Infected Aphids Research Articles

Prevalence of Pandora neoaphidis (Zygomycetes: Entomophthorales) infecting Nasonovia ribisnigri (Hemiptera: Aphididae) on lettuce crops in Argentina

Lettuce crops, Lactuca sativa, organically produced in La Plata, Argentina, were sampled in order to determine the prevalence of fungal diseased aphids. Nasonovia ribisnigri was the only aphid detected and its occurrence was highly variable. The fungal pathogen Pandora neoaphidis (Entomophthoromycotina: Entomophthorales) was the only pathogen detected. We recorded a maximum of 34.2 aphids per plant and the highest rate of fungal prevalence was 56.6% ( n = 30) (aphids infected/total aphids). Infected aphids were observed in all sampling sites. No differences of infection rates were detected between the center and the edge of crops. Host density was an important factor determining infection. The majority of host population was comprised of nymphs which were the most infected in terms of individuals per habitat unit (lettuce plant), but considering the proportion of infected aphids per stage of development, the prevalence of infection in nymphs and adults was similar.

Characterization and virulence of Lecanicillium lecanii against different aphid species

Seven isolates of Lecanicillium lecanii (Zimmermann) Zare & Gams isolated in Spain from infected aphids were characterized using sequences of the Internal Transcribed Spacer (ITS) regions and also based on morphological and physiological characteristics. Four of these seven L. lecanii isolates were selected to assess their virulence against nymphs of Myzus persicae (Sulzer), Nasonovia ribisnigri (Mosley), Macrosiphum euphorbiae (Thomas) and Aphis gossypii Glover. Mortality (%), lethal concentration 50 (LC50) and lethal time 50 (TC50) were calculated. The analysis of the sequences of ITS region confirmed that the new isolates were clearly Lecanicillium lecanii. The set of isolates had similar radial growth (51.5–54.0 mm), except for ICAL1 (39 mm). The germination time 50 (GT50) varied between 10.7 h (ICAL3) and 13 h (ICAL5). The isolate ICAL6 showed the highest value for conidial production (3.4 × 108 con ml−1) and also produced the highest mortality for M. persicae (95%) and was more virulent than the commercial product Vertalec® (91.6%).

Laboratory and field evaluations of entomopathogenic Lecanicillium attenuatum CNU-23 for control of green peach aphid (Myzus persicae).

An entomopathogenic fungus was isolated from an infected aphid. The isolate conformed most closely to Lecanicillium attenuatum CBS 402.78 (AJ292434) based on the internal transcribed spacer (ITS) region of its 18S rDNA, and thus was designated L. attenuatum CNU-23. Laboratory and field evaluations of CNU-23 blastospores were carried out for the control of green peach aphids. The laboratory evaluations of CNU-23 revealed an aphid mortality of about 80% with an estimated LT50 of 3.72 days after the application of CNU-23 at 1x106 blastospores/ ml. Meanwhile, the field evaluations of CNU-23 performed on greenhouse pepper plants during the rainy season showed an aphid mortality ranging from 72% to 97%. Significant sporulation was observed in the aphids treated with CNU-23. Therefore, the results suggest that L. attenuatum CNU-23 can be used as a biocontrol agent for green peach aphids on greenhouse pepper plants.

Aislamiento de Conidiobolus coronatus a partir de una plaga de áfidos del cebollín ( Allium schoenoprasum L.)

Conidiobolus coronatus (Entomophthorales), a pathogen of human and some insects, was isolated from dead specimens of the aphid Neotoxoptera formosana present on chives leaves. C. coronatus infected aphids could pose a threat to growers health. Pest control strategies of N. formosana may also contribute to reduce the human infection risk by C. coronatus.

Grain aphid population structure: no effect of fungal infections in a 2‐year field study in Denmark

Abstract1 Sitobion avenae (F.) is a serious pest in Danish cereal crops. To understand the population genetic structure, aphids were sampled in seven different winter wheat (Triticum sativum Lamarck) fields throughout Denmark. The aphids were genotyped with seven microsatellite markers. In total, 2075 aphids were collected and 1203 of these were genotyped.2 The Danish S. avenae populations displayed very high genotypic diversity, high percentages of unique genotypes and low linkage disequilibria; this is likely to be a result of genetic recombination encompassed by their holocyclic lifestyle. The populations showed very limited differentiation and no sign of isolation by distance. Almost all the genetic variation was ascribed within the populations rather than between populations, probably due to a high migration rate at approximate 10% per generation.3 Seasonal changes in clonal diversity and distribution of asexual summer generations of S. avenae within the infestation period in a single winter wheat field were followed over two consecutive years by weekly sampling from 60 plots each of 20 × 20 m. Clonal diversity was high in all samples with no dominant clonal lineages and no significant difference in the genotypic diversity between weeks or between years. However, a temporal genetic differentiation effect, throughout the infestation, suggests that selective factors or high temporal migration play an important role in shaping the genetic structure S. avenae.4 Analyses of fungal infected and uninfected aphids were performed to test whether some clonal linage were more often infected by fungi from the Entomophthorales under field conditions. In total, 54 progeny from aphids with Entomophthorales were genotyped and compared with 422 uninfected aphid genotypes. The Entomophthorales‐infected aphid genotypes did not cluster out together, suggesting that these fungal pathogens did not affect the population differentiation or clonal distribution of S. avenae in a Danish agroecosystem.5 Our findings indicate that S. avenae populations can be controlled using conservation biological control

