Host age and pathogen exposure level as factors in the susceptibility of the house fly,Musca domestica (Diptera: Muscidae) to Beauveria bassiana

Adult house flies, Musca domestica L., of four ages, <1, 3, 7, and 14 day post-eclosion, were exposed to three strains of Beauveria bassiana (P89, L90 and 447). Flies were exposed to moistened filter paper treated with either a low (1.57×10⁴ conidia/cm²) or high (1.57×10⁵ conidia/cm²) concentration of each fungal strain for 6 h. Strain 447 was superior to the two house fly-derived B. bassiana strains in inducing host infection and mortality. Significant spikes in infection and mortality occurred as early as 5 days post-exposure with higher concentration exposures acting more quickly. Few differences were observed in either infection or mortality among the four fly age classes. On Day 10 post-exposure, 77% of the high-concentration, 447-exposed flies were infected, compared with only 24% of the flies from the P89 low-concentration exposure. Potential applications of these results in integrated house fly management programs are discussed.     

Infection of Blissus antillus (Hemiptera: Lygaeidae) Eggs by the Entomopathogenic Fungi Metarhizium anisopliae and Beauveria bassiana

This study determined the pathogenicity and virulence of Beauveria bassiana and Metarhizium anisopliae to eggs of the chinch bug Blissus antillus (Hemiptera: Lygaeidae). Eggs were inoculated under laboratory conditions by immersion in concentrations of 1 × 10⁴ and 5 × 10⁶ conidia/ml. Inoculated eggs were kept under controlled conditions. Evaluations were carried out daily for 20 days. M. anisopliae isolates were highly virulent to eggs, even at 1 × 10⁴ conidia/ml. All B. bassiana isolates tested were considered to be of low virulence or avirulent. The most virulent isolate tested was ESALQ 818 (M. anisopliae), which caused 96.7% infection, when eggs were immersed in suspensions of 1 × 10⁴ conidia/ml. Conidial production on infected eggs was observed to be highest for M. anisopliae isolate CG144, with a mean value of 11.6 × 10⁵ conidia/ml/egg. Infection of Blissus eggs oviposited on plant stems was greater when M. anisopliae isolate CG144 was formulated in mineral oil (63.5% mortality) than when formulated in Tween 80 (27.1% mortality).     

Susceptibility of different life stages of the European cherry fruit fly,Rhagoletis cerasi,to entomopathogenic fungi

The effects of six fungus isolates on the mortality of different life stages of the European cherry fruit fly, Rhagoletis cerasi (Diptera: Tephritidae), were assessed in a series of laboratory experiments to find an isolate suitable for biological control. In a first step, the effects of fungus treatments on mortality, mycosis and fecundity of adult flies at a concentration of 10⁷ conidia/ml were evaluated. All fungus isolates caused mycosis but virulence varied considerably among the isolates. Beauveria bassiana and Isaria fumosorosea caused 90–100% mortality and had the strongest influence on fecundity. Metarhizium anisopliae also induced high rates of mortality, while the pathogenicity of Isaria farinosa was low. The effects of lower conidia concentrations and the influence of the age of flies were assessed in a second step. Higher conidia concentrations generally resulted in a higher mortality. B. bassiana was most efficient at low concentrations. Young flies showed lower mortality rates than older flies but, sub‐lethal effects on eclosion rate of eggs were greater in younger flies. Finally, the effects on L₃ larvae were tested: none of the fungus isolates induced mortality in more than 25% of larvae. As L₃ larvae and pupae are not susceptible to fungus infection, field control of R. cerasi should be focused on adult flies.     

Pathogenicity, formulation and storage of insect pathogenic hyphomycetous fungi tested against Diabrotica speciosa

Studies were conducted tosearch for fungal strains with potentialpathogenicity against Diabrotica speciosa(Germar) (Coleoptera: Chrysomelidae).Among sixteen fungal isolates screenedthe most virulent was a Beauveria bassiana(Balsamo) Vuillemin isolate (FHD13) thatcaused 70% mortality of D. speciosathird instar larvae. The LC₅₀ value ofB. bassiana isolate FHD13 was3.48 × 10¹⁰ conidia/ml.Different temperatures (4, 17 and 26 °C)and vegetable oils (corn, sunflower and canola)used for storage did not significantly affectviability of conidia. A pathogenicity trialagainst D. speciosa larvae performed withthe corn oil formulation (1 × 10⁸ conidia/mlof oil) caused 65% of mortality.     

