about 77.5% (wt/wt). Mycelial storage carbohydrates were determined by capillary gas chromatography. In gelatin-liquid medium supplemented with 1% (wt/vol) glucose, B. bassiana stored glycogen (12.0%, wt/dry wt) and the polyols mannitol (2.2%), erythritol (1.6%), glycerol (0.4%), and arabitol (0.1%). Without glucose, B. bassiana stored glycogen (5.4%), mannitol (0.8%), glycerol (0.6%), and erythritol (0.6%) but not arabitol. To our knowledge, this is the first report of carbohydrate storage in an entomopathogenic fungus, and the results are discussed in relation to other fungi and the potential implications to commercial formulation and insect-fungus interactions.