Development of a novel bioassay for estimation of median lethal concentrations (LC50) and doses (LD50) of the entomopathogenic fungus Beauveria bassiana, against western flower thrips, Frankliniella occidentalis

To conduct laboratory experiments aimed at quantifying secondary acquisition of fungal conidia by western flower thrips (Frankliniella occidentalis), an efficient assay technique using Beauveria bassiana as the model fungus was developed. Various application protocols were tested and it was determined that the percent mortality did not vary among protocols. Peak mortality of second-instar nymphs, under constant exposure to conidia, occurred 5 days post-inoculation. Second-instar thrips that were exposed to conidia within 24 h of the molt to second instar were more susceptible to Beauveria bassiana than thrips exposed after times greater than 24 h post-molt. Conidia efficacy, which was monitored at 24 h intervals, did not differ significantly within 72 h. A test of the final bioassay system was conducted in a series of assays aimed at determining the LD50 of B. bassiana technical powder against second-instar western flower thrips. It was determined that B. bassiana (strain GHA) is highly effective at very low doses (LD50 of 33-66 conidia/insect).     

Antagonism between Beauveria bassiana and imidacloprid when combined for Bemisia argentifolii (Homoptera: Aleyrodidae) control

Imidacloprid and the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin are both used to control the whitefly Bemisia argentifolii Bellows & Perring. We tested whether the two control strategies acted additively, synergistically, or antagonistically when combined for whitefly control. We found antagonism in that B. bassiana inhibited the effectiveness of imidacloprid. When B. bassiana was combined with imidacloprid, insect response was either less than or similar to (depending on B. bassiana rates) that when imidacloprid was used alone. Adding imidacloprid to B. bassiana treatments always increased mortality, but the increase was less than additive. Beauveria bassiana spore germination and colony formation were not inhibited by imidacloprid in vitro, and B. bassiana did not adsorb or degrade imidacloprid in a tank mix. We hypothesize that B. bassiana caused a behavioral response that reduced insect feeding and uptake of imidacloprid.     

Trials of Beauveria bassiana, Paecilomyces fumosoroseus and imidacloprid for management of Trialeurodes vaporariorum (Homoptera: Aleyrodidae) on greenhouse grown lettuce

Mineral oil-based emulsifiable preparations of Beauveria bassiana (Bb) and Paecilomyces fumosoroseus (Pfr) conidia were separately applied alone or together with low rates of imidacloprid 10% WP at 4.7% (Im 1), 14.0% (Im 2), and 23.3% (Im 3) of its recommended application rate, respectively, against the greenhouse whitefly, Trialeurodes vaporariorum, on lettuce grown in the greenhouse. Besides eight fungal treatments, the three low application rates of imidacloprid in the oil-based carrier and a blank control (CK) were also included as treatments of the trials conducted in 2002 and 2003. For the 12 treatments of each trial with three replicates, 1,000-fold aqueous dilutions were sprayed twice on transplanted lettuce at a 15-day interval at a rate of ∼1.43×10¹³ conidia ha^-1 for each fungal treatment or at one of the low rates of imidacloprid using a backpack hand-operated hydraulic sprayer. Based on whitefly densities, mortalities, relative efficacies and percent density declines estimated from whitefly counts made at 5-day intervals, all B. bassiana and P. fumosoroseus sprays were highly effective against T. vaporariorum compared to CK and Im 1-3 in both trials. In trial 1, the estimates of whitefly density decline and relative efficacy ranged from 44 and 72% (Bb) to 79 and 90% (Pfr+Im 2-3) on day 10 after the first spray and exceeded 94% for all fungal treatments 10 days after the second spray. Similar trends in whitefly control were also achieved in the corresponding treatments of trial 2. A more desirable and faster control resulted from fungal sprays containing more imidacloprid, but none of the three low imidacoprid rates alone suppressed the whitefly population more effectively than any fungal treatment despite a varying degree of efficacy. Moreover, P. fumosoroseus tended to be more effective against T. vaporariorum than B. bassiana when applied alone or together with the same low rate of imidacloprid but the difference in whitefly control eventually achieved was not significant between the two fungal agents. Thus, the emulsifiable preparations of both fungal agents can be considered as alternatives for whitefly management and for a slightly faster control can be applied together with imidacloprid at around 15% of its common application rate. An economic balance between the efficacy of whitefly control and the ease and cost of conidial production is needed when both fungi are incorporated into whitefly management.     

