Persistence of <Emphasis Type="Italic">Isaria fumosorosea</Emphasis> (Hypocreales: Cordycipitaceae) SFP-198 Conidia in Corn Oil-Based Suspension

Long-term persistence of entomopathogenic fungi as biopesticides is a major requirement for successful industrialization. Corn oil carrier was superior in maintaining germination rates of Isaria fumosorosea SFP-198 conidia during exposure to 50°C for 2 h, when compared with other oils, such as soybean oil, cottonseed oil, paraffin oil, and methyl oleate. The corn oil-based conidial suspension (91.6% germination) was also better in this regard than conidial powder (28.4% germination) after 50°C for 8 h. Long-term storage stabilities of corn oil-based conidial suspension and conidial powder at 4 and 25°C for 24 months were investigated, based on the correlation of germination rate with insecticidal activity against greenhouse whiteflies, Trialeurodes vaporariorum. Viability of conidia in corn oil was more than 98.4% for up to 9 months of storage at 25°C, and followed by 23% at 21 months. However, conidial powder had only 34% viability after 3 months of storage at 25°C, after which its viability rapidly decreased. The two conidial preparations stored at 4°C had better viabilities than those at 25°C, showing the same pattern as above. These results indicate that corn oil-based conidial suspension can be used to improve conidial persistence in long-term storage and be further applied to the formulation of other thermo-susceptible biological control agents.     

Conidial water affinity is an important characteristic for thermotolerance in entomopathogenic fungi

The interspecific thermotolerance of several species of entomopathogenic fungi was evaluated based on the conidial water affinity. The species were divided between hydrophilic and hydrophobic conidia. The species with hydrophobic conidia were Beauveria bassiana (ARSEF 252), Metarhizium brunneum (ARSEF 1187), Metarhizium robertsii (ARSEF 2575), Isaria fumosorosea (ARSEF 3889) and Metarhizium anisopliae s.l. (ARSEF 5749). The species with hydrophilic conidia were Tolypocladium cylindrosporum (ARSEF 3392), Tolypocladium inflatum (ARSEF 4877), Simplicillium lanosoniveum (ARSEF 6430), Lecanicillium aphanocladii (ARSEF 6433), S. lanosoniveum (ARSEF 6651), Aschersonia placenta (ARSEF 7637) and Aschersonia aleyrodis (ARSEF 10276). The conidial surface tension of each isolate was also studied. Conidial suspensions were exposed to 38, 41 or 45 °C. After exposure, the suspensions were inoculated on media and conidial germination was evaluated. Considerable differences in thermotolerance were found among the 12 entomopathogenic fungal species. Species with hydrophobic conidia were generally more thermotolerant than species with hydrophilic conidia. All isolates with hydrophobic conidia showed higher conidial surface tension than the isolates with hydrophilic conidia.     

Solid cultures of thrips-pathogenic fungi Isaria javanica strains for enhanced conidial productivity and thermotolerance

Entomopathogenic fungi have great potential to control agricultural and horticultural insect pests, however optimizing conidial production systems to demonstrate high productivity and stability still needs additional efforts for successful field application and industrialization. Although many virulent entomopathogenic fungal isolates have been viewed as potential candidates in a laboratory environment, very few of the isolates are being used in practice for application in agricultural fields as commercial products. I. javanicus is an entomopathogenic fungus that is parasitic to various diverse coleopteran and lepidopteran insects and thought good candidate as biopesticdes. In this work, the basic characteristics of two entomopathogenic fungi, I. javanica FG340 and Pf04, were investigated in morphological examinations, genetic identification, and virulence against Thrips palmi, and then the feasibility of various grains substrates for conidial production was assessed, particularly focusing on conidial productivity and thermotolerance. Isaria javanica FG340 and Pf04 conidia were solid-cultured on 12 grains for 14?days in a Petri dish. Of the tested Italian millet, perilla seed, millet and barley-based cultures showed high conidial production. The four-grain media yielded >1?×?10⁹ conidia/g of I. javanica FG340 and Pf04. Pf04 strain had enhanced thermotolerance up to 45?°C when cultured on Italian millet. In application, it was easy to make a conidial suspension using the cultured grains, and several surfactants were tested to release the conidia. This work suggests several possible inexpensive grain substrates by which to promote conidial production combined with enhanced stability against exposure to high temperature.     

