Enhanced ovicidal activity of an oil formulation of the fungus Metarhizium anisopliae on the mosquito Aedes aegypti

Abstract.  The effect of humidity on the activity of Metarhizium anisopliae IP 46 (Metsch.) Sorokin (Hypocreales: Clavicipitaceae) formulated in sunflower oil against Aedes aegypti (L.) (Diptera: Culicidae) eggs was examined. After exposure of eggs at 75% relative humidity (RH) for ≤ 25 days, ovicidal activity was not increased by oil-in-water formulated conidia, hyphal bodies or pure-oil formulated conidia, compared with conidia or hyphal bodies prepared in water only. At optimal > 98% RH, eclosion was ≤ 13.7% after treatment with oil-in-water formulated propagules in ≤ 10% oil, and it was completely inhibited when conidia were applied in pure oil. At 86–100% RH, new conidia were found on eggs treated with oil-formulated conidia and incubated down to 91% RH. Ovicidal activity was still detected at 93% RH and was augmented with increasing humidity and time of exposure of eggs. Eclosion of larvae was distinctly reduced by IP 46 pure-oil formulated conidia after a minimal initial exposure of 3 days at > 98% RH, followed by: (a) a 12-day exposure at 75% RH before submersion in water; (b) a minimal 5-day exposure at > 98% RH and direct subsequent transfer of treated eggs to water, or (c) a minimal daily 20-h exposure at > 98% RH alternating with 4 h at 75% RH for 10 days. We demonstrate that oil-based formulations of conidia of M. anisopliae enhance ovicidal activity at high humidities and conclude that these formulations have potential in the integrated control of Ae. aegypti .     

Impact of moisture on in vitro germination of Metarhizium anisopliae and Beauveria bassiana and their activity on Triatoma infestans

The in vitro germination of 11 Metarhizium anisopliae and 11 Beauveria bassiana isolates originating from substrates collected in rural peridomestic areas in Central Brazil where triatomines are common was tested. Conidia completed germination up to 24 h after exposure to water activity of >0.99 a_w in all isolates tested. At lower 0.93 a_w germination was delayed but conidia of most isolates germinated at high rates (>98 %) within 216 h of incubation. Activities of 2 M. anisopliae and 2 B. bassiana isolates with different patterns of germination at 0.93 a_w were tested in Triatoma infestans third instar nymphs. There was no relationship between germination kinetics in vitro at 0.93 a_w and their activity in vivo at 98, 75 and 43 % relative humidity (rh). Isolates with accelerated germination at 0.93 a_w were not more virulent at 75 and 43 % rh compared with isolates with retarded or no germination. Highest mortalities were observed at 98 % rh, and they did not exceed 25 % after 25 d incubation at lower 75 and 43 % rh. Isolates that originated from a region with an extensive annual arid period showed no adaptation to lower humidity in their activity against T. infestans.     

Potential of oil-based formulations of Beauveria bassiana to control Triatoma infestans

The in vitro development of Beauveria bassiana conidia was monitored when immersed in six concentrations of seven non-ionic (MP 6400, MP 600, Renex 60, Renex 95, Span 80, Tween 20 and Tween 80) and three anionic (DOS 75, Hostapaval BVQ 9 and Surfax 220) surfactants and 11 vegetable oils (linseed, soybean, groundnut, rapeseed, thistle, sunflower, olive, sesame, corn, castor, and babassu). The influence of the oils on the settling behavior of Triatoma infestans nymphs and the activity of an oil–water formulation of the fungus against this vector under laboratory and simulated field conditions were also determined. With exception of DOS 75 and Surfax 220 germination of conidia on complete medium was >98% at 24 h after exposure to surfactants up to 10%. Elevated rates of germination (>25%) were observed in 10% corn, thistle and linseed oil 8 days after incubation. Pure oils had a significant repellent effect to T. infestans. Repellency decreased generally at 10% of the oil and some oils showed some attractiveness for nymphs when tested at 1%. Nymphs were highly susceptible to oil–water formulated conidia, even at unfavorable moisture for extra-tegumental development of the fungus on the insect cuticle.     