Field-Cage Studies of Beauveria bassiana (Hyphomycetes: Moniliaceae) for the Suppression of Adult Western Corn Rootworm, Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)

The efficacy of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin, was tested as a control agent for adult western corn rootworm, Diabrotica virgifera virgifera LeConte, in walk-in field cages. Suspensions of B. bassiana conidia were applied to corn plants in cages into which laboratory-reared beetles had been released. Beetles were collected at 3 and 5 days post-application and evaluated in the laboratory for mortality. Mortality was 10, 29 and 50%, at rate equivalents of 7 X 10 12 , 2 X 10 13 (two applications), and 5 X 10 13 conidia/ ha, respectively. There was no significant difference in mortality of beetles collected at 3 days compared with 5 days post-application. Mortality due to B. bassiana was 24% when beetles were released into field cages 24 h post-application (5 X 10 13 conidia/ha) compared with 50% when beetles were present during the application. Beetle mortality declined significantly with increasing time from application in feeding assays carried out with leaf samples removed from plants at 0, 12, 24 and 72 h post-application. Mortality of beetles collected from treated plants within cages and maintained in the laboratory was found to overestimate the population decline by 10% when compared with beetle estimates from treated plants within field cages.     

Insecticidal properties of a Chenopodium-based botanical

The emulsifiable concentrate UDA-245 based on an essential oil extract from Chenopodium ambrosioides variety near ambrosioides, a North American herbaceous plant, was compared with commercially available pesticides for their effectiveness to control green peach aphid, Myzus persicae (Sulzer) (Homoptera: Aphididae), western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), and greenhouse whitefly, Trialeurodes vaporariorium (Westwood) (Homoptera: Aleyrodidae). Side effects on the whitefly parasitoid Encarsia formosa Gahan (Hymenoptera: Aphelinidae) also were determined. With green peach aphid, UDA-245 at 0.5% concentration was significantly more effective than the control (water) treatment in a laboratory bioassay and significantly more effective than neem oil and the control treatment and as effective as insecticidal soap in a greenhouse assay. With the western flower thrips, UDA-245 at 0.5% was significantly more effective than neem oil, insecticidal soap and the control treatment in a laboratory bioassay, whereas in a greenhouse assay, UDA-245 at 1.0% was the only treatment that maintained control of the western flower thrips 2 wk after the last treatment period. UDA-245 at 0.5% (laboratory bioassay) was significantly more effective in managing greenhouse whitefly than neem oil, endosulfan, and the control treatment and as effective as insecticidal soap. Insecticidal soap proved to be toxic to the parasitoid E. formosa (71.9% mortality), whereas UDA-245 at 0.5% was not significantly more toxic than the control (11.2 and 4.6% mortality, respectively). Our results suggest that a greenhouse integrated pest management (IPM) program using a botanical such as UDA-245 could effectively control infestations of major pests present while having a negligible effect on biological control agents.     