A PCR-based tool for the cultivation-independent monitoring of Pandora neoaphidis

Pandora neoaphidis is one of the most important fungal pathogens of aphids and has a great potential for use in biocontrol. Little is known on how this fungus persists in an area and in particular on its overwintering strategies. It is hypothesized that natural areas play an important role for survival and that soil may serve as a source of inoculum for new aphid populations in spring. To test these hypotheses, a cultivation-independent PCR-based diagnostic tool was developed, that allows the detection of P. neoaphidis in the environment. Two P. neoaphidis specific PCR primer pairs were designed, targeting sequences in the ribosomal RNA gene cluster. Specificity of both primer pairs was demonstrated with P. neoaphidis and non-target close entomophthoralean relatives. Moreover, single amplicons of expected sizes were obtained with both primer pairs from various environmental sample types, including aphid cadavers, plant material, and soil. The PCR-based diagnostic tool was applied to investigate the persistence of P. neoaphidis in soil samples obtained in 2004/2005 from a nettle field harboring infected aphids in fall 2004. P. neoaphidis was detected in every sample collected in November 2004 and March 2005, suggesting an overwintering stage of P. neoaphidis in top soil layers. The developed cultivation-independent PCR-based tool will be valuable for further investigation of the ecology of P. neoaphidis and for the development and future implementation of management strategies against aphids involving conservation biocontrol.

Infection with an insect virus affects olfactory behaviour and interactions with host plant and natural enemies in an aphid

AbstractAphid ecology and population dynamics are affected by a series of factors including behavioural responses to ecologically relevant chemical cues, capacity for population growth, and interactions with host plants and natural enemies. Using the aphid Rhopalosiphum padi (L.) (Homoptera: Aphididae), we showed that these factors were affected by infection with Rhopalosiphum padi virus (RhPV). Uninfected aphids were attracted to odour of uninfected aphids on the host plant, an aggregation mechanism. However, infected aphids were not attracted, and neither infected nor uninfected aphids were attracted to infected aphids on the plant. Infected aphids did not respond to methyl salicylate, a cue denoting host suitability. Infected aphids were more behaviourally sensitive to aphid alarm pheromone, and left the host plant more readily in response to it. RhPV reduced the lifespan and population growth rate of the aphid. The predacious ladybird, Coccinella septempunctata (L.) (Coleoptera: Coccinellidae), consumed more infected aphids than uninfected aphids in a 24‐h period, and the aphid parasitoid Aphidius ervi Haliday (Hymenoptera: Aphidiidae) attacked more infected than uninfected aphids. However, the proportion of mummies formed was lower with infected aphids. The results represent further evidence that associated organisms can affect the behaviour and ecology of their aphid hosts.

Intraguild predation of the aphid pathogenic fungus Pandora neoaphidis by the invasive coccinellid Harmonia axyridis

Abstract 1. The ladybird Harmonia axyridis is an invasive alien species in many countries and is predicted to have a negative impact on native biodiversity. However, little is known on the status of this aphidophage as an intraguild predator of natural enemies of aphids such as insect‐pathogenic fungi.2. The study assessed the predation of the aphid‐specific pathogenic fungus Pandora neoaphidis by adult and larval H. axyridis collected from the U.K. (an invasive population) and Japan (a native population) relative to that of the ladybird Coccinella septempunctata (native to the U.K.) and the non‐U.K. C. septempunctata subspecies brucki that were either starved or unstarved.3. Overall, predation of uninfected aphids was greater than infected aphids and, when given a choice, a preference for aphids was shown. However, the extent of this preference was dependent on the species and origin of the coccinellid. Harmonia axyridis (U.K.) consumed the greatest quantity of fungal cadavers and showed little preference for uninfected aphids over infected aphids. In contrast, C. septempunctata rarely consumed infected aphids. Life stage had no direct effects on predation but starved coccinellids consumed more uninfected aphids than infected aphids.4. Harmonia axyridis (U.K.) is a stronger intraguild predator of P. neoaphidis cadavers than the native species C. septempunctata and, therefore, may have an impact on the occurrence and persistence of P. neoaphidis. The differences in intraguild predation by H. axyridis collected in the U.K. and those from Japan suggests that individuals that invaded the U.K. could have a different genetic profile to those in its native range.