Effectiveness of Metarhizium anisopliae formulations against dengue vectors under laboratory and field conditions

The oviposition behaviour of Aedes aegypti and the effectiveness of Metarhizium anisopliae conidia formulated in water or oil-in-water against A. aegypti adults and eggs were tested in multi-choice and no-choice tests in oviposition devices under laboratory conditions. Both females and males rested in the devices, regardless of the formulation, and were not repelled by the presence of conidia (up to 10⁶?conidia/cm²) without oil or formulated with oil on treated filter paper arranged in the device. However, at higher oil concentrations (≥0.1?μl/cm²), regardless of the presence of conidia, the number of eggs laid by gravid females on the filter paper dropped. The susceptibility of adults, especially of males, to fungal infection increased up to a 15-day incubation. An elevated number of larvae (≥41%) eclosed from eggs laid on the moistened filter paper in the device even without submersion of eggs in water, and these larvae subsequently died. In the laboratory, 1?μl/cm² oil combined with 10⁶?conidia/cm² clearly reduced eclosion to 1.8% after submersion of eggs in water compared to ≥13% eclosion in the control. In field tests in Goiânia, Brazil, eclosion of aedine larvae from eggs laid on filter paper previously treated with oil-in-water formulated conidia dropped to between 0% and 36% compared to 22–50% in the control. Promising results of laboratory and field tests with M. anisopliae formulated in water or oil-in-water and tested in a device emphasised the effectiveness of a fungus-based formulation for aedine mosquitoes in peridomestic areas.     

The mechanism of the mycoinsecticide diluent on the efficacy of the oil formulation of insecticidal fungus

Adverse conditions, including low humidity, UV irradiation, and high temperature, appreciably affect the efficacy of mycoinsecticides. Oil formulation increased the virulence of Metarhizium anisopliae var. acridum (Ascomycota: Hypocreales) against locusts and grasshoppers by reducing the dependence on saturated water. A mycoinsecticide diluent (a water-in-oil emulsion) has been widely used to dilute the oil formulation of M. anisopliae in China. The aim of our study was to elucidate the mechanism by which the mycoinsecticide diluent improves the virulence of M. anisopliae. We investigated the effects of the mycoinsecticide diluent on the virulence, invasion speed, and viability of the conidia under various adverse conditions. The results demonstrated that the mycoinsecticide diluent significantly improved the virulence of conidia at low humidity (68, 75, and 84%). In particular, at an RH of 68%, the LT₅₀ for locusts treated with the emulsion was 5.4 days and was 31.6% lower than the value for locusts treated with an oil formulation. In addition, the concentration of the hyphal bodies found in the haemolymph of locusts treated with emulsion was about 27-fold higher than that in locusts treated with oil formulation four days after inoculation. This result was further confirmed by determining the concentration of M. anisopliae var. acridum DNA in locust haemolymph using quantitative PCR. The percentage germination of conidia in the emulsion was also significantly higher than that in oil at 68% RH. There was no significant difference in percentage germination between conidia treated with the emulsion and oil when exposed to irradiation with ultraviolet-B (UV-B) or high temperature. These results demonstrate that the mycoinsecticide diluent enhances the virulence of M. anisopliae formulated in oil at low humidity by providing adequate water for germination without interfering with the UV tolerance and thermotolerance of the conidia.     