Evaluation of synergistic interactions between the Colorado potato beetle (Coleoptera: Chrysomelidae) pathogen Beauveria bassiana and the insecticides, imidacloprid, and cyromazine

Laboratory studies investigated the interaction between the fungal entomopathogen Beauveria bassiana (Balsamo) Vuillemin and sublethal doses of the insecticides imidacloprid and cyromazine when applied to larvae of the Colorado potato beetle, Leptinotarsa decemlineata (Say). When second instars were fed potato leaf discs treated with sublethal doses of imidacloprid and a range of doses of B. bassiana, a synergistic action was demonstrated. Similar results were observed when larvae were sprayed directly with B. bassiana conidia and immediately fed leaf discs treated with imidacloprid. No synergistic interaction was detected when larvae were fed leaf discs treated with sublethal doses ofimidacloprid 24 h after application of B. bassiana conidia to larvae. However, a synergistic interaction was detected when larvae were fed leaf discs treated with imidacloprid and sprayed with B. bassiana conidia 24 h later. Although sublethal doses of both imidacloprid and the triazine insect growth regulator (IGR) cyromazine prolonged the duration of the second instar, only imidacloprid interacted with B. bassiana to produce a synergistic response in larval mortality. In leaf consumption studies, the highest dose of B. bassiana tested promoted feeding in inoculated second instars. Feeding was inhibited when larvae were fed foliage treated with sublethal doses of imidacloprid and significantly reduced when fed foliage treated with a sublethal dose of cyromazine. Starvation of larvae for 24 h immediately after B. bassiana treatment produced a similar result to the combined treatment of B. bassiana and imidacloprid and increased the level of mycosis when compared with B. bassiana controls. Imidacloprid treatment affected neither the rate of germination of B. bassiana conidia on the insect cuticle nor the rate at which conidia were removed from the integument after application. The statistical analysis used to detect synergism and the possible role of starvation-induced stress factors underlying the observed synergistic interactions are discussed.     

Laboratory and field evaluation of Beauveria bassiana against sugarcane stalkborers (Lepidoptera: Pyralidae) in the lower Rio Grande Valley of Texas

Of approximately 18,200 ha planted to sugarcane in south Texas, only approximately 80 ha (<0.5%) are treated with insecticides because this type of control is widely regarded as ineffective against stalkboring pyralids, the key pests of sugarcane. Therefore, nonchemical control measures, such as resistant varieties and biological controls, must be evaluated to mitigate the losses caused by stalkborers. We performed laboratory and field evaluations on the use of Beauveria bassiana (Balsamo) Vuillemin (Deuteromycotina: Hyphomycetes) (strain GHA) against the Mexican rice borer, Eoreuma loftini (Dyar) (Lepidoptera: Pyralidae), which causes damage in south Texas estimated at between 10 and 20 million dollars annually. We also performed bioassays against the sugarcane borer, Diatraea saccharalis (F.) (Pyralidae), the key pest in other sugarcane growing areas. In the bioassays, E. loftini was substantially more susceptible to B. bassiana than D. saccharalis, based on both 5-d LD50 values and survival times. A commercial oil-based formulation of B. bassiana was evaluated in the field using the following treatments: oil alone (control), B. bassiana + oil, and B. bassiana + Silwet L-77 carrier at an application rate of 5 x 10(13) spores per hectare. Neither numbers of E. loftini per stalk, nor stalk damage (approximately 20% bored internodes) were significantly affected by treatment. The application of B. bassiana + Silwet significantly affected the numbers of internodes showing high damage, but not those with low or medium damage. Analysis of yield data and juice quality showed no significant treatment effects. We conclude that the application of Beauveria + Silwet offers the best chances for reducing damage caused by E. loftini of those treatments tested. However, reductions in insect incidence or damage did not result in measurable increases in yield or sugar quality, probably because of insufficient coverage. Effective control of stalkboring pyralids in sugarcane using B. bassiana will likely require improvements in delivery technology.     

Laboratory and greenhouse evaluation of a new entomopathogenic strain of Beauveria bassiana for control of the onion thrips Thrips tabaci

Abstract

The onion thrips Thrips tabaci is one of the most important pests of greenhouse and open-field broccoli, onion and other crops. However, the current strategy of using synthetic pesticides for its control is inadequate and unsustainable, leading to a growing interest in novel and effective biological control alternatives such as entomopathogenic fungi. Among 20 isolates of Beauveria bassiana tested for virulence against T. tabaci in laboratory bioassays, we found strain SZ-26 as the most potent, causing 83–100% mortality in adults at 1×10⁷ mL^?1conidia after 4–7 days. Further experiments in greenhouses showed the strain SZ-26 significantly lowered the numbers of adult and larval stages.     