Population dynamics of defensive symbionts in aphids

Vertically transmitted micro-organisms can increase in frequency in host populations by providing net benefits to hosts. While laboratory studies have identified diverse beneficial effects conferred by inherited symbionts of insects, they have not explicitly examined the population dynamics of mutualist symbiont infection within populations. In the pea aphid, Acyrthosiphon pisum, the inherited facultative symbiont, Hamiltonella defensa, provides protection against parasitism by the wasp, Aphidius ervi. Despite a high fidelity of vertical transmission and direct benefits of infection accruing to parasitized aphids, Hamiltonella remains only at intermediate frequencies in natural populations. Here, we conducted population cage experiments to monitor the dynamics of Hamiltonella and of another common A. pisum symbiont, Serratia symbiotica, in the presence and absence of parasitism. We also conducted fitness assays of Hamiltonella-infected aphids to search for costs to infection in the absence of parasitism. In the population cages, we found that the frequency of A. pisum infected with Hamiltonella increased dramatically after repeated exposure to parasitism by A. ervi, indicating that selection pressures from natural enemies can lead to the increase of particular inherited symbionts in insect populations. In our laboratory fitness assays, we did not detect a cost to infection with Hamiltonella, but in the population cages not exposed to parasitism, we found a significant decline in the frequency of both Hamiltonella and Serratia. The declining frequencies of Hamiltonella-infected aphids in population cages in the absence of parasitism indicate a probable cost to infection and may explain why Hamiltonella remains at intermediate frequencies in natural populations.

Predation rate and development of Coccinella septempunctata L. influenced by Neozygites fresenii-infected cotton aphid prey

Laboratory studies were conducted to determine the effect of cotton aphids [ Aphis gossypii Glover (Homoptera: Aphididae)] infected with Neozygites fresenii (Nowakowski) Batko (Entomophthorales: Neozygitaceae), on the number of prey attacked by and development of Coccinella septempunctata L. (Coleoptera: Coccinellidae). A diet of N. fresenii-infected aphids (in early stages of infection) did not have a significant effect on predation rate by either the fourth-stage larvae or adults of C. septempunctata. Second-, third- and fourth-stage larvae of C. septempunctata reared on fungus-infected aphids had significantly longer stadia than those reared on uninfected aphids. Mortality of C. septempunctata larvae reared on fungus-infected aphids increased between the second and the fourth instars, whereas mortality for those reared on uninfected aphids was significantly lower than those reared on infected aphids. Feeding on N. fresenii-infected aphids resulted in significantly smaller body size of C. septempunctata adults and a corresponding reduction in the number of eggs oviposited during a 29-day period relative to those fed a diet of uninfected aphids. Although our findings suggest that a diet of N. fresenii-infected aphids had no effect on the number of prey consumed by C. septempunctata, it had a significant effect on the development of the predator and its capacity to reproduce. These fitness costs could alter the capacity of the predator population to reduce subsequent pest populations in cotton or adjacent crops.

The effect of aphid infection and cultivar on the protein content governing baking quality of wheat flour

AbstractBACKGROUND: Aphids cause both quantitative and qualitative yield losses in cereals. Aphid feeding resulted in a significant decrease in gliadin/glutenin ratio in soft‐kernel wheat. Effect of aphid feeding on hard‐kernel wheat was examined.RESULTS: Plants of two different hard‐wheat cultivars were artificially infected by two aphid species: Sitobion avenae and Diuraphis noxia. Three different levels of caging regulated aphid density. Aphid density was highest in the cages, followed by open cages and lowest under no‐cage conditions. Numbers of D. noxia were generally higher compared to those of S. avenae. There was no significant difference in head length, mean kernel number/head, mean kernel mass/head or mean individual kernel mass between the two cultivars. Glutenin, gliadin and total protein content was analysed using size‐exclusion HPLC. Quality parameters of wheat flour were significantly different between the two cultivars. Gliadin content, total protein content and gliadin/glutenin ratio were significantly higher in Mv Magdaléna compared to Mv Magvas regardless of the level of caging and infestation, while glutenin content of uninfected Mv Magvas was significantly higher than that of Mv Magdaléna in the open cages. Aphid feeding did not induce any significant reduction in the gliadin, total protein content and gliadin/glutenin ratio of the examined hard wheat cultivars.CONCLUSION: Individual kernel mass reduction is the major factor in yield loss caused by aphids. Aphid feeding did not change the protein structure of hard‐kernel wheat. Copyright © 2007 Society of Chemical Industry

