Environmental contamination with Metarhizium anisopliae from fungal bands for control of the Asian longhorned beetle, Anoplophora glabripennis (Coleoptera: Cerambycidae)

Our objective was to determine whether conidia from non-woven fiber bands impregnated with Metarhizium anisopliae F 52 could be spread to other parts of the environment, and whether Anoplophora glabripennis could become infected with conidia dispersed from fungal bands. In the field, bands containing M. anisopliae were hung at 3 m height on 15 trees. Bark samples were taken 10, 20, and 30 cm above the bands and 10, 30, and 60 cm below the bands 2, 5, and 9 days after band placement to quantify conidial densities. There were more conidia in samples taken below bands compared with samples taken above bands. A significant positive correlation was found between rainfall and the occurrence of conidia on any of the bark samples. A laboratory study was conducted to determine whether conidia from M. anisopliae fungal bands could be spread to other parts of the environment by A. glabripennis adults and whether A. glabripennis could become infected by conidia dispersed from fungal bands. One or five adult A. glabripennis were used to contaminate artificial environments with conidia. All adults subsequently exposed to contaminated environments were killed by fungal infection. Beetles exposed to environments that had been contaminated by five beetles died in fewer days compared with environments contaminated by one beetle. Beetles in both density treatments died in fewer days than beetles exposed to environments without M. anisopliae conidia. Our results indicate that environmental contamination with conidia from fungal bands can occur, and that adult A. glabripennis can acquire infection from a contaminated environment.     

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.     

Pathogenicity of the fungus, Verticillium lecanii, to the green peach aphid, Myzus persicae (Hom.: Aphididae)

Pathogenicity of the hyphomycete, Verticillium lecanii (DAOM 198499), was investigated to aphid, Myzus persicae under laboratory conditions. Analysis of lethal effect of six various concentrations 0, 10(4), 10(5), 10(6), 10(7) and 10(8) conidia/ml of V. lecanii against third nymphal stage of M. persicae, indicated significant mortality on aphids. Mean comparisons showed that there was significant difference between treatments. Three days after treatment, aphid mortality observed and after 12 days minimal mortality was 17.77 in control and maximum was 100 percent related to 10(7) and 10(8) conidia/ml. LC50 and LT50 values were estimated by probit analysis and life test. LC50 value for aphid mortality was 1.4 x 10(4) conidia/ml, LT50 values for concentrations 10(4), 10(6), 10(6), 10(7) and 10(8) conidla/ml was 10, 10, 9, 8 and 6 days, respectively. In this experiment, net reproduction rate of aphid's (R0) decreased significantly when concentration increased. These observations showed that V. lecanii (DAOM 198499) can be an active biological agent against aphids.     

Aerial treatment of the Australian plague locust, Chortoicetes terminifera (Orthoptera: Acrididae) with Metarhizium anisopliae (Deuteromycotina: Hyphomycetes)

Between October 1999 and April 2000, nearly 4000 ha of nymphal bands and adult swarms of Chortoicetes terminifera (Walker) were aerially treated using a ULV oil formulation of strain FI-985 of Metarhizium anisopliae var. acridum. During the mild weather (maxima 22-30 degrees C) of spring (October), there was little change in nymphal bands during the first week but at all doses between 25-100 g (1-4 x 10(12) conidia) ha(-1), the bands rapidly declined 9-12 days after treatment reaching > 90% mortality by 14 days. Metarhizium persisted for some time as there was 50% mortality of locusts fed vegetation collected from the treated blocks seven days after treatment. Persistence was confirmed by the high mortality of bands that invaded from untreated areas and of nymphs that hatched on the plot five to seven days after treatment, though mortality was then delayed until early in the third week. During summer (January), temperatures were high (maxima 36-42 degrees C), and at all doses between 25 and 125 g (1-5 x 10(12) conidia) ha(-1), there was a rapid decline seven to ten days after treatment. By 12-14 days, there was a > 90% decline in numbers in most blocks which was confirmed by helicopter surveys two weeks after treatment that found very few adults within or near treated areas. Mortality was delayed in the high dose where there were blockages of spray equipment during treatment. The clear demonstration that Metarhizium can suppress small local populations of C. terminifera led to the limited operational use of Metarhizium on an organic farm and in a National Park where nearly 2500 ha of bands and swarms were treated. Continued research is needed to develop a commercially viable product so that Metarhizium can form a significant part of a programme of integrated pest management of locusts in Australia.     