Biocontrol of almond bark beetle (Scolytus amygdali Geurin-Meneville, Coleoptera: Scolytidae) using Beauveria bassiana (Bals.) Vuill. (Deuteromycotina: Hyphomycetes)

AIMS: To formulate the entomopathogenic fungus Beauveria bassiana in invert emulsion, then apply it against adults of almond bark beetle (Scolytus amygdali) under laboratory and field conditions. METHODS AND RESULTS: The effect of formulated B. bassiana in invert emulsion against S. amygdali adults was shown by comparing the mortality percentage of adults exposed to the formulated fungus using a Petri dish treatment method and by field applications to infested peach trees with mortality of adults exposed to the unformulated fungus or the untreated control. Results obtained from both exposure methods have indicated that treatment of S. amygdali adults with the formulated fungus resulted in a significantly higher mean mortality percentage (P < 0.05) when compared with the treatment with the unformulated fungus or the untreated control. This mortality ranged from 81.2 to 100%, 10 days after treatment with the formulated fungus when compared with 6.7 to 49.6% mortality, 10 days after treatment with the control or the unformulated fungus, respectively. Viability of the fungus conidia in invert emulsion was assessed by calculating the germination percentage of the conidia over time. Results indicated a high storage stability shown by a small loss of germination percentage for the formulated conidia of both strains (5.8 to 8.4% over a 12-week period) vs a low storage stability shown by a high loss of germination percentage for the unformulated conidia of the same strains (58.9 to 61.0% over the same period). The presence of B. bassiana in the galleries of beetles following the treatment of infested trees was shown in the present research. CONCLUSIONS: The results obtained have demonstrated a significantly higher level of efficacy of formulated B. bassiana in invert emulsion against S. amygdali adults under laboratory and field conditions. The ingredients of invert emulsion used in the formulation of the fungus had a negligible effect on the viability of formulated conidia when compared with the unformulated. SIGNIFICANCE AND IMPACT OF THE STUDY: Results obtained in the present research are promising and may be exploited commercially to control S. amygdali adults on various species of stone fruit trees, especially peach trees. This type of biocontrol of this insect may be used as an alternative means to chemical control for management of the insect. No adverse environmental impacts of the fungus or its formulation have been observed during application.     

Effectiveness of attract-and-kill systems using methyl eugenol incorporated with neonicotinoid insecticides against the oriental fruit fly (Diptera: Tephritidae)

Laboratory bioassays and field trials were conducted to evaluate an "attract-and-kill" system using methyl eugenol (ME) with neonicotinoid insecticides against male oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae). In laboratory bioassays, mortality of male flies resulting from the conventional toxicant, naled was 98.3-100% at 24 through 72 h after treatment, whereas the neonicotinoid insecticides imidacloprid and acetamiprid caused only approximately 60-80% at 24 through 72 h after treatment. In the assays of residual effect, naled was persistent up to 96 wk, whereas imidacloprid or acetamiprid was persistent up to 150 wk, resulting in 38.9 or 61.2% male mortality, respectively. Imidacloprid, in particular, caused a delayed lethal effect on flies. In another experiment, male mortality within 28 wk from clothianidin, another neonicotinoid insecticide, was approximately 80% after exposure for 24 h, suggesting a delayed lethal effect similar to those treated with imidacloprid, and mortality was up to 91.8%, if observed, 72 h after treatment. In field trials, attractiveness was similar between ME alone and ME incorporated with naled or neonicotinoids, indicating that addition of these insecticides to ME in traps is not repellent to B. dorsalis males. Using an improved wick-typed trap with longer attractiveness for simulating field application, addition of imidacloprid or acetamiprid maintained 40.1 or 64.3% male mortality, respectively, when assayed once every 2 wk from traps placed in orchards for 42 wk without changing the poison, whereas incorporation with naled resulted in as high as 98.1% after 34 wk and approximately 80% at 42 wk, indicating that persistence is increased compared with sugarcane fiberboard blocks for carrying poison attractants. This study also suggests that neonicotinoid insecticides could be used as an alternative for broad-spectrum insecticides as toxicants in fly traps.     