Downstream processing of Beauveria bassiana and Metarhizium anisopliae-based fungal biopesticides against Riptortus pedestris: solid culture and delivery of conidia

We established a fungal production platform by focusing on substrates of solid culture for conidial productivity and thermotolerance, and focusing on surfactants to effectively deliver fungal conidia to the Riptortus pedestris cuticles. First, to produce thermotolerant fungal conidia, 2 of each Beauveria bassiana and Metarhizium anisopliae isolates were cultured on 13 cereal substrates for 10 days. Overall, five substrates (millet, non-glutinous Italian foxtail millet, barley, glutinous Italian foxtail millet, and brown rice) produced greatest number of conidia with thermotolerant conidia. When the selected substrates were mixed with minerals, zeolite, perlite or vermiculite to reduce the amount of cereal grains, vermiculite combination showed relatively high conidial production compared to the other mineral combinations. Next, to efficiently deliver the fungal conidia to the cuticles of R. pedestris, six surfactants, CO-2.5, CO-12, LE-7, PE-61, TED-3, and siloxane were each combined with the fungal conidia. The 0.01% combination showed significantly increased insect mortality, which varied depending on isolate. Virulence tests against R. pedestris were performed with conidial suspensions of isolates to assess their virulence. As a result, isolates showed the highest virulence when a virulence test was conducted at 25°C, rather than 20°C, 30°C and 35°C. This work suggests that the combination of cereal grain substrates and vermiculite could be considered for economic conidial production with high thermotolerance, and the CO-12 surfactant is the most suitable for effective delivery to target insects, followed by the information on optimal temperature for virulence against R. pedestris.     

Effects of culture media on hydrophobicity and thermotolerance of Bb and Ma conidia, with description of a novel surfactant based hydrophobicity assay

Hypocrealean entomopathogenic fungal conidia are made up of multi-aged groups given their chronological conidiogenesis. Most thermotolerance assays have been conducted using mixed-age conidia. The present work exploited a polysiloxane polyether copolymer (siloxane) (Silwet L-77®) mediated conidial collection method, validated by a hydrophobicity assay. This was done to divide mixed-age conidia into two groups based on hydrophobicity and test their thermotolerance, relying on the relationship of conidial age with hydrophobicity. Beauveria bassiana GHA and ERL1170 and Metarhizium anisopliae ERL1171 and ERL1540 conidia, produced on millet agar, whey permeate agar, and ¼SDAY were subjected to hydrophobicity assays that included data on yield of conidia/unit of surface area. Conidia were also collected using 0.01% siloxane, and those remaining with 0.08% siloxane. Hydrophobicity was correlated with percent conidia collected in the two siloxane solutions and yield, suggesting a relationship between percent conidia collected and conidial age (maturation). The conidial suspensions were exposed to 45 °C for 45 min, and conidial germination was examined. Overall, conidia which were collected in 0.08% siloxane had lower germination after heat exposure than those collected in the 0.01% solution. Conidia of both fungi produced by incubation on millet or whey permeate for 14 d were more hydrophobic and exhibited greater thermotolerance than those produced on ¼SDAY. These results suggest that conidia can be divided into two groups with different thermotolerance by using a siloxane-mediated conidial collection method based on hydrophobicity. This depends on the types of substrates used that could influence conidial maturation.     

Generating thermotolerant colonies by pairing Beauveria bassiana isolates

Low thermotolerance in entomopathogenic fungi is the main impediment to their industrialization. This research, for the first time, describes the generation of a thermotolerant colony by pairing and subculturing (cycling) two Beauveria bassiana isolates without sexual reproduction. A mixture of B.?bassiana ERL1578 and ERL1576 was inoculated on quarter-strength Sabouraud dextrose agar with yeast extract (?SDAY). The paired culture (ERL1578?+?1576) was cycled three times to increase the frequency of possible hyphal fusion at the first cycle (c.?5/5?×?10(5) conidia), followed by a heat treatment as a selection pressure. Two non-paired isolates served as controls. Two morphologically different colonies (BbHet1 and BbHet2) were isolated from the pairing. BbHet1 colony had the highest conidial yield. BbHet2 had the most rapid mycelial growth and produced sponge-like mycelial masses (the others were flat), and its conidia were darker than the non-paired colonies under a microscope (400×). BbHet2 conidia had 60.7% germination after exposure to 45?°C for 60?min (the others had 相似文献   