A comparison of the virulence,stability and cell-wall-surface characteristics of three spore types produced by the entomopathogenic fungus Beauveria bassiana

Summary Beauveria bassiana can produce three spore types; aerial conidia, submerged conidia and blastospores. We have examined the spore surface characteristics (hydrophobicity and cell-wall surface lectins), thermal inactivation and the virulence towards the migratory grasshopper, Melanoplus sanguinipes, of each of the three spore types. The hydrophobicities of the aerial and submerged conidia were quite similar. Blastospores were less hydrophobic than either of the two types of conidia. Hydrophobic interactions are thought to play a significant role in attachment of the spore to the host organism. However, the less hydrophobic blastospores were slightly more virulent (LT₅₀ of 6.50 days) when compared to the aerial and submerged conidia (7.12 and 7.24 days), respectively. The lectin-binding characteristics of the aerial and submerged conidia were very similar but differed from that of blastospores. Growth of blastospores on a variety of carbohydrates did not affect their lectin-binding characteristics. Spore viability measurements showed that aerial and submerged conidia retained their viability for a longer period than blastospores. The similarity in hydrophobicity, stability, virulence and lectin-binding of aerial and submerged conidia make the latter an ideal candidate for mycoinsecticide production since they can be recovered after growth on inexpensive substrates.Offprint requests to: G. G. Khachatourians     

Effects of the Sunscreen Oxybenzone on Field Efficacy and Persistence of Metarhizium flavoviride Conidia Against Kraussella amabile (Orthoptera: Acrididae) in Mali,West Africa

A field trial was performed at Mourdiah, northwest Mali, to evaluate whether the performance of Metarhizium flavoviride conidia would improve if 2% oxybenzone, an oil-soluble ultraviolet light protectant, was used. Conidia were applied in an ultra-low volume formulation against nymphs of the grasshopper Kraussella amabile . Mortality was assessed by collecting grasshoppers directly from field plots (sweep net samples) or by exposing untreated nymphs to sprayed vegetation for 48 h (field cage samples). For sweep net samples, the mortalities in M. flavoviride treatments varied between 94.5 and 99.9% up to 7 days after treatment, while control mortalities were significantly lower (20.7-55.0%). There were no significant differences between M. flavoviride treatments with or without oxybenzone. The average survival times were found to be significantly lower for fungal treatments (4.9 days) compared with the control treatment (11.8 days). W ith field cage samples, the mortality in M. flavoviride treatments declined from > 95.0% for nymphs exposed just after application to < 30.0% for nymphs exposed to treated vegetation on the 10th day after application. The control mortalities varied between 5.1 and 37.2%, and were significantly lower than fungal treatments for samples placed just after application and 10 days after application. The overall average survival times were 10.9 and 10.7 days for M. flavoviride with and without oxybenzone respectively, and 13.9 days with the control treatment. An exponential regression was fitted using cage mortality data which indicated that the half-life for conidial infectivity was 4.3 days on treated vegetation.     

Effects of Metarhizium anisopliae on Meccus pallidipennis (Hemiptera: Reduviidae) over different types of wall surfaces

The entomopathogenic fungus Metarhizium anisopliae is virulent for the insect triatomine Meccus pallidipennis. To evaluate the functionality of a fungal formulation (vegetable oil and emulsifiers) of this fungus, virulence was assayed by insect mortality on the pronotum of third instar nymphs (N3) M. pallidipennis in laboratory conditions and ST₅₀ was calculated. Mortality was evaluated directly: 100%, 97.33% and 98.66% mortalities were caused by formulation, emulsified formulation and fungal conidia, respectively, at day 8 of insect infection. Another bioassay was carried out in simulated external conditions (peridomicility) using red and gray brick walls, a stone fence and mountain soil (experimental units). These simulated conditions were infected with 10?ml of a 1?×?10⁹?conidia/ml emulsified formulation by means of a manual sprinkler prior to the placement of the nymphs. Ten N3 M. pallidipennis were deposited in each experimental unit and insect mortality was monitored every 12?h for 22 days. Each treatment was replicated four times. With the red brick wall, a mortality of 90% at day 22 and a ST₅₀ of 15 days were obtained on N3 M. pallidipennis; with the gray brick wall, 100% mortality and a ST₅₀ of 13 days; and with the stone fence, 88.33% mortality and a ST₅₀ of 21 days. The results obtained in this research work indicate that the formulation with conidia of the M. anisopliae strain EH-473/4 may be auxiliary in the development of strategies for the control of Chagas disease insect transmitters such as M. pallidipennis.     