Control of experimental Triatoma infestans populations: effect of pour‐on cypermethrin applied to chickens under natural conditions in the Argentinean Chaco region

Among peridomestic structures, chicken coops are sites of major importance for the domestic ecology of Triatoma infestans (Hemiptera: Reduviidae). The aim of this study was to evaluate in an experimental context the effects of a cypermethrin pour‐on formulation applied to chickens on blood intake, moulting and mortality in T. infestans, under the natural climatic conditions of a region endemic for Chagas' disease. Experimental chicken huts were made of bricks and covered with plastic mosquito nets. Ninety fourth‐instar nymphs were maintained in each hut. The study used a completely random design in which chickens in the experimental group were treated with a cypermethrin pour‐on formulation. Five replicates (= huts) of the experimental and control groups were conducted. The number of live T. infestans, blood intake and moults to fifth‐instar stage were recorded at 1, 5, 20, 35 and 45 days after the application of cypermethrin. Cumulative mortality was higher in nymphs exposed to treated chickens (> 71%) than in control nymphs (< 50%) (P < 0.01). Blood intake and moulting rate were lower in nymphs fed on treated chickens than in control nymphs (P < 0.05). Pour‐on cypermethrin was able to cause significant mortality, although it did not eliminate the experimental population of T. infestans.     

Humidity affects populations of Drosophila suzukii (Diptera: Drosophilidae) in blueberry

Temperature and humidity affect insect physiology, survival, fecundity, reproductive status and behaviour. Complementing previous work investigating the effects of temperature on adult survival and fecundity of the invasive frugivorous pest, Drosophila suzukii (Matsumura), this study was conducted to determine the effect of humidity on D. suzukii larval development, adult survival, fecundity and reproductive status using blueberry as a host substrate. The five constant humidity levels in laboratory bioassays were 20, 33, 71, 82 and 94% RH at 20.6 ± 0.2°C. As RH increased, fecundity and longevity increased. At the higher humidity levels, RH had limited impact on mean generation times (T), larval development and eclosion times. The highest net reproductive rate (R_o = 68) and highest intrinsic rate of population increase (r_m = 0.17) were both recorded at 94% RH. The reproductive status of females, as indicated by the number of mature oocytes per female, was significantly greater at 82 and 94% RH, compared to 71% RH. In addition to the laboratory procedures, we correlated field trap captures over an 81‐day summer period to relative humidity (RH) levels in close proximity to those traps. In the field, low ambient humidity levels resulted in decreased trap captures. A humidity‐dependent population model predicted lower densities of D. suzukii relative to populations at higher humidity. This study supports the hypothesis that cultural practices that minimize lower humidity levels in crops can contribute to the management of D. suzukii. Such methods may include open pruning, drip irrigation and field floor management.     

Management of entomopathogenic fungi in cultures of Tenebrio molitor (Coleoptera: Tenebrionidae)

Larvae of mealworms Tenebrio molitor L. (Coleoptera: Tenebrionidae) have been used as animal feed, but fungal pathogens rapidly downsize the populations, resulting in economic losses. In this work, we established an effective management strategy for fungal pathogens. An entomopathogenic fungus, Beauveria bassiana, was isolated from mealworm cadavers. The bioassay of some isolates of this species at >90% relative humidity revealed that the ERL1575 isolate had the highest virulence. At 20–30% RH, ERL1575 conidia when ingested produced 80% mortality but when sprayed topically produced only <10% mortality. Mealworms that had ingested conidia were exposed to 20, 25, 30 and 35°C and high humidity (>95%) for 5 days. This experiment produced about 90% mortality except at 35°C where mortality was <20%. When 40 fungicides were assayed against ERL1575, fluazinam (1000‐fold) and mancozeb (667‐fold) significantly inhibited conidial germination and/or hyphal growth. When fluazinam and mancozeb were added to the mealworm diet of conidia‐inoculated wheat bran, most were alive 3 days post application. However, 100% mortality resulted 3 days post application in the conidia‐inoculated wheat bran without any fungicides. In conclusion, B. bassiana isolates are pathogenic at <30°C when they are ingested by mealworms but fluazinam and mancozeb can be used for management to control the pathogen in their cultures.     