Production of thermotolerant entomopathogenic fungal conidia on millet grain

Thermotolerance of entomopathogenic (insect-killing) fungi should be seriously considered before industrialization. This work describes the feasibility of millet grain as a substrate for production of thermotolerant Beauveria bassiana (Bb) GHA and ERL1170 and Metarhizium anisopliae (Ma) ERL1171 and ERL1540 conidia. First, conidial suspensions of the Bb isolates, produced on millet grain in polyethylene bags, were exposed to five temperatures (43–47°C) at 15-min intervals for up to 120 min (experiment I). Agar-based quarter-strength (¼) Sabouraud dextrose agar supplemented with yeast extract (SDAY) and whey permeate media served as controls. Millet-grain-based culture was superior in producing the most thermotolerant Bb conidia, followed by whey permeate agar and ¼SDAY-based cultures. Secondly, to compare the thermotolerance of conidia produced at the same conditions, the Bb isolates were then produced on agar-based millet powder medium, with ¼SDAY and whey permeate agar media as controls, and the two Ma isolates were added (experiment II). They were then exposed to the same temperatures as above. More thermotolerant Bb and Ma conidia were produced on millet powder agar than on whey permeate agar and ¼SDAY overall. These results suggest that millet grain can be used as a substrate to produce thermotolerant conidia in a mass production system.     

Potential application of the entomopathogenic fungus, shape Nomuraea rileyi, for control of the corn earworm, shape Helicoverpa armigera

The entomopathogenic fungus, Nomuraea rileyi, caused 90.5–100% mortality in fourth-instar larvae of the corn earworm, Helicoverpa armigera, when applied at 10⁷ conidia/ml to corn silks, and leaves of soybean, tomato and chrysanthemum. The LT₅₀ was 5.9–6.7 days. The fifth-instar larvae showed a mortality of 94.6% on soil with 20% water content, and 41.7% on 10% water content when the soil surface was sprayed with 10⁸ conidia/ml suspension. Five fungicides, eight insecticides and nine herbicides, which are commonly used in corn fields, were evaluated for inhibition to conidial germination by a paper disk test. Among them, only two fungicides, viz., maneb and propineb, were highly inhibitory, while insecticides and herbicides examined were not inhibitory to the fungus. Field applications of N. rileyi conidial suspension to neonate larvae were found to be as effective as 40.46% carbofuran (EC) at 800-fold dilution in controlling corn earworm based on marketable ears. It is thus suggested that N. rileyi has potential to be a microbial control agent for this insect.     

Cold activity of Beauveria and Metarhizium, and thermotolerance of Beauveria

Heat and cold are environmental abiotic factors that restrict the use of entomopathogenic fungi as agents for biological control of insects. The thermotolerance and cold activity of 60 entomopathogenic fungal isolates, including five species of Beauveria and one isolate of Engyodontium albus (=Beauveria alba) were examined as to tolerance of temperatures that might be encountered during field use. In addition, cold activity of eight Metarhizium spp. isolates was evaluated. The isolates were from various geographic regions, arthropod hosts or substrates. High variability in conidial thermotolerance was found among the Beauveria spp. isolates after exposure to 45 °C for 2 h, as evidenced by low (0-20%), medium (20-60%), or high germination (60-80%). The thermal death point (0% germination) for three rather thermotolerant B. bassiana isolates (CG 138, GHA and ARSEF 252) was 46 °C for 6 h. At low temperatures (5 °C), with few exceptions (e.g. CG 66, UFPE 479, CG 227, CG 02), most of the B. bassiana isolates germinated well (ca. 100%). On the other hand, only one isolate of Metarhizium sp. was cold-active (i.e. ARSEF 4343 from Macquarie Island, 54.4°S, Australia). This probably is a M. frigidum isolate. The E. albus isolate (UFPE 3138) was the most susceptible isolate to both heat and cold stress. Isolates ARSEF 252 and GHA of B. bassiana, on the other hand, presented exceptionally high thermotolerance and cold activity. Some isolates with high cold activity, however, were thermosensitive (e.g. ARSEF 1682) and others with high thermotolerance had low cold activity (e.g. CG 227). An attempt to correlate the latitude of origin with thermotolerance or cold activity indicated that B. bassiana isolates from higher latitudes were more cold-active than isolates from nearer the equator, but there was not a similar correlation for heat.     