Potential of a Dried Rice/Mycelium Formulation of Entomopathogenic Fungi to Suppress Subterranean Pests in Small Fruits

The potential of a dried rice/mycelium formulation of three species of entomopathogenic fungi ( Metarhizium anisopliae, Beauveria bassiana and Paecilomyces farinosus ) was assessed according to five criteria: storage life, field efficacy against two subterranean pests of cranberry, persistence and efficacy against first-instar black vine weevil larvae in potted strawberry and field persistence in a cranberry bog. The number of conidia sporulated from the formulation did not decline between 4 and 20 days. In small plot field trials, the numbers of black vine weevil, Otiorhynchus sulcatus (F.), were significantly lower in plots treated with 1000 gm - 2 of formulated M. anisopliae than in untreated plots. In similar trials, the numbers of adult cranberry girdler, Chrysoteuchia topiaria (Zeller), inside emergence cages were significantly lower on plots treated with the formulated M. anisopliae than on untreated plots. The formulation suppressed first-instar black vine weevil larvae in potted strawberry regardless of the species of entomopathogenic fungus or the post-inoculation interval. M. anisopliae was periodically isolated for up to 637 days from plots in cranberry bogs treated with the formulation. In general, the dried rice/mycelium formulation may be a useful fungal mycopesticide for the management of subterranean pests of small fruits, especially if applied at high rates to areas of serious pest infestation.     

Field observations of the effects of fenitrothion and Metarhizium anisopliae var. acridum on non-target ground dwelling arthropods in the Sahel

The effect of the chemical insecticide, fenitrothion, and a mycoinsecticide based on Metarhizium anisopliae var. acridum on the activity of non-target epigeal arthropod scavengers was investigated in areas of open savannah in southeast Niger Republic, West Africa. Both insecticides were applied as full cover sprays to unreplicated 800 ha plots to assess their season-long control of Sahelian grasshoppers. Compared with control plots, fenitrothion caused an immediate but temporary reduction in grasshopper numbers, whereas M. anisopliae var. acridum provided delayed but prolonged control. Scavenging rates of pyrethroid-killed grasshoppers placed along transects in unsprayed plots and those treated with fenitrothion and M. anisopliae var. acridum at various intervals after spraying were assessed. In the fenitrothion plot, an immediate reduction in scavenging activity occurred that was still apparent after 40 days at the plot center, although recovery at the plot edges was more rapid. By contrast scavenging rates remained high over equivalent areas in the M. anisopliae var. acridum and two untreated plots. Concurrent to the scavenging study, counts of grasshopper cadavers resulting from the spray treatments were conducted. These counts revealed that the density of grasshopper cadavers remained low throughout the M. anisopliae var. acridum plot and explained <1% of the reduction in live grasshoppers resulting from treatment, compared with >20% in the fenitrothion plot. This shortfall in grasshopper cadavers resulting from the spray treatment in the M. anisopliae var. acridum plot was unexpected because in a monitoring study, fungus-killed (unlike pyrethroid-killed) grasshoppers were unattractive to scavengers and readily persisted in this plot, and thus should have become apparent. Given we did not observe significant grasshopper dispersal, the scarcity of cadavers generated in the M. anisopliae var. acridum plot, together with unquantified visual observations, suggests that predation of infected but living grasshoppers was high. Our data provide circumstantial evidence that the different effects of chemical and biological grasshopper control on grasshopper natural enemies may influence the efficacy of large-scale treatments.     

Experimental studies to evaluate spray applications of a mycoinsecticide against the rice grasshopper, Hieroglyphus daganensis, in northern Benin

Three experiments were conducted to examine the effects of spray applications of a mycoinsecticide against the rice grasshopper, Hieroglyphus daganensis, in northern Benin. The experiments were designed to expand on previous evaluation studies and contribute to a better understanding of the overall impacts of mycoinsecticide applications. The first experiment showed that acquisition of spores from the spray residue had a significant effect on mortality rate and a greater single influence on mortality than direct contact with spores from the spray aerosol. The second and third experiments revealed the mycoinsecticide to have sub- and pre-lethal effects, causing a significant reduction in per capita feeding rate and increasing susceptibility to predation. The significance of these results for the overall impact of spray applications and for development of comprehensive evaluation methods are discussed.     