Laboratory and field evaluations of imidacloprid against Melanoplus sanguinipes (Orthoptera: Acrididae) on small grains

The toxicity of imidaloprid to the migratory grasshopper, Melanoplus sanguinipes (F.), was measured in bioassays, greenhouse trials, and field trials. An LD50 of 53 and 86 ppm for the oral/topical applications of imidacloprid confirmed a low toxicity for this chemical when compared with carbofuran as a standard. However, 100% debilitation was observed at concentrations of > or = 1 ppm. Grasshoppers exhibited leg flexing, abdominal quivering, and tremors before becoming motionless and appearing dead. Knockdown was temporary with a high percentage of recovery within 1 h. Efficacy and feeding damage were determined from artificial infestations of M. sanguinipes at the 2nd, 4th, and early tillering growth stages of winter and spring wheat treated with foliar and seed treatments of imidacloprid. All rates of imidacloprid tested resulted in < 45% mortality to 4th instar and adult M. sanguinipes in the greenhouse and field. Although efficacy was low, high rates of debilitation in bioassays suggest that improved control may be gained by combining imidacloprid with insect pathogens or additional chemicals.     

Relationship between onion thrips (Thrips tabaci) and Stemphylium vesicarium in the development of Stemphylium leaf blight in onion

Stemphylium leaf blight caused by Stemphylium vesicarium and onion thrips (Thrips tabaci) are two common causes of leaf damage in onion production. Onion thrips is known to interact synergistically with pathogens to exacerbate plant disease. However, the potential relationship between onion thrips and Stemphylium leaf blight is unknown. In a series of controlled laboratory and field trials, the relationship between thrips feeding and movement on the development and severity of Stemphylium leaf blight were examined. In laboratory assays, onions (“Avalon” and “Ailsa Craig”) with varying levels of thrips feeding damage were inoculated with S. vesicarium. Pathogen colonisation and leaf dieback were measured after 2 weeks. In pathogen transfer assays, thrips were exposed to S. vesicarium conidia, transferred to onion and leaf disease development was monitored. In field trials, insecticide use was examined as a potential indirect means to reduce Stemphylium leaf blight disease and pathogen colonisation by reducing thrips damage. Results from laboratory trials revealed that a reduction in thrips feeding decreased S. vesicarium colonisation of onion leaves by 2.3–2.9 times, and decreased leaf dieback by 40–50%. Additionally, onion thrips were capable of transferring S. vesicarium conidia to onion plants (albeit at a low frequency of 2–14% of plants inoculated). In field trials, the symptoms and colonisation of Stemphylium leaf blight were reduced by 27 and 17%, respectively with the use of insecticide to control thrips. These results suggest that onion thrips may play a significant role in the development of Stemphylium leaf blight, and thrips control may reduce disease in commercial onion fields.     

