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.     

Interaction effects between Beauveria bassiana and imidacloprid against Thrips tabaci (Thysanoptera: Thripidae)

The onion thrips, Thrips tabaci Lindeman is a major pest of many horticultural crops in many parts of the world. The entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin and imidacloprid are used for the control of the onion thrips. The potential interactions between the fungus and the chemical against the onion thrips adults were evaluated in a laboratory and two greenhouse bioassays. The laboratory bioassay was a leaf dip bioassay for a mixture of B. bassiana and imidacloprid. The first greenhouse bioassay was a topical application of a mixture of the fungus and the chemical while the second bioassay was a drenching application of imidacloprid and a topical application of B. bassiana. In all bioassays, the combined application resulted in higher percentage mortalities than either B. bassiana or imidacloprid alone. In the laboratory bioassay, the highest mortality of 97% was achieved by using B. bassiana at the field rate with imidacloprid at 1/10th the field rate relative to 88%, 94%, 21% for B. bassiana alone, imidacloprid alone and the control, respectively. In the first greenhouse bioassay, the highest mortality of 80% resulted from the combined application of B. bassiana and imidacloprid at field rates compared with 55%, 75%, and 22% for B. bassiana alone, imidacloprid alone and the control, respectively. In the second greenhouse bioassay, the combined application of both control agents at field rates resulted in 85% mortality compared with 52%, 83%, and 18% for B. bassiana alone, imidacloprid alone and the control, respectively. The interaction effect between the two control agents was additive except when B. bassiana at 1/2 field rate was mixed with imidacloprid at 1/10th field rate in the laboratory bioassay, where the interaction was antagonistic. The above results indicate that combining imidacloprid with B. bassiana might reduce the rate of the insecticide application while enhancing the efficacy of the biological control agent.     

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.     

Effect of combining imidacloprid and diatomaceous earth with Beauveria bassiana on mole cricket (Orthoptera: Gryllotalpidae) mortality

Sublethal doses of three orthopteran-derived strains of Beauveria bassiana (Balsamo) Vuillemin were topically applied to adult southern mole crickets, Scapteriscus borellii Giglio-Tos (Orthoptera: Gryllotalpidae), and tested in combination with substrate treatments of diatomaceous earth (DE) and imidacloprid. Crickets treated only with the high doses (10(8) conidia per cricket) of each of the three B. bassiana strains exhibited the shortest survival times as well as the highest percentage mortality at 28 d after treatment. However, these treatments did.not differ significantly from any of the diatomaceous earth combination treatments. Two of the strains tested, 5977 and 3622, exhibited synergistic interactions with DE, whereas the third strain, GHA, was not significant for synergy. Mortality caused by the combination treatment was still greater than the expected additive effect. DE abrades the insect cuticle and absorbs cuticular lipids, aiding the entry of germinating conidia into the mole cricket hemocoel. None of the three strains exhibited synergy when combined with imidacloprid, and mortality of all combination treatments was less than additive. For strain 5977, there was an antagonistic interaction with imidacloprid. It was difficult to obtain <30% mortality for imidacloprid only treatments, which was considered the upper limit for sublethal doses. The mean percentage mortality caused by imidacloprid was 37.5%, and this high percentage made it difficult for any combination treatment to cause significantly more mortality than the expected additive effect. These results clarify the interactions of other control products with B. bassiana and provide a basis for a reduced pesticide approach to mole cricket control.     

两种杀虫真菌制剂与低用量吡虫啉对温室粉虱的协同防效评价

在昆明市郊蔬菜基地4间大棚(5m×100m·间^-1)栽培的生菜上,用球孢白僵菌(Beauveriabassiana)和玫烟色拟青霉(Paecilomyces fumasonoseus)的纯孢子乳悬剂(有效含孢量10^10·ml^-1)及其与1％和3％(w／v)的吡虫啉10％WP的混配剂对温室粉虱(Trialeurodes vaporariorum)进行了协同药效试验,吡虫啉添加量仅相当于推荐用量的6．4％～9．5％和19．1％～28．6％,试验含6个菌剂处理、2个低用量吡虫啉处理和1个清水对照,各有3个小区(7m×5m·小区^-1)重复,随机区组排列,稀释1000倍的菌液(含孢量10^7·ml^-1)和吡虫啉液于生菜移栽后第11d首次喷雾,15d后喷第二次,首次用菌前调查初始粉虱密度,用菌后每隔5d调查活虫数和死虫数,从8月20日起历时30d,结果表明,连续2次用菌有效地控制了粉虱对当季大棚生菜的危害,各菌剂处理的最终相对防效和虫口减退率均达到95％以上,极显著地高于低用量吡虫啉处理,其中,玫烟色拟青霉制剂对粉虱的控制效果一般优于球孢白僵菌制剂,且与吡虫啉的添加量呈正相关,但是,各菌剂处理间控虫效果的差异主要发生在首次用菌后第5d和10d,此后差异逐渐缩小,至第二次用菌后第10d各菌剂处理间无显著差异,本试验结果表明,两种真菌的孢子乳悬剂均可用于温室粉虱的防治,添加微量吡虫啉可增强菌剂的防效。     