Effect of Bait Substrate and Formulation on Infection of Grasshopper Nymphs by Beauveria bassiana

The effects of two formulations (oil and water) and two bait substrates (lettuce and bran) on infection of grasshopper nymphs (Melanoplus sanguinipes) by Beauveria bassiana were investigated. More nymphs died of mycosis after they had ingested substrates inoculated with conidia in oil than in water, but there was no difference between the lettuce and bran substrates. Of the conidia recovered in frass, most (95%) were recovered within 24 h of ingestion of the lettuce and bran substrates by nymphs. Significantly more conidia averaged over time were recovered in frass from the water than from the oil formulation. A higher incidence (33-82%) and more rapid onset of mycosis was observed in nymphs that were surface-sterilized before ingestion, compared with those surface-sterilized after ingestion of lettuce and bran substrates inoculated with B. bassiana in both formulations. A similar trend was observed in nymphs receiving the sterilization treatment before, rather than after, ingestion of wheat leaves sprayed with conidia in oil or water. Numerous conidia were observed on the heads, thoraxes and abdomens of nymphs that ingested treated lettuce or bran. Modest numbers of conidia were also recovered from the surfaces of nymphs, but no differences were observed between formulations or substrates. However, when nymphs ingested lettuce disks treated with fluorescent dye in either oil or water, more dye was observed on nymphs with the oil formulation. This laboratory study demonstrates that grasshopper nymphs are highly susceptible to infection by conidia formulated on bait substrates, and that the efficacy of the bait relies on the extent to which nymphs become surface-contaminated during ingestion.     

黄绿绿僵菌Mf82悬乳剂对褐飞虱作用的时间-剂量-死亡率模型分析

为了寻找褐飞虱Nilaparvata lugens生物防治的新途径,用新分离出的黄绿绿僵菌Metarhiziumflavoviride(Mf82)菌株与实验室保存的黄绿绿僵菌、金龟子绿僵菌和球孢白僵菌3种菌种9个菌株作对比,测定了它们对褐飞虱成虫的毒力。结果表明:Mf82菌株对褐飞虱成虫的毒力最高,以1.0×108个孢子/mL的孢子液喷雾接种到褐飞虱成虫体表上,累积死亡率高达81.7%,LT50为4.6d,致病效果显著高于其他受测菌株。在此基础上研制了黄绿绿僵菌悬乳剂,并研究了其对褐飞虱的致病力。结果表明:随着黄绿绿僵菌浓度的增加,褐飞虱的累计死亡率增加,在浓度为1,048个孢子/mm2时,累计死亡率达到85.0%。利用时间-剂量-死亡率模型对数据进行处理,所建模型均顺利通过Hosmer-Lemeshow拟合异质性检验,表明模型拟合良好,并由模型估计出了该剂型对褐飞虱的致死剂量与致死时间。在接种后第7天和第9天,LC50值分别为2.1×103、9.9×102个孢子/mm2,LC90分别为7.8×104、3.7×104个孢子/mm2。黄绿绿僵菌悬乳剂对褐飞虱的致死时间与对数剂量相关,供试菌剂LT50值随着对数剂量的增加而递减,对数剂量由7.0增加到8.0时,LT50由8.9d降为5.7d。可见该黄绿绿僵菌悬乳剂对褐飞虱具有较强的毒力,在褐飞虱生物防治中具有广阔的应用前景。     

Protection of entomopathogenic conidia against chemical fungicides afforded by an oil-based formulation

We evaluated the protection afforded by an oil formulation against non-compatible fungicides in mixtures with conidia of the entomopathogenic fungi Metarhizium anisopliae (Ma) and Beauveria bassiana (Bb). Under laboratory conditions, viability of unformulated (aqueous suspensions) Ma conidia was harmed by recommended label doses of carbendazim (not tested for Bb), and both Ma and Bb conidia were affected by triadimefon. On the other hand, effect of fungicides was usually nil or minimal on conidia formulated as oil-containing suspensions (emulsifiable oil + water). Germination rates for unformulated and oil-formulated Ma conidia subjected to carbendazim were reduced by 77.3 and 12.1%, respectively, compared to their fungicide-free counterparts. Germination rates at 16 h post-inoculation for unformulated and oil-formulated Bb conidia subjected to triadimefon were reduced by 20.5 and 5.5%, respectively, compared to their fungicide-free counterparts. No differences were observed at 20 h post inoculation, indicating a fungistatic action of this compound on Bb conidia. Virulence of unformulated conidia amended with fungicides against third instar Diatraea saccharalis larvae was negatively affected compared to their formulated counterparts. These results suggest that oil-formulated conidia can be effectively protected from damage caused by chemicals, which could have applications in tank mixing or alternate applications with shared spraying equipment, being especially relevant for IPM programs in which mycopesticides and chemicals are simultaneously sprayed.     