A new microbial consortia containing entomopathogenic fungus,Beauveria bassiana and plant growth promoting rhizobacteria,Pseudomonas fluorescens for simultaneous management of leafminers and collar rot disease in groundnut

The virulence of 20 isolates of Beauveria bassiana (Balsamo) Vuillemin to larvae of the leafminer, Aproaerema modicella, was tested in the laboratory. Leafminer larvae were sprayed with a standard concentration of 1×10⁸ condia/mL of each B. bassiana isolate. All the B. bassiana isolates tested were pathogenic to A. modicella and the mortality varied between 16.7 and 68.9%. Beauveria bassiana isolate B2 was found to be the most virulent followed by isolate B4 which resulted in 59% mortality. Beauveria isolate B2 was selected for dose–response mortality studies with four different doses (1×10², 1×10⁴, 1×10⁶ and 1×10⁸ conidia/mL). Among the various doses tested, 1×10⁸ conidia/mL produced the highest mortality (70.7%). In addition, morphogenesis of the insect pest in all stages, larval, pupal and adult was greatly affected due to fungal infection. Further, B. bassiana isolate B2 and two Pseudomonas fluorescens strains, TDK1 and Pf1 were tested alone and in combination for suppression of groundnut leafminer and collar rot disease and promotion of plant growth and yield both under glasshouse and field conditions. The mixture of B. bassiana and P. fluorescens strains significantly reduced the leafminer and collar rot disease incidences when applied as talc-based formulation through seed, soil and foliar application under glasshouse and field conditions.     

Selection of Beauveria bassiana isolates for control of the whiteflies Bemisia tabaci and Trialeurodes vaporariorum on the basis of their virulence, thermal requirements, and toxicogenic activity

As part of a 3-fold approach to select potential mycoinsecticides for whitefly control, we evaluated infectivity, thermal requirements, and toxicogenic activity of the entomopathogenic fungus Beauveria bassiana (Ascomycota: Clavicipitaceae) under laboratory conditions. Twenty-five native B. bassiana isolates and a commercially available mycoinsecticide (based on B. bassiana) were evaluated for virulence to fourth instar nymphs of sweetpotato whitefly, Bemisia tabaci, and greenhouse whitefly, Trialeurodes vaporariorum, at a concentration of 1 × 10⁷ conidia/ml. All isolates were pathogenic for both whitefly species, whereas mortality rates varied from 3 to 85%. A second series of bioassays was conducted on 10 selected isolates using four 10-fold concentrations ranging from 1 × 10⁵ to 1 × 10⁸ conidia/ml. Median lethal concentrations (LC₅₀) of the four most virulent isolates varied from 1.1 × 10⁵ to 6.2 × 10⁶ conidia/ml and average survival time (AST) of treated nymphs from 5.9 to 7.4 days. T. vaporariorum were significantly more susceptible to all B. bassiana isolates than B. tabaci. The thermal biology of the eight most virulent isolates to both whitefly species was investigated at six temperatures (10–35 °C). The colony radial growth rate was estimated from the slope of the linear regression of colony radius on time and data were then fitted to a modified generalized β function that accounted for 90.5–99.3% of the data variance. Optimum temperatures for extension rate ranged from 23.1 to 27.1 °C, whereas maximum temperatures for fungal growth varied from 31.8 to 36.6 °C. On the basis of their virulence and thermal requirements, three isolates showed promise as candidates for whitefly management in Mediterranean greenhouses. Whilst in vitro production of macromolecular compounds toxic to Galleria mellonella larvae was not a requisite for virulence, ASTs of larvae injected with Sephadex G-25 fractions from candidate isolates ranged from 1.4 to 3.7 days compared with 5–6 days for non-toxic G-25 fractions. In addition, proteinase K treatment significantly reduced their toxic activity suggesting that they were proteins and revealing the potential of these isolates to be further improved through biotechnology to kill the pest more quickly.     