Cuticular lipids and silverleaf whitefly stage affect conidial germination of Beauveria bassiana and Paecilomyces fumosoroseus

Beauveria bassiana and Paecilomyces fumosoroseus are generalist entomopathogenic fungi that infect the silverleaf whitefly (Bemisia argentifolii). We found second and third instar whiteflies to be the most susceptible larval stage to both fungi. Conidia of B. bassiana germinated most readily on the cuticle of second instars (54% germinated) and P. fumosoroseus germination was highest on third instar cuticle (45%). Fourth instars (the ultimate instar) had low susceptibility to these pathogens, and spore germination on the cuticle of fourth instars was very low for B. bassiana (7%) and intermediate for P. fumosoroseus (33%). Cuticular lipids were found to have toxic or inhibitory effects on conidia of B. bassiana and P. fumosoroseus when the spores were germinated on nutrient agar in the presence of the lipids. In the absence of added nutrients, P. fumosoroseus conidial germination increased in the presence of the lipids. To test if the inhibitory effects of the lipids were due solely to hydrophobicity (preventing water from coming into contact with the conidia) we tested the effects of synthetic long-chain wax esters. The synthetic wax esters inhibited germination of P. fumosoroseus to a degree that was similar to the effect of the cuticular lipid extracts, but the synthetic lipids did not have a significant effect on B. bassiana. Thus, the thick coating of long-chain wax esters produced by whitefly nymphs affect spore germination of fungal pathogens, but whether they play a significant role in defense against disease is not clear.     

Recombinant immunosuppressive protein from Pimpla hypochondrica venom (rVPr1) increases the susceptibility of Mamestra brassicae larvae to the fungal biological control agent, Beauveria bassiana

Although fungi are used to control a variety of insect pests, it is accepted that their usage could be increased if their efficacy was greater. The outcome of the interaction of a fungus and a pest insect may be influenced by a number of criteria, including the ability of the insect to mount effective immune responses against the pathogen. In view of this, we aimed to determine if a recombinant immunosuppressive wasp venom protein (rVPr1) can increase the susceptibility of larvae of the lepidopteran pest, Mamestra brassicae, to the fungal biological control agent, Beauveria bassiana. Bioassays indicated that when larvae were injected with 3.5 μl of rVPr1 and 100 B. bassiana conidia (combined injection assays), a significant reduction in survival of larvae occurred compared with each treatment on its own (P=0.006). Similar results were obtained when larvae were dipped in a solution containing 3 × 10(6) B. bassiana conidia per ml and then injected with 3.5 μl of rVPr1 2 days later (topical application assays), (P<0.001). These results indicate that rVPr1 can increase the efficacy of B. bassiana toward a lepidopteran pest, and are discussed within the context of insect immune responses and integrated pest management.     

马尾松林中球孢白僵菌寄主转移和专化性的SSR标记分析

球孢白僵菌是一种最常见的虫生真菌,已被广泛应用于害虫生物防治。利用SSR简单序列重复(simple sequence repeat,SSR)分子标记对来自安徽省麻姑山马尾松林的102株球孢白僵菌进行基因分型、寄主转移和寄主专化性研究。9对微卫星引物将102株白僵菌分成31个微卫星基因型。在这31个微卫星基因型中,有5种基因型株系为相对优势菌株,这5种基因型株系通过侵染不同寄主昆虫即寄主转移延续自身在马尾松林生态系统中的传播和流行,从而证实寄主转移是白僵菌群体中的普遍现象。同时,这些相对优势基因型并不是在各个月份均匀分布,在大部分月份中,存在1–2种优势度高的基因型。相同基因型株系可侵染不同寄主的结果揭示出白僵菌不仅在种的水平,而且在菌株水平上寄主专化性也较弱。正是白僵菌较弱的寄主专化性特征促使部分基因型株系通过寄主转移,在马尾松林生态系统中得以宿存和延续。     

Effects of exogenous nutrients on conidial germination and virulence against the silverleaf whitefly for two hyphomycetes