Susceptibility of Ixodes scapularis (Acari: Ixodidae) to Metarhizium brunneum F52 (Hypocreales: Clavicipitaceae) using three exposure assays in the laboratory

An emulsifiable concentrate (EC) and granular (G) formulation of the entomopathogenic fungus, Metarhizium brunneum strain F52 (formerly Metarhizium anisopliae strain F52) were tested against unfed adults and nymphs of Ixodes scapularis Say in the laboratory. Three exposure methods; dip, surface contact, and direct spray application, and three exposure time intervals (3, 30, and 300 min) were used to evaluate the EC formulation. Application rates ranged from 2.6 x 10(2) to 2.6 x 10(8) conidia/cm2. The surface treatment was used for granular formulation with concentrations ranging from 2.3 x 10(5) to 2.3 x 10(7) conidia/cm2 for same three exposure times. Both the EC and G formulations of this fungus were highly pathogenic against I. scapularis adults and nymphs. Logistic regression analysis found formulation, spore concentration, time of exposure, and observation period were significant or highly significant factors influencing tick mortality. For adult I. scapularis, the spray application with the EC formulation of M. brunneum F52 resulted in a lower LC50 (5.9 x 10(4) conidia/cm2) at 30 min than surface exposure to the EC (LC50 = 1.3 x 10(6) conidia/cm2) or G formulation (LC50 = 8.1 x 10(5) conidia/cm2). At higher concentrations, fungal activity was evident in adult I. scapularis held at 5 degrees C suggesting the fungus may provide control in the cooler fall season. While the observed pathogenicity of a fungus against ticks can be dependent upon the bioassay assessment, we found nymphs and adults of I. scapularis to be highly susceptible to M. brunneum F52, regardless of the exposure method used.     

Field Treatment of Desert Locust ( Schistocerca gregaria Forskal) Hoppers in Mauritania Using an Oil Formulation of the Entomopathogenic Fungus Metarhizium flavoviride

A field trial was conducted to test the efficacy of an oil formulation of the entomopathogenic fungus , Metarhizium flavoviride Gams & Rozsypal (Deuteromycotina: Hyphomycetes) , as a control agent for the desert locust , Schistocerca gregaria Forskal (Orthoptera: Acrididae) , in Mauritania . The treatment was applied to hopper bands in the field . In caged samples of the treated insects , 99% mortality was observed in 15 days . On average during application only 51% of the hoppers received spray droplets directly , the additional mortality being due to pick - up of spores from the spray residue . This mortality was assessed by exposing insects to a treated plot at intervals , and monitoring disease levels during subsequent incubation in cages . The spray residue remained highly infective for a period of 6 days after application . The number of nymphs in the hopper bands was estimated using a computer - based photographic image - processing technique . Unfortunately , most of the hopper bands dispersed into small groups , but a population reduction could be demonstrated for one hopper band . Maximal daily mortality in the field was observed 10 - 11 days after application , compared with 6 - 10 days in cages .     

A Low-cost Contamination Device for Infecting Adult Tsetse Flies, Glossina spp., with the Entomopathogenic Fungus Metarhizium anisopliae in the Field

A low-cost device for infecting adult tsetse fly, Glossina fuscipes fuscipes , with the entomopathogenic fungus Metarhizium anisopliae was designed and tested in the field. Tsetse flies that are attracted to the trap entered the contamination device and ultimately became infected with the fungus. Traps exposed to the sun attracted more flies than did the ones placed in the shade. The time spent by single flies in the contamination device varied between 5- 189 s, and the subsequent number of conidia collected varied between 1.6 ×10 5 conidia and 40.5 ×10 5 conidia per fly, and largely depended on the behavior of individual flies. Dry conidia of M. anisopliae in the device retained their viability for 31 days in the field, and efficacy against G. fuscipes was not affected.     