Laboratory and field studies for the control of Chagas disease vectors using the fungus Metarhizium anisopliae

Chagas disease is one of the most important insect-vectored diseases in Brazil. The entomopathogenic fungus Metarhizium anisopliae was evaluated against nymphs and adults of Panstrongylus megistus, Triatoma infestans, and T. sordida. Pathogenicity tests at saturated humidity demonstrated high susceptibility to fungal infection. The shortest estimates of 50% lethal time (LT₅₀) for P. megistus varied from 4.6 (isolate E9) to 4.8 days (genetically modified strain 157p). For T. infestans, the shortest LT₅₀ was 6.3 (E9) and 7.3 days (157p). For T. sordida, the shortest LT₅₀ was 8.0 days (157p). The lethal concentration sufficient to kill 50% of T. infestans (LC₅₀) was 1.9 × 10⁷ conidia/ml for strain 157p. In three chicken coops that were sprayed with M. anisopliae, nymphs especially were well controlled, with a great population reduction of 38.5% after 17 days. Therefore M. anisopliae performed well, controlling Triatominae in both laboratory and field studies.     

Effects of relative humidity on cocoon formation and survival in the braconid wasp Cotesia glomerata

Abstract The effects of relative humidity (RH) on cocoon formation and survival in the braconid parasitoid wasp Cotesia glomerata (L.) (Hymenoptera: Braconidae) are investigated under various humidity conditions (50, 75, 90, 95 and 100% RH) at 20 °C and under an LD 16 : 8 h photoperiod. The mortality rate at the time of egression from hosts under 100% RH is significantly higher than for other RHs. Cocoon clusters formed at 100% RH spread significantly more than those formed at 50, 75, or 90% RH. Developmental periods differ significantly among RHs under which wasps developed. The mean period from the egression from hosts to adult emergence is 8.7 days when developed at 50–95% RHs, and 8.0 days at 100% RH. The emergence rates of C. glomerata that are maintained under the same humidity conditions after egression from hosts are not significantly different among RHs. However, emergence rates from cocoons that are transferred from 100% RH to 50 and 75% RH are < 70%, although the rates are > 90% in most cases. Some wasps do not emerge from cocoons: more than 60% die after adult eclosion at all RHs; the relative frequency of adult deaths is approximately 90% at 50% RH. Relative humidity influences the cluster and cocoon status strongly: both good clusters and cocoons are formed at low RHs. Emergence rates from cocoons of different ranks are significantly different: the rates of low‐rank cocoons are low at low RHs. The survival of C. glomerata is affected strongly by RH through cocoon formation.     

Effect of relative humidity and origin of isolates of Neozygites tanajoae (Zygomycetes: Entomophthorales) on production of conidia from cassava green mite, Mononychellus tanajoa (Acari: Tetranychidae), cadavers

Neozygites tanajoae is a very specialized fungus pathogenic to the cassava green mite (CGM), Mononychellus tanajoa, an important cassava pest introduced to Africa from the Neotropics. Conidial discharge from cadavers of CGM that died from infections with 14 isolates of N. tanajoae collected from diverse climates of Brazil was quantified to help select potential candidate strains for introduction to Africa. Studies aimed to identify isolates with lower requirements for relative humidity for sporulation and isolates that discharged more conidia during short periods of moisture. At 96 ± 0.5% RH, production of conidia was variable and even isolates from the Brazilian semi-arid region, e.g., Petrolina and Itaberaba, produced few conidia. Significant differences in the numbers of conidia produced by diverse Brazilian isolates were observed after 6, 9 and 12 h at 100% RH. At 100% RH, production of primary conidia increased considerably from an average of 57 ± 4 conidia at 6 h to 509 ± 37 conidia at 12 h. The isolate sporulating least (BIN21) discharged only 45.7% of the number of conidia produced by isolate BIN1, one of the isolates producing the most spores. Results from this study demonstrate that differences in production of conidia among isolates should be considered when selecting Neozygites isolates for new biological control introductions.     