Pathogenicity of Beauveria bassiana and Metarhizium anisopliae to the tobacco spider mite Tetranychus evansi

Seventeen isolates of Metarhizium anisopliae (Metschnikoff) Sorokin and two isolates of Beauveria bassiana (Balsamo) Vuillemin were evaluated for their pathogenicity against the tobacco spider mite, Tetranychus evansi Baker & Pritchard. In the laboratory all the fungal isolates were pathogenic to the adult female mites, causing mortality between 22.1 and 82.6%. Isolates causing more than 70% mortality were subjected to dose–response mortality bioassays. The lethal concentration causing 50% mortality (LC₅₀) values ranged between 0.7×10⁷ and 2.5×10⁷ conidia ml⁻¹. The lethal time to 50% mortality (LT₅₀) values of the most active isolates of B. bassiana and M. anisopliae strains varied between 4.6 and 5.8 days. Potted tomato plants were artificially infested with T. evansi and treated with B. bassiana isolate GPK and M. anisopliae isolate ICIPE78. Both fungal isolates reduced the population density of mites as compared to untreated controls. However, conidia formulated in oil outperformed the ones formulated in water. This study demonstrates the prospects of pathogenic fungi for the management of T. evansi.     

Differential susceptibility of Amblyomma maculatum and Amblyomma americanum (Acari:Ixodidea) to the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae

Nymph and adult ticks from Ambylomma americanum and Ambylomma maculatum were treated with conidia and blastospores of the entomopathogenic fungi Beauveria bassiana (90517) and Metarhizium anisopliae (20500). Fungal suspensions of conidia harvested from potato dextrose plates containing 10⁸ conidia/ml caused greater than 90% mortality in adult A. maculatum but less than 10% mortality in adult A. americanum over a 28 day time course. Similarly, infection with M. anisopliae (10⁸ conidia/ml) resulted in 60 and 15% mortality in A. maculatum and A. americanum, respectively. Nymphs of both tick species were more susceptible to fungal infection reaching mortality rates of almost 100% for A. maculatum and over 35% for A. americanum. Scanning electron microscopy of infected ticks showed rapid attachment, germination, and proliferation of fungal spores on A. maculatum cuticles, and to a much lesser extent on A. americanum cuticles. Pentane extracts of A. americanum cuticle hydrocarbons inhibited germination and hyphal growth of B. bassiana conidia, whereas no inhibition was observed using A. maculatum extracts. Significant mortality towards A. americanum was observed (>60%, 28 days) only when the ticks were treated with B. bassiana directly from the growth medium (10⁷ blastospores/ml, grown for 3–4 days in Sabouraud dextrose + 0.5% yeast extract liquid media). These results indicate tick species display differential susceptibility to the entomopathogenic fungi B. bassiana and M. anisopliae, and that the ability to overcome fungistatic compounds present in the tick epicuticle may determine the likelihood of successful infection and virulence.     

Exchange of Inoculum of Beauveria bassiana (Bals.) Vuill. (Hyphomycetes) Between Adult Flies of the Cabbage Maggot Delia radicum L. (Diptera: Anthomyiidae)

Adult cabbage maggots ( Delia radicum L.) were exposed to dry conidia of isolates of several hyphomycetous fungi by placing them in a centrifuge tube containing conidia, then releasing them into small screened plastic cages. Mortality was assessed after 48, 120 and 160 h. A Beauveria bassiana isolate (P89 from Musca domestica ) caused the highest mortality after 48 h, resulting in 100% mortality and 100% infection. Isolate L90 ( B. bassiana ) and one Metarhizium anisopliae isolate (ARSEF 2521) also caused fatal infection in more than 50% of the flies. To investigate exchange of inoculum, flies were placed in a small container with a dry powder formulation containing B. bassiana (Mycotrol ® ) on the bottom. The flies were removed to small screened cages containing untreated flies. This experiment confirmed the ability of flies to pass inoculum to other flies. In a similar experiment, one treated fly was placed in each cage with one untreated fly. When each fly died, one untreated fly was added to each cage after the dead fly was removed. This study showed that fly to fly transfer of fatal doses of inoculum was possible for a series of at least six flies. When female flies were exposed to the inoculum, then transferred to small cages containing males and an oviposition substrate, no eggs were laid. Further studies are being conducted to develop a system where flies attracted to a trap will be inoculated with the fungus and spread it to a field population.     