FI-1045: A Profile of a Commercially Useful Isolate of Metarhizium anisopliae var. anisopliae

The isolate FI-1045 is the basis of a mycoinsecticide, BioCane TM granules recently registered for the control of greyback canegrub, Dermolepida albohirtum (Coleoptera: Scarabaeidae: Melolonthinae) in Australian sugarcane fields. The isolate was obtained from a naturally infected larva of D. albohirtum collected from Tully in north Queensland. The isolate can be distinguished from others infecting the same insect and also other species of canegrub by means of RAPD patterns and sequence data from the ITS region. A comparison of a stored FI-1045 isolate with three derived isolates which had different histories of host-passage, showed no variation in RAPD pattern. All isolates grew well at temperatures between 20 and 30°C but did not grow at 35°C and grew slowly at 15°C. However, on potato dextrose agar, the original FI-1045 grew more rapidly and did not produce as much pigment as the derived isolates. It is speculated that this difference was due to the storage method used with the original FI-1045 being stored at -70°C and the other isolates being freeze-dried. Bioassays against third instar greyback canegrubs gave a mean LC 50 of 8.7 ×10 4 conidia g -1 peat substrate after 10 weeks. Using Tenebrio molitor as a host, it was found that conidia taken directly from the infected insect were similar in virulence to the cultured FI-1045. Using injection of culture filtrate as the assay method, it was found that FI-1045 produced destruxins. In laboratory host range tests, a dose of 10 6 conidia g -1 peat killed 96% of southern oneyear canegrubs, Antitrogus consanguineius , 85% of Lepidiota picticollis and less than 30% of the other five species of canegrub tested.     

Identification of Metarhizium strains highly efficacious against Aedes,Anopheles and Culex larvae

Entomopathogenic fungi, such as Metarhizium anisopliae and Beauveria bassiana, have been shown to be efficacious in killing mosquito larvae of different mosquito species. The current study compared the pathogenicity and efficacy of two formulations of three fungal strains against different instars of three mosquito species with the aim of identifying the most virulent strain for use under field conditions. Three strains of Metarhizium, ARESF 4556, ARSEF 3297 and V275, were assayed against early (L_2?3) and late (L_3–4) instar larvae of Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. Two formulations of the fungi were tested, dry conidia and aqueous suspensions (i.e. ‘wet’ conidia). Effects of all combinations of conidia, mosquito species, instar, fungal strain and concentration on mosquito mortality were analysed using Cox regression and Kaplan–Meier analyses. Strain ARSEF 4556 was more virulent than ARSEF 3297 and V275, with LT₅₀ values ranging from 0.3 to 1.1 days, with Anopheles and Culex being more susceptible than Aedes. Early and late instars were equally susceptible independent of species. Although the formulation did influence mortality rates, both ‘wet’ and ‘dry’ conidia applications were highly effective in killing mosquito larvae. Viable spores were more efficacious than heat killed spores. The latter did cause mortality but only at high concentrations. Metarhizium sp. has proved to be effective in reducing survivability of all larval stages of Aedes, Anopheles and Culex under laboratory conditions. Aedes larvae were generally more tolerant than Anopheles and Culex irrespective of fungal strain.     

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.     

Selection of an Isolate of the Insect Pathogenic Fungus Metarhizium anisopliae Virulent to the Lettuce Root Aphid, Pemphigus bursarius

The virulence of hyphomycete entomopathogenic fungi was measured in laboratory bioassays against the lettuce root aphid, Pemphigus bursarius, a serious pest of field lettuce grown in the UK. Of 25 isolates of fungi examined, only one isolate, Metarhizium anisopliae 391.93, killed lettuce root aphids consistently. This fungus was isolated originally from the closely related saltmarsh aphid, P. trehernei. The median lethal concentration of conidia at 10 days post6- 1 inoculation estimated from five independent bioassays was 2.45 × 10⁶ conidia ml^-1. The fungus had no significant effect on the mean number of offspring/aphid produced, but it sporulated 6 profusely on host cadavers, producing approximately 4 × 10⁶ conidia/cadaver 14 days after treatment, and diseased aphids died attached to plant roots. It thus has the potential to spread through densely packed colonies of P. bursarius feeding on the roots of susceptible or partially resistant plants.