Exogenous protein and sugar sources were tested for their impact on conidial germination of two silverleaf whitefly (Bemisia argentifolii) pathogens: Beauveria bassiana and Paecilomyces fumosoroseus. In liquid culture, sugars stimulated only 5-27% germination of B. bassiana and < or =11% germination of P. fumosoroseus, whereas, yeast extract or peptone stimulated 95-100% germination. In the absence of additional nutrients, agar alone stimulated approximately 50% germination. Storing spores for different periods of time did not alter their general response to exogenous nutrients. When spores were germinated before being applied to third instar B. argentifolii, mortality was as much as 2.45 times greater and occurred more rapidly than that for fresh spores. For ungerminated conidia, the mean time to death from infection was 5.45 (SE = 0.16) and 4.74 (SE = 0.08) days for application rates of 37 and 144 conidia x mm(-2), respectively. When conidia were germinated before application, infection times dropped to 4.58 (SE = 0.16) and 4.45 (SE = 0.10) days, respectively. A likely explanation for the greater pathogenicity and virulence of germinated over ungerminated B. bassiana conidia is that only a fraction of the spores applied to whitefly nymphs actually germinate on the cuticle. For some specialized applications, such as greenhouse production systems, it may be beneficial to germinate spores immediately prior to application.     

Increased insect virulence in Beauveria bassiana strains overexpressing an engineered chitinase

Entomopathogenic fungi are currently being used for the control of several insect pests as alternatives or supplements to chemical insecticides. Improvements in virulence and speed of kill can be achieved by understanding the mechanisms of fungal pathogenesis and genetically modifying targeted genes, thus improving the commercial efficacy of these biocontrol agents. Entomopathogenic fungi, such as Beauveria bassiana, penetrate the insect cuticle utilizing a plethora of hydrolytic enzymes, including chitinases, which are important virulence factors. Two chitinases (Bbchit1 and Bbchit2) have previously been characterized in B. bassiana, neither of which possesses chitin-binding domains. Here we report the construction and characterization of several B. bassiana hybrid chitinases where the chitinase Bbchit1 was fused to chitin-binding domains derived from plant, bacterial, or insect sources. A hybrid chitinase containing the chitin-binding domain (BmChBD) from the silkworm Bombyx mori chitinase fused to Bbchit1 showed the greatest ability to bind to chitin compared to other hybrid chitinases. This hybrid chitinase gene (Bbchit1-BmChBD) was then placed under the control of a fungal constitutive promoter (gpd-Bbchit1-BmChBD) and transformed into B. bassiana. Insect bioassays showed a 23% reduction in time to death in the transformant compared to the wild-type fungus. This transformant also showed greater virulence than another construct (gpd-Bbchit1) with the same constitutive promoter but lacking the chitin-binding domain. We utilized a strategy where genetic components of the host insect can be incorporated into the fungal pathogen in order to increase host cuticle penetration ability.     

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.     

Community composition, host range and genetic structure of the fungal entomopathogen Beauveria in adjoining agricultural and seminatural habitats

Although intensively investigated for biological control of insect pests, little is known about the ecology of the fungal entomopathogenic genus Beauveria in natural or agricultural habitats. In this study, we used molecular phylogenetic and genotypic information to infer species diversity, reproductive potential and genetic structure of Beauveria occurring within a single arable field and bordering hedgerow in Denmark. Isolates were sampled from cultivated field and hedgerow soils, from insects harbouring latent fungal infections, and from the phylloplanes of three plant species common in the hedgerow flora. A nuclear phylogeny of this local Beauveria assemblage resolved seven phylogenetic species, including (i) five phylogenetic species within Beauveria bassiana sensu stricto ; (ii) Clade C, a taxonomically uncharacterized species that is morphologically indistinguishable but phylogenetically distant from B. bassiana s.s. ; and (iii) Beauveria brongniartii. All seven species were present throughout the hedgerow habitat, including as infections in insects. Significantly, only B . bassiana s.s. phylogenetic species Eu_1 was isolated from tilled soils. Mating type polymerase chain reaction assays demonstrated that all five B. bassiana s.s. phylogenetic species possess bipolar outcrossing mating systems. Of these, only the Eu_1 population contained two mating types; however, a 31:2 skew in MAT1:MAT2 mating types suggests a low frequency of sexual reproduction in this population. The four remaining B. bassiana s.s. phylogenetic species were fixed for single mating types and these populations are evidently clonal. Multilocus microsatellite genotyping revealed polymorphism in all five phylogenetic species of B. bassiana s.s. ; however, all show evidence of clonal genetic structure.     