Susceptibility of Brevipalpus phoenicis to entomopathogenic fungi

The pathogenicity of 52 isolates from several fungus species was studied for the false spider mite Brevipalpus phoenicis. In addition, the main stages during the course of infection by Hirsutella thompsonii, by far the most virulent pathogen, were studied by means of light and electron microscopy. Adult mites were confined to arenas prepared with citrus leaves in acrylic dishes containing agar–water. Conidial suspensions containing 10⁸ conidia/ml were applied, except for H. thompsonii, where a concentration of 10⁷ conidia/ml was used. The H. thompsonii isolates caused higher mortality, with indices higher than 90%. Observations under the scanning electron microscope (SEM) were performed at 0, 6, 12, 24, 48, 72, and 120 h after application of a H. thompsonii suspension containing 10⁷ conidia/ml. Twenty-four hours after inoculation, H. thompsonii conidia were observed attached to the mite’s integument. The conidia germinated and penetrated through the base of the setae on the hysterosoma. Colonization occurred after 48 h, as evidenced by mortality. Conidiogenesis occurred after 120 h, with the development of mycelium and conidiophores emerging from the posterior and anterior parts of the mite.     

Stage-specific mortality of Rhagoletis indifferens (Diptera: Tephritidae) exposed to three species of Steinernema nematodes

Mortality of larval, pupal, and adult western cherry fruit fly, Rhagoletis indifferens (Tephritidae) exposed to the steinernematid nematodes Steinernema carpocapsae, Steinernema feltiae, and Steinernema intermedium, was determined in the laboratory and field. Larvae were the most susceptible stage, with mortality in the three nematode treatments ranging from 62 to 100%. S. carpocapsae and S. feltiae were equally effective against larvae at both 50 and 100 infective juveniles (IJs)/cm². S. intermedium was slightly less effective against larvae than the other two species. Mortalities of R. indifferens larvae at 0, 2, 4, and 6 days following their introduction into soil previously treated with S. carpocapsae and S. feltiae at 50 IJs/cm² were 78.6, 92.5, 95.0, and 77.5% and 87.5, 52.5, 92.5, and 70.0%, respectively, and at 100 IJs/cm² were 90.0, 92.0, 100.0, and 84.0% and 90.0, 50.0, 42.0, and 40.0%, respectively. There was no decline in mortality caused by S. carpocapsae as time progressed, whereas there was in one test with S. feltiae. Larval mortalities caused by the two species were the same in a 1:1:1 vermiculite:peat moss:sand soil mix and a more compact silt loam soil. In the field, S. carpocapsae and S. feltiae were equally effective against larvae. Pupae were not infected, but adult flies were infected by all three nematode species in the laboratory. S. carpocapsae was the most effective species at a concentration of 100 IJs/cm² and infected 11–53% of adults that emerged. The high pathogenicity of S. carpocapsae and S. feltiae against R. indifferens larvae and their persistence in soil as well as efficacy in different soil types indicate both nematodes hold promise as effective biological control agents of flies in isolated and abandoned lots or in yards of homeowners.     