Plant oils for improving thermotolerance of Beauveria bassiana

Conidia of Beauveria bassiana ARSEF-7060 produced in millet amended with plant oils such as sunflower, corn, or cotton seed oil, were exposed to 45 degrees C of wet heat for 90 min. Conidia from millet+corn oil medium had the highest thermotolerance (LT50 (median survival time): 45.7 min). The mycotized millet grains were coated with each of the same plant oils as a granular formulation and subjected to 50 degrees C of dry heat for 8 h. Corn oil coating (LT50: 8.68 h) was superior to sunflower- and cotton seed oil coatings, suggesting the feasibility of using corn oil to increase conidial thermotolerance.     

Ion-beam and gamma-ray irradiations induce thermotolerant mutants in the entomopathogenic fungus Metarhizium anisopliae s.l.

Entomopathogenic fungi, such as Metarhizium anisopliae (Metch.) Sorokin, are important agents for the biological control of insect pests. However, these fungi are not compatible with high temperatures. In this study, mutagenesis using ion beams or gamma rays was used to generate five potentially thermotolerant mutants from two wild-type isolates of M. anisopliae (two using ion beams and three using gamma rays). The mutant isolates had a higher upper thermal limit for vegetative growth compared to the wild types (by 2–3°C) and enhanced tolerance to wet–heat stress of 45°C for conidial germination. At 25°C and 30°C, most mutants were as virulent to maize weevil adults as the wild type, however, one mutant produced using ion beams almost lost virulence entirely. These results indicate that ion beams and gamma rays are useful tools for improving biological characteristics, such as thermotolerance, in entomopathogenic fungi, but that mutants must be carefully evaluated for unpredictable negative side effects.     

Medium components and culture conditions affect the thermotolerance of aerial conidia of fungal biocontrol agent Beauveria bassiana

AIMS: To produce more thermotolerable conidia of Beauveria bassiana, a well-known fungal biocontrol agent, by optimizing the medium components and culture conditions. METHODS AND RESULTS: The conidia produced on media including 0.5-6% glucose, sucrose or starch as carbon source and 50-300-microg ml(-1) Cu2+, Zn2+, Mn2+ or Fe3+ as additive to Sabouraud dextrose medium at 15-30 degrees C, pH 4-8 or KCl-adjusted water availabilities were exposed to 30-min wet heat stress at 48 degrees C. The medium components for conidial production with greatly enhanced thermotolerance included 4% glucose as optimum or 1% starch as alternative for the carbon source and < or =50-microg ml(-1) Mn2+ for the metal additive. The culture conditions were optimized as 25 degrees C and pH 5-6. Conidial thermotolerance decreased remarkably when sucrose and Fe3+ or Cu2+ were used in the cultures, but altered slightly when 50-200-microg ml(-1) Zn2+ were included. CONCLUSIONS: The tolerance of B. bassiana conidia to the thermal stress was significantly affected by the medium composition and culture conditions under which the conidia were produced. SIGNIFICANCE AND IMPACT OF THE STUDY: Proper treatment of small grains as mass production substrates for more glucose release and supplement of glucose or 50-microg ml(-1) Mn2+ are possible means to enhancing conidial thermotolerance and field persistence for improved insect control.     

Increased heat tolerance afforded by oil-based conidial formulations of Metarhizium anisopliae and Metarhizium robertsii

The thermotolerance of oil-based conidial formulations of Metarhizium anisopliae s.l. (IP 46) and Metarhizium robertsii (ARSEF 2575) were investigated. Conidia of IP 46 or ARSEF 2575 were suspended in different adjuvants and exposed to 45?±?0.2°C for 4, 6, 8 or 24?h; their viability was then assessed after 48?h incubation at 27?±?1°C. Conidia heated in pure mineral or vegetable oil exhibited mean relative viability exceeding 70% after 8?h of heat exposure, whereas low germination (≤20%) was observed when conidia were heated in water (Tween 80^® 0.01%), carboxymethyl cellulose gel or emulsifiable oils (Graxol^® or Assist^®) and exposed to heat for 6 or 8?h. In addition, conidia of IP 46 suspended in either pure mineral or canola oil and exposed to heat for 48?h had moderate viability, 57% or 41%, respectively. Unstable oil-in-water emulsions showed a higher percentage of conidia incorporated into oil micellae, while the stable emulsions had higher percentage of conidia outside the oil micellae. The thermotolerance of conidia formulated in stable emulsions, however, did not differ from that of conidia formulated in unstable emulsions. The present study highlights possibilities to alleviate the deleterious effects of heat stress towards Metarhizium spp. conidia applied for controlling arthropod pests and vectors through oil-based formulations.     