Time and concentration dependent interactions of Beauveria bassiana with sublethal rates of imidacloprid against the aphid pests Macrosiphoniellasanborni and Myzus persicae

A series of laboratory bioassays with each consisting of low, medium and high concentration treatments of the fungal biocontrol agent Beauveria bassiana alone or supplemented with an increasing sublethal rate of imidacloprid were conducted to quantify the fungal and chemical interactions on chrysanthemum aphid Macrosiphoniella sanborni (0.01‐0.05 a.i. μg mL^?1) and green peach aphid Myzus persicae (0.05‐0.5 a.i. μg mL^?1). During one week after exposure to a 1 mL spray onto a 95 cm² area in a Potter Spray Tower, M. sanborni was either more susceptible to B. bassiana or more sensitive to imidacloprid than M. persicae. The time–concentration–mortality (TCM) responses of each aphid species in each of five bioassays fit well to a TCM model, indicating a strong dependence of the fungal and chemical interactions on both concentration and post‐spray time. Adding imidacloprid to B. bassiana sprays at the rates of 0.025–0.05 μg mL^?1 against M. sanborni or 0.1–0.5 μg mL^?1 against M. persicae significantly enhanced or accelerated the fungal action. Based on the LC₅₀ or LC₉₀ estimates and their variances determined by the fitted TCM relationships for each aphid species, the relative potencies of an imidacloprid‐inclusive bioassay over those with B. bassiana alone or together with a lower sublethal rate ranged from a few to hundreds of times and varied over days after spray. These results suggest an alternative tactic for practical control of the aphid pests by a combined formulation or application of B. bassiana and imidacloprid and manage aphid resistance to the chemical insecticide.     

Cumulative impact of a clover cover crop on the persistence and efficacy of Beauveria bassiana in suppressing the pecan weevil (Coleoptera: Curculionidae)

The pecan weevil, Curculio caryae (Horn), is a key pest of pecan. Endemic levels of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin can occur in pecan orchards and contribute to natural control of C. caryae. Commercial formulations of the fungus can also be applied for suppression of C. caryae. We hypothesized that a clover cover crop enhances B. bassiana efficacy and persistence (e.g., by protecting the fungus against abiotic environmental stresses). The hypothesis was tested by conducting field trials in a pecan orchard in Byron, GA, in 2009 and 2010. The study included four treatments arranged in a factorial with two levels of fungus (endemic fungus only, and application of a commercial B. bassiana product), and two levels of clover (white clover, Trifolium repens L., and no clover). Fungal persistence was measured by determining the number of CFUs per gram of soil over time (during 42 d postapplication of B. bassiana in 2009 and 29 d in 2010). Efficacy was measured by capturing naturally emerging C. caryae and subsequently determining mortality and mycosis (over 24 d in 2009 and 17 d in 2010). In 2009, greater prevalence of B. bassiana conidia was detected in plots receiving fungal applications compared with no fungus applications, and no clear effect of clover was observed in plots receiving B. bassiana applications in either year. In 2010, B. bassiana prevalence in the endemic fungus plus clover treatment was higher than fungus without clover, and was similar to plots receiving additional B. bassiana applications. Given that we observed enhanced persistence of endemic B. bassiana in 2010 but not 2009, the impact of clover appears to be a cumulative effect. Mortality of C. caryae (averaged over the sampling periods) ranged between 68-74% in plots receiving B. bassiana applications and 51-56% in plots with endemic fungus only. C. caryae mortality and mycosis data also provided evidence that endemic B. bassiana efficacy was enhanced by clover relative to plots without clover (with no clear clover effect on plots receiving fungus applications). Thus, we conclude that natural control of C. caryae can increase when clover is grown in pecan orchards with endemic populations of B. bassiana.     

Regional and temporal variation in susceptibility to lambda-cyhalothrin in onion thrips, Thrips tabaci (Thysanoptera: Thripidae), in onion fields in New York