Susceptibility of Ceratitis capitata Wiedemann (Diptera: Tephritidae) to Entomopathogenic Fungi and Their Extracts

The effectiveness of seven strains of entomopathogenic fungi against Ceratitis capitata adults was evaluated in the laboratory. Adults were susceptible to five of seven aqueous suspensions of conidia. Metarhizium anisopliae and strain CG-260 of Paecilomyces fumosoroseus were the most pathogenic fungi, with 10-day LD₅₀ values of 5.1 and 6.1 × 10³ conidia/fly, respectively, when applied topically. Sublethal effects on fecundity and fertility of the fungal-exposed females were also studied. The most effective fungus in reducing fecundity was P. fumosoroseus CECT 2705, with reductions on the order of 65% at 1 × 10⁶ conidia/fly. M. anisopliae and Aspergillus ochraceus also showed significant reductions of fecundity (40–50% for most of the assayed concentrations). Fertility was moderately affected by the fungi. M. anisopliae at 1 × 10⁶ conidia/fly was the most effective fungus, showing egg eclosion reduction of over 50% compared with the control. In addition, culture broth dichloromethane extracts from the entomopathogenic fungi were tested for insecticide activity against C. capitata, including effects on fecundity and fertility. The extract from M. anisopliae was the most toxic, resulting in about 90% mortality at a concentration of 25 mg/g of diet; under these conditions, fecundity and fertility of treated females were reduced by 94 and 53%, respectively, compared with untreated controls.     

Pathogenicity of the entomogenous,hyphomycete fungus,Metarhizium anisopliae against the chrysomelid beetles Psylliodes chrysocephala and Phaedon cochleariae

Susceptibility of the mustard beetle (Phaedon cochleariae) and the cabbage stem flea beetle (Psylliodes chrysocephala) to six isolates of the entomogenous, hyphomycete fungus Metarhizium anisopliae, was investigated. A farther six isolates were assayed against P. cochleariae only. The isolates originated from hosts of various insect orders. Five of the six isolates tested against P. chrysocephala and P. cochleariae were infective for both species whereas one isolate, V107, was non‐pathogenic to both. The level of virulence of different M. anisopliae isolates for these chrysomelid beetles varied considerably. Isolates V90 and V93 were highly virulent to P. chrysocephala and P. cochleariae respectively but were significantly less virulent against the alternate host species. The LT₅₀ of isolate V90 for P. chrysocephala was 7 days at 4 x 10⁷ conidia/ml and its LC₅₀ value was 16 x 10⁵ conidia/ml. The LT₅₀ of V93 for P. cochleariae was approximately 8 days at 4 X 10⁸ conidia/ml and its LC₅₀ value was 3 x 10⁷ conidia/ml. Following inoculation, germinating conidia of all isolates produced appressoria on the cuticular surface of both hosts suggesting that specificity is determined at later stages of infection.     

Efficacy of Beauveria bassiana against the strawberry pests, Lygus lineolaris, Anthonomus signatus and Otiorhynchus ovatus

There are several insect species causing serious economic losses in strawberry, Fragaria vesca L., productions. In Quebec, Canada, the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), the strawberry bud weevil clipper, Anthonomus signatus (Say) and the strawberry root weevil, Otiorhynchus ovatus (L.) are the most important pests. We tested the susceptibility of these pests to the entomopathogenic fungus Beauveria bassiana under laboratory conditions. Sixteen isolates were evaluated for their insecticide potential against these insects. Adults of each species were infected by the immersion method. All isolates were pathogenic to adults of all three species, causing mortality rates between 23.3% and 100% at a concentration of 1 × 10⁷ conidia/ml. Based on the screening results, isolate INRS‐CFL was selected for its insecticide potential and then used for further analyses against L. lineolaris, A. signatus and O. ovatus adults. Bioassays were performed to evaluate the lethal concentration (LC₅₀) and the average survival time (AST) of this isolate against both insect species. Results of dose–response mortality bioassays using four concentrations – 1 × 10⁴, 1 × 10⁶, 1 × 10⁸ and 1 × 10⁹ conidia/ml – indicated a LC₅₀ values of 5.3 × 10⁵, 1.8 × 10⁷ and 9.9 × 10⁷ conidia/ml at 7 days after inoculation for L. lineolaris, A. signatus and O. ovatus respectively. Using a dose of 1 × 10⁸ conidia/ml, the AST values were estimated at 4.41, 7.56 and 8.29 days, respectively, at a concentration of 1 × 10⁸ conidia/ml. This study demonstrated the potential of B. bassiana for the management of L. lineolaris, A. signatus and O. ovatus. Results also suggest that the heteropteran species is more susceptible than coleopteran species to B. bassiana.     