The enhanced infectivity of Metarhizium flavoviride in oil formulations to desert locusts at low humidities

Formulation in non-evaporative diluents such as oils is required to exploit the advantages of controlled droplet application methods for biopesticides based on conidial suspensions of entomopathogenic fungi. In laboratory assays on desert locusts (Schistocerca gregaria), formulations of Metarhizium flauoviride conidia in cotton seed oil showed superior performance to water-based suspensions and this was especially pronounced at low humidities (35% r.h.). The LD₅₀s for oil and water suspensions at 5 days were 8900 and > 10⁶ conidia/insect respectively. The dose-mortality regression line for oil was more than four times steeper and median lethal times were typically 76% of those for aqueous suspensions. Formulation in oil improves the efficacy of this fungus and may extend the application of species with lipophilic conidia into less humid agricultural environments.     

Effect of sunscreens,irradiance and resting periods on the germination ofMetarhizium flavoviride conidia

Chemical sunscreens were incorporated into oil formulations of conidia of two isolates of the entomopathogenic fungusMetarhizium spp. and the formulations exposed to simulated solar radiation. After 2 h exposure several sunscreens gave protection as demonstrated by conidial germination after 24 h incubation on gelatin plates at 26°C but only the formulations with Eusolex 8021 showed higher germination than the unprotected control after 48 h incubation. During 5h of exposure, Eusolex 8021 failed to offer significant protection as demonstrated by conidial germination after 48 h incubation. Conidial damage was proportional to the duration of radiation received. Allowing periods of darkness between exposures did not result in decreased loss of viability. Storing conidia, after exposure to simulated radiation, for 24 h prior to germination reduced their viability.     

Relationship between thermotolerance and hydrophobin-like proteins in aerial conidia of Beauveria bassiana and Paecilomyces fumosoroseus as fungal biocontrol agents

AIMS: This study was to illustrate the relationship between the thermotolerance and the contents of hydrophobin-like or formic-acid-extractable (FAE) proteins in aerial conidia of Beauveria bassiana and Paecilomyces fumosoroseus produced on rice-based substrate. METHODS AND RESULTS: Survival indices of 11 isolates were separately assessed as a ratio of the viability of conidia after 3-150 min thermal stress at 48 degrees C over that of unstressed conidia and fitted well to a survival model (r(2) >/= 0.97). For a given isolate, the fitted model generated an LT(50), the time for 50% viability loss under the stress. The LT(50)s of six B. bassiana isolates (10.1-61.9 min) and five P. fumosoroseus isolates (2.8-6.2 min) were correlated (r(2) = 0.81) with FAE protein contents (6.9-23.4 microg mg(-1)). The survival indices of a fixed B. bassiana isolate after 45-min thermal stress at 48 degrees C were also correlated to the FAE protein contents from conidia produced on glucose-, sucrose-, or starch-based substrate (0.79 相似文献   

Performance of two isolates of Isaria fumosorosea from hot climate zones in solid and submerged cultures and thermotolerance of their propagules

Isaria fumosorosea frequently causes mycosis of agricultural pests in the hot semiarid and dry tropical regions of Mexico. Because temperature tolerance restricts the use of fungal biopesticides, we investigated two isolates from these areas for possible development into mycoinsecticides for use in hot weather agricultural zones. We studied the effects of culture system (solid or submerged cultures) and temperature on the fungal growth, extracellular enzyme production, pathogenicity, and thermotolerance of the produced propagules. Between 20 and 28 °C, the specific growth rates of the isolate PCC were higher on solid media, but in the submerged culture, the isolate P43A grew faster even at temperatures of up to 34 °C. On solid media, P43A produced 1.5-fold more proteases than PCC, but in the submerged culture, both strains had similar activities. Under the same culture conditions, PCC produced a blastospore:conidia ratio of 1:2, and P43A produced a ratio of 1:5. PCC aerial conidia had the shortest Lethal Time 50 (LT₅₀, the time to reach 50 % mortality) against Galleria mellonella larvae, but LT₅₀ was equal for the aerial conidia and the submerged propagules of P43A and PCC. The submerged and aerial propagules of P43A were more thermotolerant than those of PCC. Each isolate performed differently in each culture system, and we concluded that the intended production method should be included as a criterion for screening of entomopathogenic fungus. We found that thermotolerance is a specific characteristic of an isolate from a given species. Because of its specific characteristics, P43A shows more promise for the development of a submerged conidia-based mycoinsecticide for foliar application in aqueous form in hot climate regions.     