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).     

Molecular investigation on strain genetic relatedness and population structure of Beauveria bassiana

Triplicate molecular methods, i.e. polymerase chain reaction-restriction fragment length polymorphism of the pr1 gene, microsatellite markers and 28S rDNA haplotyping by detecting the presence or absence of group I introns, were used for population study of the entomopathogenic fungus, Beauveria bassiana. The findings showed that the average genetic diversity index of geographical populations was significantly smaller than that of populations derived from insect host orders, indicating that the genetic relatedness of B. bassiana strains was highly associated with geographical locality rather than insect host species. The reproductive style of all the B. bassiana populations was found to be non-clonal. Population structure analysis revealed that the average divergent coefficient among populations of B. bassiana was far below 1 (0.1112), which indicated that there was no significant genetic differentiation between populations, and that the overall genetic diversity mainly resulted from the genetic variations within geographical populations. Statistically, genetic distances between populations were positively correlated with geographical distances, suggesting that geographical separation poses an obstacle to the possibility and frequency of genetic exchanges between populations. On the other hand, gene flow was indirectly established to occur between B. bassiana populations.     

A Beauveria phylogeny inferred from nuclear ITS and EF1-alpha sequences: evidence for cryptic diversification and links to Cordyceps teleomorphs

Beauveria is a globally distributed genus of soil-borne entomopathogenic hyphomycetes of interest as a model system for the study of entomopathogenesis and the biological control of pest insects. Species recognition in Beauveria is difficult due to a lack of taxonomically informative morphology. This has impeded assessment of species diversity in this genus and investigation of their natural history. A gene-genealogical approach was used to investigate molecular phylogenetic diversity of Beauveria and several presumptively related Cordyceps species. Analyses were based on nuclear ribosomal internal transcribed spacer (ITS) and elongation factor 1-alpha (EF1-alpha) sequences for 86 exemplar isolates from diverse geographic origins, habitats and insect hosts. Phylogenetic trees were inferred using maximum parsimony and Bayesian likelihood methods. Six well supported clades within Beauveria, provisionally designated A-F, were resolved in the EF1-alpha and combined gene phylogenies. Beauveria bassiana, a ubiquitous species that is characterized morphologically by globose to subglobose conidia, was determined to be non-monophyletic and consists of two unrelated lineages, clades A and C. Clade A is globally distributed and includes the Asian teleomorph Cordyceps staphylinidaecola and its probable synonym C. bassiana. All isolates contained in Clade C are anamorphic and originate from Europe and North America. Clade B includes isolates of B. brongniartii, a Eurasian species complex characterized by ellipsoidal conidia. Clade D includes B. caledonica and B. vermiconia, which produce cylindrical and comma-shaped conidia, respectively. Clade E, from Asia, includes Beauveria anamorphs and a Cordyceps teleomorph that both produce ellipsoidal conidia. Clade F, the basal branch in the Beauveria phylogeny includes the South American species B. amorpha, which produces cylindrical conidia. Lineage diversity detected within clades A, B and C suggests that prevailing morphological species concepts underestimate species diversity within these groups. Continental endemism of lineages in B. bassiana s.l. (clades A and C) indicates that isolation by distance has been an important factor in the evolutionary diversification of these clades. Permutation tests indicate that host association is essentially random in both B. bassiana s.l. clades A and C, supporting past assumptions that this species is not host specific. In contrast, isolates in clades B and D occurred primarily on coleopteran hosts, although sampling in these clades was insufficient to assess host affliation at lower taxonomic ranks. The phylogenetic placement of Cordyceps staphylinidaecola/bassiana, and C. scarabaeicola within Beauveria corroborates prior reports of these anamorph-teleomorph connections. These results establish a phylogenetic framework for further taxonomic, phylogenetic and comparative biological investigations of Beauveria and their corresponding Cordyceps teleomorphs.     