Comparative laboratory studies on three fungal pathogens of the elm bark beetle Scolytus scolytus: Pathogenicity of Beauveria bassiana,metarhizium anisopliae, and Paecilomyces farinosus to larvae and adults of S. scolytus

The entomogenous fungi Beauveria bassiana (nine isolates), Metarhizium anisopliae (seven isolates), and Paecilomyces farinosus (four isolates) were tested as pathogens of larvae of the elm bark beetle, Scolytus scolytus. Single isolates of B. bassiana and M. anisopliae were also tested against adult beetles. Of the 21 isolates tested as conidial suspensions against larvae, all proved pathogenic. The three most and least virulent isolates were, respectively, isolates of B. bassiana and M. anisopliae. The other isolates fell between these two extremes, with the four P. farinosus isolates all moderately virulent. Spore retention on larvae following inoculation was estimated by washing conidia off the larvae. From the results it was possible to relate larval mortality to the approximate spore dose causing infection at different spore concentrations. Thus, application of spores of the three pathogens at a concentration of 10³ spores/ml resulted in limited mortality. At this concentration, an average of only a single spore was recovered from the inoculated larva. Adult bark beetles also proved susceptible to infection by isolates of B. bassiana and M. anisopliae. They were exposed to discs of elm bark dipped in a conidial suspension. It was estimated that a dose of less than 100 spores could cause infection of beetles following feeding on the elm bark discs.     

Susceptibility of the tobacco whitefly,Bemisia tabaci (Hemiptera: Aleyrodidae) biotype Q to entomopathogenic fungi

Recently, the Q biotype of tobacco whitefly has been recognized as the most hazardous strain of Bemisia tabaci worldwide because of increased resistance to some insecticide groups requiring alternative strategies for its control. We studied the susceptibility of this biotype of B. tabaci to 21 isolates of Beauveria bassiana, three isolates of Isaria fumosorosea, one isolate of I. cateni, three isolates of Lecanicillium lecanii, one isolate of L. attenuatum, and one isolate of Aschersonia aleyrodis. These isolates were evaluated on pruned eggplant seedlings, at a concentration of 10⁸ conidia/mL (deposited at 6000±586 conidia mm^?2). The mortality based on mycosis varied from 18 to 97% after 6 days. Isaria fumosorosea isolate Pf04, B. bassiana isolates Bb06, Bb12, and L. lecanii L14 were found the most effective. Furthermore, five isolates were chosen for concentration–mortality response assays and compared to B. bassiana GHA as a standard. The numbers of nymphs infected by fungi were correlated with the spore concentration. L. lecanii L14 and I. fumosorosea Pf04 had the shortest LT₅₀ at 3.5 and 3.3 days at 6000±586 conidia mm^?2. Mortality declined and LT₅₀ values were longer as the concentration of conidia was reduced. The LD₅₀ values were calculated as 87, 147, 191, 263, and 269 conidia mm^?2 for isolates L14, GHA, AS1260, Bb13, and Pf04, respectively. These results indicated that the Q biotype of sweetpotato whitefly was susceptible to the five isolates of entomopathogenic fungi and these isolates have potential to be developed as microbial pesticides for whitefly control.     

Persistence and efficacy of a Beauveria bassiana biopesticide against the house fly,Musca domestica,on typical structural substrates of poultry houses