Impact of moisture and UV degradation on Beauveria bassiana (Balsamo) Vuillemin conidial viability in turfgrass

Beauveria bassiana conidial viability in turfgrass was evaluated using a two-component nucleic acid stain and fluorescence microscopy. Turfgrass samples along with the top 5 cm of soil were used for conidial extraction and viability evaluation on 1, 2, 3, 7, 14 and 21 days after treatment. There were no differences in conidial viability between two Orthoptera strains, 3622 and 5977, and both strains were able to persist in a sandy loam soil for up to three weeks after application. High and low irrigation levels were applied to each of the two strains and results show that higher irrigation (5.1 cm/week) maintains conidial viability better than a low irrigation level (2.5 cm/week). Mean conidial viability was approximately 8–12% greater in plots with the high irrigation regime. Rather than significantly increase soil moisture levels, it is hypothesized that the greater amount of irrigation helps to move the conidia deeper into the thatch layer and soil profile, an area that provides protection from damaging surface temperatures and UV exposure. Rainfall that occurred during the beginning of the 2005 test minimized the irrigation effect, and the irrigation treatment differences were more pronounced after 48 h. Four different UV protectants were evaluated for an impact on conidial viability of strain 3622. Two protectants, an optical brightener and magnesium silicate clay, when added to an emulsifiable oil formulation, significantly increased conidial viability on all evaluation dates. The clay particles act as a sunlight blocker while the optical brightener absorbs UV light. The combined protection from the oil and the additive increased conidial viability by approximately 10% on all evaluation dates. Results from this study provide insight into ways to increase entomopathogenic fungal viability under field conditions.     

基于分生孢子热胁迫反应的球孢白僵菌耐热菌株筛选

将寄主和原产地不同的16株球孢白僵菌Beauveriabassiana的分生孢子分别悬浮于1mL萌发液中,接受48℃恒温水浴45min的热胁迫后,再在25℃下振荡培养24h,以残存活孢率作为各菌株对热胁迫反应的耐热力指标。结果显示,供试菌株的残存活孢率在4.6%~87.1%之间变化,相互间差异极显著。热胁迫后有三株菌的残活孢率均在40%以上,耐热力强;四株菌的残存活孢率低于10%,耐热力弱;其余菌株的残存活孢率为11.0%~23.6%,耐热力居中。从上述菌株中挑选耐热力强、居中和弱的菌株各一株(残存活孢率分别为40.8%、15.8%和4.6%),分别置于25℃、30℃及35℃下平板培养,其菌落生长表现明显与耐热力的强弱相关。虽然三菌株在25℃下的菌落生长无显著差异,但只有耐热力强的菌株能在35℃下正常生长并产孢,是罕见的球孢白僵菌耐热菌株。     

The lipolytic activity of Thermomyces lanuginosus strains isolated from different natural sources

The ability of 144 Thermomyces lanuginosus wild strains isolated from biohumus, mushroom and garden composts, decayed leaves, hazelnuts, and raw coffee beans to hydrolyze synthetic (tributyrin, Tween 20, Tween 40, Tween 60, and Tween 80) and natural fatty substrates (sunflower, soybean, rapeseed and corn oil) was evaluated, and whether the lipolytic activity depended on the isolation source determined. All strains incubated at 55 °C on solid media containing 1% synthetic and 15% natural fatty substrates hydrolyzed both types of substrate. Mean lipolytic activity on natural substrates was significantly higher than on synthetic substrates. The highest mean activity index was noted after growth on sunflower oil, followed by soybean oil and tributyrin; indices on other fatty substrates were low. Strains isolated from raw coffee beans showed the highest mean index, followed by those from biohumus and garden compost; the lowest index being for strains isolated from hazelnuts. Thus, the lipolytic activity index depended on the specific fatty substrate and the source of the isolates.