Populations of onion thrips, Thrips tabaci Lindeman, from commercial onion fields in New York were evaluated for their susceptibility to the commonly used pyrethroid, lambda-cyhalothrin (Warrior T), using a novel system called the Thrips Insecticide Bioassay System (TIBS). To use TIBS, thrips are collected directly from the plant into an insecticide-treated 0.5-ml microcentrifuge tube that has a flexible plastic cap with a small well into which 0.08 ml of a 10% sugar-water solution with food colorant is deposited. The solution is sealed into the well with a small piece of stretched parafilm through which the thrips can feed on the solution. Thrips mortality is assessed after 24 h with the help of a dissecting stereoscope. In 2001, onion thrips populations were collected from 16 different sites and resistance ratios were >1,000 in five populations. Percent mortality at 100 ppm, a recommended field rate, varied from 9 to 100%, indicating high levels of variation in susceptibility. Particular instances of resistance appeared to be the result of practices within an individual field rather than a regional phenomenon. In 2002, we also observed large differences in onion thrips susceptibility, not only between individual fields but also between thrips collected in a single field at mid season and late season, again suggesting that insecticide-use practices within an individual field caused differences in susceptibility. Additional tests indicated no differences in susceptibility between adult and larval onion thrips populations and only relatively minor differences between populations collected from different parts of the same field. Using TIBS, several populations of onion thrips with different susceptibilities to lambda-cyhalothrin were identified and then subjected to lambda-cyhalothrin-treated onion plants. There was a highly significant positive relationship between percent mortality of thrips from TIBS and percent mortality from the treated onion plants, indicating that results from TIBS could be used to predict spray performance. These data suggest that use of TIBS for evaluating susceptibility to particular insecticides could be instrumental for developing a resistance management strategy for onion thrips.     

Biological and chemical control of the Asiatic garden beetle, Maladera castanea (Coleoptera: Scarabaeidae)

The efficacy of chemical and biological control agents against larvae of the Asiatic garden beetle, Maladera castanea (Arrow), in turfgrass under laboratory, greenhouse, and field conditions were determined. In field trials where insecticides were applied preventively against eggs and young larvae, the molt-accelerating compound halofenozide and the neonicotinoids imidacloprid and thiamethoxam were ineffective, whereas another neonicotinoid, clothianidin, provided 62-93% control. In greenhouse experiments against third instars in pots, the carbamate insecticide carbaryl was ineffective, whereas the organophosphate trichlorfon provided 71-83% control. In laboratory, greenhouse, and field experiments, the entomopathogenic nematode Heterorhabditis bacteriophora Poinar and Steinernema glaseri Steiner (not tested in the field) were ineffective against third instars, whereas S. scarabaei Stock & Koppenh?fer provided excellent control. In microplot field experiments at a rate of 2.5 x 10(9) infective juveniles per ha, H. bacteriophora provided 12-33% control and S. scarabaei 71-86% control. Combinations of S. scarabaei and imidacloprid did not provide more control of third instars compared with S. scarabaei alone.     

Electroantennogram responses of male Sphinx perelegans hawkmoths to floral and ‘green-leaf volatiles’

The effects of sublethal dosages of the chloronicotinyl insecticide imidacloprid on different strains of the tobacco whitefly, Bemisia tabaci Gennadius (Homoptera: Aleyrodidae), have been studied after leaf dip and systemic application. All bioassays were performed with the insecticide susceptible strain, SUD-S, and two Spanish biotypes, ALM-2 and LMPA-2, both resistant to conventional insecticides and with a lower susceptibility towards imidacloprid. Honeydew, excreted by all strains feeding on treated and untreated cotton leaf discs was quantified by photometric analysis of its carbohydrate content. EC50-values for the depression of honeydew excretion in female adults after systemic application of imidacloprid were calculated at 0.037 ppm, 0.027 ppm and 0.048 ppm for strains SUD-S, ALM-2 and LMPA-2, respectively, indicating no significant differences between strains in feeding behaviour throughout an 48 h testing period. Depending on the strain these EC50-values were 150- to 850-times lower than LC50-values calculated for mortality in the same bioassay. Starvation tests revealed mean survival times of >48 h for female adults placed on agar without leaf discs, indicating that sublethal dosages of imidacloprid which caused antifeedant responses, were probably not covered in common 48 h systemic bioassays, used to monitor resistance to imidacloprid. Effects of sublethal dosages on honeydew excretion after leaf dip application seem to be minor. In choice situations with systemically treated and untreated leaf discs in a single container, female adults of B. tabaci showed a clear preference for the untreated leaf discs. However, when using leaf discs treated by painting the surface with imidacloprid in the same bioassay, feeding activities on treated and untreated leaf discs were not significantly different. The results of the present study demonstrate the antifeedant properties of imidacloprid on B. tabaci, which might play an essential role after soil application or seed treatment under field conditions.     