Entomopathogenic fungi, such as Beauveria bassiana, offer potential for use as biopesticides for control of house flies in poultry production facilities. This study evaluates persistence and efficacy of oil-formulated B. bassiana conidia against adult house flies on a range of structural substrates commonly found in poultry houses. Exposure of flies to fungal-treated surfaces produced high levels of infection leading up to 100% mortality in 6–10 days. However, the infectivity of the spray residues declined rapidly within 1 or 2 weeks following repeated fly exposures. Investigations showed that, in the absence of flies, conidia remained viable on test surfaces for up to 3 months regardless of substrate type, application method or fungal production batch. Rather, it was the presence of flies themselves that was responsible for reducing persistence. The exact mechanisms remain unclear but involve a combination of physical removal and chemical deactivation, with decay rates increasing at higher fly densities. While the rapid decay could pose a challenge for operational use, the results suggest it might be possible to tailor treatment frequencies to fly densities with, for example, weekly applications at high fly densities and longer intervals when populations decline. Further research is needed to determine persistence in semifield and field settings and to quantify the influence of fly densities under natural exposure conditions.     

Effect of dual inoculation with nematodes and fungal pathogens on the survival of Phyllophaga polyphylla larvae (Coleoptera: Scarabaeidae)

Phyllophaga polyphylla (Coleoptera: Scarabaeidae) is an important pest of maize and other crops in Mexico. Previous reports showed that this pest was highly resistant to fungal and nematode infection when each pathogen was inoculated separately; in this study, we evaluated whether dual inoculation of fungi and nematodes, in all possible pair-wise combinations and orders of inoculation and including an evaluation of a time separation of 73 hours between each pathogen's inoculation, would increase mortality in P. polyphylla larvae. The pathogens were two isolates of Heterorhabditis bacteriophora applied at a concentration of 50 infective juveniles (IJ) mL^–1, an isolate of Beauveria pseudobassiana and of Metarhizium pingshaense both applied at a concentration of 1 × 10⁸ conidia mL^–1. In the first experiment, the combined mortality when pathogens were dual-inoculated (13%), although significantly higher than single-inoculated treatments (8%), demonstrated that antagonistic interactions were ongoing between the pathogens, as confirmed by the χ²-test. In a separate experiment, using only the B. pseudobassiana isolate (1 × 10⁸ conidia mL^–1) and one isolate of H. bacteriophora (100 IJ mL^–1), we studied the effect of different order of inoculations but included a two-week separation between inoculation of each pathogen. Mortalities obtained were similar to the previous experiment; all interactions resulted in antagonistic effects, except when the fungal pathogen was inoculated first, which resulted in an additive interaction. Understanding the mechanisms for the interaction requires further study but, for practical biological control, we suggest that more virulent fungal and nematode isolates are necessary to achieve control of P. polyphylla.     

Culture of Entomophthora muscae in vivo and its infectivity for six species of muscoid flies

A method for the continuous culture of Entomophthora muscae (Entomophthoraceae) in adult house flies (Musca domestica) is described. Using this method we have maintained the host-pathogen system in the laboratory for more than one year. The ability of this isolate to cause fatal infections in virtually all of the house flies at risk has remained constant. The face fly (Musca autumnalis), the onion fly (Hylemya antiqua), and the seed-corn fly (H. platura) are susceptible to this isolate. The stable fly (Stomoxys calcitrans), the black blow fly (Phormia regina), and the false stable fly (Muscina stabulans) are not.     

Evaluation of Beauveria bassiana and B. brongniartii strains and four wild-type fungal species against adults of Bactrocera oleae and Ceratitis capitata

The virulence of two isolates of the hyphomycete fungi, Beauveria bassianaand B. brongniartii, and additional fungal species isolated from diseased Bactrocera oleae pupae and Sesamia nonagrioideslarvae were assessed against adults of the olive fruit fly B. oleae and the Mediterranean fruit fly Ceratitis capitata (Diptera: Tephritidae). Contact and oral bioassays revealed that moderate to high mortality rates for the olive fruit fly occurred when the adults were exposed to conidia of Mucor hiemalis, Penicillium aurantiogriseum, P. chrysogenum and B. bassianaisolates. A strain of M. hiemalis isolated from S. nonagrioides larvae was the most toxic resulting in 85.2% mortality to the olive fruit fly adults. B. brongniartiiand B. bassiana were the most pathogenic to the C. capitataadults causing 97.4 and 85.6% mortality. Metabolites collected from the M. hiemalis and P. chrysogenum isolates were toxic to adults of both species.