Diatomaceous earth increases the efficacy of Beauveria bassiana against Tribolium castaneum larvae and increases conidia attachment

This research tested the suppressive ability of Beauveria bassiana (Balsamo) Vuillemin alone and in combination with diatomaceous earth against the red flour beetle, Tribolium castaneum (Herbst). Adults did not show a dose response to B. bassiana, and the addition of diatomaceous earth (DE) did not result in a significant increase in mortality. Against larvae, however, DE at 190 mg/kg grain enhanced the efficacy of B. bassiana at all concentrations ranging from 33 to 2,700 mg of conidia per kilogram of grain. The presence of DE resulted in 17- and 16-fold decreases in the median lethal concentration of B. bassiana at 56 and 75% RH, respectively. No significant differences in larval mortality in response to B. bassiana and diatomaceous earth alone or in combination were found between 56 and 75% RH. Conidial attachment to larvae was significantly greater with 190 mg/kg DE than without it. The partial analysis of lipids taken up by DE from the larvae revealed the removal of phospholipids and long-chain fatty acids. These results support the hypothesis that diatomaceous earth enhances the efficacy of B. bassiana against larval T. castaneum, at least in part by damaging the insect cuticle, thus increasing conidial attachment and making nutrients more available to conidia for their germination.     

Synergistic effect of entomogenous fungi on some insecticides against Bihar hairy caterpillar Spilarctia obliqua (Lepidoptera: Arctiidae)

A number of fungal parasites infect a wide range of insects and cause epizootics from time to time. Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metschnikoff) Sorokin are two of the major disease-causing fungi in insects. Investigations were carried out to study the effect of these fungi on the toxicity of endosulfan, imidacloprid, lufenuron, diflubenzuron, dimethoate and oxydemeton methyl against 10-11 days old larvae of Spilarctia obliqua (Walker). For some products the combination treatments showed higher dose mortality response than the sole treatment of fungal conidia or the insecticide. The combination of insecticides with B. bassiana showed 1.26-35.8 fold increase in toxicity of insecticides over sole treatment, while the increase was 1.05-72.0 fold in case of M. anisopliae. Imidacloprid 17.8 SL and oxydemeton methyl 25EC may be used in combination with these fungi for management of S. obliqua.     

Compatibility of the insect pathogenic fungus Beauveria bassiana with neem against sweetpotato whitefly, Bemisia tabaci, on eggplant

A study on the compatibility of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) with neem was conducted against sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), on eggplant. Initially, three concentrations of B. bassiana (10⁶, 10⁷, and 10⁸ conidia ml⁻¹) and three concentrations of neem (0.25, 0.5, and 1.0%) were used as individual treatments against B. tabaci . The highest concentration of B. bassiana yielded the highest B. tabaci egg (25.2%) and nymph mortalities (73.0%), but this was not significantly different from the mortalities caused by the 10⁷ conidia ml⁻¹ suspension. Similarly, the highest concentration of neem yielded the highest egg (27.3%) and nymph mortalities (75.5%), which was also not significantly different from the 0.5% suspension. Therefore, 0.5% neem was used along with 10⁷ B. bassiana conidia ml⁻¹ suspension as an integrated pest management program against B. tabaci . The combination of B. bassiana and neem yielded the highest B. tabaci egg (29.5%) and nymph mortalities (97.2%), and the lowest LT₅₀ (2.08 day) value. Moreover, an integrated combination of B. bassiana with neem caused 27.6 and 20.5% more nymphal mortality than individual treatments of B. bassiana and neem, respectively, 7 days post-application. Thus, a combined application of an entomopathogenic fungus and a botanical insecticide may benefit from both, and it has proven effective for the control of B. tabaci on eggplant.     

Populational parameters of Spalangia endius Walker (Hymenoptera: Pteromalidae) on Pupae of Musca domestica L. (Diptera: Muscidae) treated with two strains of Beauveria bassiana (Bals.) Vuil. (Deuteromycetes)

The parasitoid Spalangia endius Walker is an efficient controller of Dipteran pupae, such as Musca domestica L. The entomopathogenic fungus Beauveria bassiana (Bals.) Vuil. is a regulator of insect populations, including these synanthropic pests. The aim of this work was to explore the possibilities of utilizing both agents in a combined form for the biocontrol of the domestic fly. Recently formed M. domestica pupae were inoculated by immersion in conidia suspension (10(8) conidia/ml) with two strains of B. bassiana (Bb6 and Bb10). The inoculated pupae were offered to the female parasitoid. In one bioassay they were offered pupae inoculated a single day and in other, pupae inoculated the following day as well. In both bioassays non inoculated (control) pupae were offered to the parasitoids until their death. Thirty females of S. endius were used for each strain and bioassay. From the study of the parasitoid offspring, life tables were built and the reproduction net rate (R(0)) and intrinsic natural increase (r(m)) were obtained among other demographic parameters; the parasitism percentages and sex ratios were also analyzed. B. bassiana did not affect significantly the biodemography of the parasitoid when pupae were inoculated a single time. On the other hand the R0 and the rm were smaller than that of the control without the fungus when pupae were inoculated twice, although sporulation was not observed in the cadavers of S. endius.