Entomopathogenic Fungi Associated with Natural Populations of the Moroccan Locust Dociostaurus maroccanus (Thunberg) (Orthoptera: Gomphocerinae) and Other Acridoidea in Spain

The entomopathogenic fungi Beauveria bassiana Vuill . and Metarhizium anisopliae (Metschnikoff) have been found in natural populations of the Moroccan locust Dociostaurus maroccanus (Thunberg) and other species of acridoids that cohabit the same locust breeding areas in southern Spain . Infection levels of B. bassiana on insects collected in the field and maintained under laboratory conditions were relatively high (1 . 6 - 20 . 5%) . The prevalence of the disease extended from spring to summer in the three consecutive years monitored . A local isolate of this fungus demonstrated high virulence (LD = 440 conidia / insect) against the 50 locust D. maroccanus in the laboratory bioassay . The relatively wide host range of B. bassiana and its adaptation to the dry and hot conditions dominating the locust breeding area suggest that this isolate could be considered in the development of a biological control programme against D. maroccanus.     

Beauveria bassiana (Balsamo) Vuillemin - a Promising Microbial Control Agent of the Satin Moth (Lepidoptera: Lymantriidae)

A new isolate of the entomopathogenic fungus, Beauveria bassiana , was tested by laboratory bioassays for potential use as a microbial control agent of the early (third) and last (seventh) instars of the satin moth, Leucoma salicis L. Results demonstrate that this fungus is very effective against last instars of this lepidopteran pest. Although B. bassiana was cultured successfully from internal contents of all cadavers examined, vegetative hyphae emerged from intact cadavers of L. salicis infected as early (third) instar caterpillars only.     

Combined effect of microbial and chemical control agents on subterranean termites

Termite mortality was measured when fungi were combined with bacteria or a chemical termiticide to determine whether a synergistic effect occurred. The fungus Beauveria bassiana was combined with the non-repellant chemical termiticide imidacloprid. Of the three B. bassiana strains tested one, B. bassiana ATCC 90519, was sufficiently pathogenic on its own that the advantage of a supplementary chemical treatment was marginal. The mortality caused by another fungal strain, B. bassiana ATCC 26037, was improved in combination with imidacloprid at both of the tested chemical concentrations over the first 14 days. The remaining fungal strain, B. bassiana ATCC 90518, demonstrated an overall mortality rate in combination with imidacloprid of 82.5%, versus a rate of 65.0% for the fungus alone. The fungus Isaria fumosorosea (Ifr) was combined with the bacterium Bacillus thuringiensis (Bt). On day 5, Ifr, Bt, and the combined treatment at a 10⁶ spores or cells/ml dosage caused 8.8%, 22.5%, and 15.0% mortality, respectively. The Bt and combined mortality rates are not significantly different. Control mortality on day 5 was 5.0%. On day 13 the combined 10⁶ treatment mortality rate was 91.3%, which was significantly higher than all other treatments: control at 17.5%, Ifr at 36.3% and Bt at 35.0%. When Ifr and Bt were applied at a 10⁹ spores or cells/ml dosage, Ifr alone caused a mortality rate of 97.5% as early as day 5. The combination with Bt could not significantly increase the effectiveness of this dosage. These data demonstrate the potential for synergistic effects of fungal and chemical treatment methods, thereby broadening the use of microbial control agents and reducing the quantity of chemical agents necessary to effect control.