Comparative efficacy of emulsifiable-oil,wettable-powder,and unformulated-powder preparations of Beauveria bassiana against the melon aphid Aphis gossypii

Aphidicidal efficacy of two formulations of Beauveria bassiana strain GHA conidia, an emulsifiable paraffinic oil dispersion (OD) and a clay-based wettable powder (WP), were compared to unformulated conidia (technical material, TC) in laboratory bioassays with adult melon/cotton aphids. For the initial 24 h post-treatment, aphids were incubated under differing humidity conditions: high (100%) versus 'low' (75 ± 3%) relative humidity (RH), and over the subsequent 6 days, all aphids were incubated under the low RH conditions. Mortality from the OD and TC treatments did not differ significantly under any test conditions, and ANOVA revealed no synergism of B. bassiana activity by the OD carrier ingredients. In contrast, formulation as a WP had a small, but significant, negative impact on B. bassiana efficacy. Mean LC50 across formulations was nearly 6.5-fold higher (3457 vs. 539 conidia/mm²) at low vs. high RH. The OD formulation was significantly more effective than the WP, increasing mortality a maximum of 27 percentage points. There were no significant effects of formulation on probit regression slopes. The equivalent slopes and small differences in efficacy of the OD formulation vs. TC preparation support a hypothesis that oils function primarily as spray stickers and spreaders, increasing the efficiency of spray applications.     

Sporulation,storage and infectivity of obligate aphid pathogen Pandora nouryi grown on novel granules of broomcorn millet and polymer gel

Aims: Producing granular cultures of obligate aphid pathogen Pandora nouryi for improved sporulation and storage. Methods and Results: Small millet–gel granules were made of the mixtures of 80–95% millet powder with 5–20% polymer gel (polyacrylamide, polyacrylate or acrylate‐acrylamide copolymer) and inoculated with mycelia at 30 mg biomass g^?1 dry granules plus 87·5% water, followed by static incubation at 20°C for 4–12 days. The fungus grew well on 12 preparations but best on that including 10% copolymer. An 8‐day culture of this preparation discharged maximally 58·5 × 10⁴ conidia mg^?1 granule at 100% RH and was capable of ejecting conidia at the nonsaturated regimes of 86–97% RH. During storage at 6°C, granular cultures with >85% water content had twofold longevity (120 days) and half‐decline period (34–36 days) of those stored at room temperature. The steadily high water content preserved the cultures better than that decreasing at 6°C. However, conidia from 70‐day‐stored granules were less infective to Myzus persicae nymphs than those from fresh ones based on their LC₅₀s. Conclusions: The millet–gel granules had higher sporulation capacity than reported Pandora cultures and a capability of spore discharge at nonsaturated humidity. Significance and Impact of the Study: The granular cultures are more useful for aphid control.     

Effect of relative humidity,temperature and fungicide on germination of conidia of Cercospora canescens caused the Cercospora leaf spot disease in mungbean

The aim of the present investigation was to determine the impact of relative humidity (RH) and temperature on conidial germination, nuclear position and effect of important fungicides on growth and conidial germination of Cercospora canescens. Germination of conidia was observed at RH range 92–100% at 5–35°C. Significant interaction between temperature and RH indicated that higher humidity and high temperature promoted quick germination both in the presence and absence of free moisture. Although in absence of free moisture at 92–95% RH higher temperatures 25–35°C promoted quick evaporation of moisture and no conidial germination. Number of germtube was increased significantly at the optimum temperature 25–30°C and higher humidity (98–100%). But higher temperature 25–35°C with lower RH did not support the conidial germination. This finding is very important for disease forecasting using meteorological data. The spray of Carbendazim as contact fungicide may not be useful since it is not effective against the conidia of C. canescens. Triadimefon did not inhibit the conidia germination but completely inhibited mycelium development at 50 μg/ml. Propriconazole inhibited both conidia germination and mycelial development. Therefore, Propiconazole may be taken as protective as well as curative spray. In non-systemic fungicide, Copper oxychloride gave anticipated result by inhibiting both conidial germination and mycelium development. Therefore, copper oxychloride can be used as protectant fungicides for Cercospora leaf spot caused by C. canescens.     

Studies on Prolonged Storage of Beauveria bassiana Conidia: Effects of Temperature and Relative Humidity on Conidial Viability and Virulence against Chickpea Borer,Helicoverpa armigera

Unformulated conidia of Beauveria bassiana were stored at five different temperatures (0°, 10°, 20°, 30° and 40°C) at six different relative humidities (RH) (0, 33, 53, 75, 85 and 98%). Conidial viabilities and virulence against third instar larvae of Helicoverpa armigera were determined over a 24‐month period. Conidia survived longest at lower temperatures (0–20°C) and lower RH levels (0–53% RH). At higher temperatures (30–40°C) conidia did not survive. When the temperature was decreased from 30°C to 0°C, at nearly all RH levels the longevity of conidia increased. Conidia remained virulent for third instar larvae of H. armigera under favourable storage conditions for 24 months.     

Influence of humidity on the infection of western flower thrips, <Emphasis Type="Italic">Frankliniella occidentalis</Emphasis> (Thysanoptera: Thripidae), by <Emphasis Type="Italic">Beauveria bassiana</Emphasis>

The insecticidal activity of Beauveria bassiana GHA derived from a commercial mycoinsecticide BotaniGard ES against Frankliniella occidentalis was determined in a bioassay by dipping the female adults into a conidial suspension. The 90% lethal concentration of B. bassiana GHA was estimated to be 9.7 × 10⁶ conidia/ml. The lethal times for achieving 90% mortality of thrips inoculated with a 1/500-diluted solution of BotaniGard ES and a 10^7.5 (3.16 × 10⁷) conidia/ml suspension of B. bassiana GHA were estimated to be five and six days, respectively. When the treated thrips were exposed to a high relative humidity (RH) of over 99% for various periods and then transferred to 60% RH, the requisite lengths of the high-humidity period to achieve 90% mortality of the thrips at six days after inoculation were estimated to be 46 and 47 h in BotaniGard ES and B. bassiana GHA, respectively. Fungal multiplication in the thrips was detected between 48 to 60 h after inoculation by measuring Beauveria-specific DNA in the host following inoculation with a B. bassiana GHA suspension of 10^7.5 conidia/ml using a real-time quantitative PCR. The mycelial growth in the host hemocoel was not influenced by the low-humidity condition.     

Different host plant species modifies the susceptibility of Bactericera cockerelli to the entomopathogenic fungus Beauveria bassiana

Bactericera cockerelli (Sulc.) is a serious pest of solanaceous crops and a vector of the plant pathogen Candidatus Liberibacter psyllaurous. Entomopathogenic fungi are the most important biological control alternatives for this pest. Host plant species, however, can modify the outcomes of insect–pathogen interactions. We conducted laboratory experiments to quantify the virulence of two isolates of the entomopathogenic fungus Beauveria bassiana (Bals. [Vuill.]), BB40 and BB42, against third instar B. cockerelli nymphs maintained on chilli pepper plants. Owing to the lack of difference in virulence against B. cockerelli nymphs on chilli pepper between the two B. bassiana isolates, only BB42 was used to: compare virulence against nymphs maintained on either chilli pepper, potato or tomato; and in vivo conidia production from nymphs maintained on different host plants. Virulence of the two B. bassiana isolates against B. cockerelli nymphs was similar. Bactericera cockerelli nymphs maintained on tomato were more susceptible to B. bassiana than nymphs maintained on potato or chilli peppers. Infected nymphs maintained on chilli peppers produced the greatest number of conidia followed by infected nymphs maintained on tomato and potato. Host plant affected the susceptibility of B. cockerelli to B. bassiana isolate BB42 and subsequent conidia production. The implications of our results for microbial control of B. cockerelli by B. bassiana are discussed.     

Impact of moisture on survival of Aedes aegypti eggs and ovicidal activity of Metarhizium anisopliae under laboratory conditions

The effect of relative humidity (43%, 75%, 86% and > 98%) on Aedes aegypti eggs treated with Metarhizium anisopliae or water only was tested for up to a six months exposure at 25 degrees C. Survival of larvae inside eggs was clearly affected by the lowest humidity (43%) tested, and eclosion diminished at all humidities after increasing periods of exposure. M. anisopliae showed to have a strong ovicidal activity only at humidity close to saturation. No difference of activity was found between conidia and hyphal bodies tested. This fungus affected larvae inside eggs and has potential as a control agent of this important vector in breeding sites with high moisture.     

Humidity‐controlled cold storage of Neoseiulus californicus (Acari: Phytoseiidae): effects on male survival and reproductive ability

Humidification can suppress water loss from an organism and has great potential for improving the cold storage of short‐lived arthropods, such as predatory mites. The effectiveness of humidity‐controlled cold storage was recently verified for Neoseiulus californicus (McGregor) females but was not examined for males. Combining both males and females in one storage protocol might increase the predator population because it would enhance the opportunity for multiple mating, which is necessary for females to maximize their egg production. Newly emerged adult males were stored at an air temperature of 5°C and relative humidity (RH) of 100% or 80%. The median survival time (LT₅₀) was 32 days at 100% RH and 14 days at 80% RH; the survival curves differed significantly. Males stored at 100% RH for 0, 10, 20 and 30 days were introduced to virgin females for mating at 25°C to evaluate their reproductive ability. The pre‐oviposition period was significantly prolonged in the females mated with males stored for ≥20 days. No negative effects of storage were observed on the oviposition period, total number of eggs or net reproduction rate (R₀) in the females mated with males stored for ≤20 days or on the mean generation time (T) for those stored for 30 days. A slight decrease in the intrinsic rate of increase (r_m) was observed in the females mated with males stored for ≥20 days. Our storage method can preserve N. californicus males for 20 days with only a minor reduction in their survival and reproductive ability.     

Evaluation of the Entomopathogenic Fungi Beauveria bassiana and Paecilomyces fumosoroseus for Microbial Control of the Silverleaf Whitefly, Bemisia argentifolii

Collaborative research was conducted at the USDA-ARS Subtropical Agricultural Research Center in southern Texas to assess the microbial control potential of Beauveria bassiana and Paecilomyces fumosoroseus against Bemisia whiteflies. Laboratory assays demonstrated the capacity of both pathogens to infect Bemisia argentifolii nymphs on excised hibiscus leaves incubated at relative humidities as low as 25% at 23 ± 2°C (ca. 35% infection by B. bassiana and P. fumosoroseus resulted from applications of 0.6–1.4 × 10³ conidia/mm² of leaf surface). In small-scale field trials using portable air-assist sprayers, applications at a high rate of 5 × 10¹³ conidia in 180 liters water/ha produced conidial densities of ca. 1–2.5 × 10³ conidia/mm² on the lower surfaces of cucurbit leaves. Multiple applications of one isolate of P. fumosoroseus and four isolates of B. bassiana made at this rate at 4- to 5-day intervals provided >90% control of large (third- and fourth-instar) nymphs on cucumbers and cantaloupe melons. The same rate applied at 7-day intervals also provided >90% control in zucchini squash, and a one-fourth rate (1.25 × 10¹³ conidia/ha) applied at 4- to 5-day intervals reduced numbers of large nymphs by >85% in cantaloupe melons. In contrast to the high efficacy of the fungal applications against nymphs, effects against adult whiteflies were minimal. The results indicated that both B. bassiana and P. fumosoroseus have strong potential for microbial control of nymphal whiteflies infesting cucurbit crops.