Influence of Neonicotinoid Insecticides on Infection by Neozygites fresenii (Nowakowski) Batko (Entomophthorales: Neozygitaceae) in the Cotton Aphid, Aphis gossypii Glover (Homoptera: Aphididae) in South Carolina

Incidence of infection by Neozygites fresenii (Nowakowski) Batko in the cotton aphid, Aphis gossypii Glover was monitored biweekly during the summer of 2002 in a cotton field in Bamberg County, SC. (33°22′02″N, 81°12′14″W). Five treatments were evaluated to determine the effects of insecticides on incidence of N. fresenii: acetamiprid, dicrotophos, thiamethoxam, imidacloprid, and control. Aphids were sampled by taking 24 leaves from 12 cotton plants from each plot and placed into 30-ml screw cap vials filled with 70% ethanol. Fungus infection in aphids, numbers of aphids, percentage of winged aphids, and fungus infection in winged aphids were determined from aphids for each plot.The highest fungus infection occurred on July 12 and 16. Acetamiprid treated plots had infection levels of N. fresenii lower than other treatments. Cotton aphid numbers in acetamiprid and thiamethoxam plots were significantly lower than in the dicrotophos treatment and untreated plots. None of the treatments caused reductions in percentage of winged aphids or in infection levels by N. fresenii in winged aphids.

Genetic variation in the effect of a facultative symbiont on host-plant use by pea aphids

Ecological specialisation on different host plants occurs frequently among phytophagous insects and is normally assumed to have a genetic basis. However, insects often carry microbial symbionts, which may play a role in the evolution of specialisation. The bacterium Regiella insecticola is a facultative symbiont of pea aphids (Acyrthosiphon pisum) where it is found most frequently in aphid clones feeding on Trifolium giving rise to the hypothesis that it may improve aphid performance on this plant. A study in which R. insecticola was eliminated from a single naturally infected aphid clone supported the hypothesis, but a second involving two aphid clones did not find the same effect. We created a series of new pea aphid-R. insecticola associations by injecting different strains of bacteria into five aphid clones uninfected by symbionts. For all aphid clones, the bacteria decreased the rate at which aphids accepted Vicia faba as a food plant and reduced performance on this plant. Their effect on aphids given Trifolium pratense was more complex: R. insecticola negatively affected acceptance by all aphid clones, had no effect on the performance of four aphid clones, but increased performance of a fifth, thus demonstrating genetic variation in the effect of R. insecticola on pea aphid host use. We discuss how these results may explain the distribution and frequency of this symbiont across different aphid populations.

Grass cultivar diversity and endophyte infection affect abundance of herbivores and their natural enemies

How does diversity in plant cultivars and endophyte infection affect higher trophic levels? We manipulated the number of cultivars (1 or 4) and the endophyte infection (-E, +E, and both, -E and +E) of potted Lolium perenne plants and left aphids and their parasitoids to assemble naturally. Aphid number and plant biomass were not influenced by our treatments, while the number of parasitised aphids (mummies) was significantly higher on mixed plant stands than on monocultures. The effect of endophytes was stronger in mixed plant stands than in monocultures with the most mummies found in endophyte-free mixed plant stands. Although number of mummies did not differ among cultivars, the rate of parasitism varied with cultivar and showed an endophyte x cultivar interaction. The number of successfully emerging parasitoids was also higher on high diversity treatments than on monocultures, indicating that increased diversity at resource levels translates to increased abundance at consumer levels. Key words: fungal endosymbionts, biodiversity, genetic diversity, multitrophic interactions, insect food webs, insect density, Neotyphodium lolii

Baking quality of wheat flour affected by cereal aphids

Flour from grains originating from plants infected artificially with cereal aphids were analyzed for glutenin and gliadin and total protein content, using Size Exclusion HPLC. Wheat plants were caged at the beginning of stem elongation. Cages were treated with 0.1 % methyl parathion. One week later, the caged plants were artificially infected with 5 aptera individuals of Metopolophium dirhodum, Diuraphis noxia, Sitobion avenae and Rhopalosiphum padi . It was found that aphid infection had significant effect on the glutenin and gliadin content, the total protein content and the gliadin/glutenin ratio. Both the glutenin and gliadin content was significantly higher in the seeds harvested from aphid infected plants. However, the gliadin/glutenin ratio was significantly lower in wheat flour prepared from aphid infected plants than in those from uninfected control. The most significant decrease in gliadin/glutenin ratio was caused by M. dirhodum, D. noxia, S. avenae infection followed by R. padi at high-abundan...

Cropping intensity and rainfall effects on upland rice yields in northern Laos

In northern Laos, upland rice is grown as a subsistence crop under rainfed conditions with no fertilizer inputs. It has traditionally been grown under slash-and-burn systems with long fallows, which restore soil fertility and reduce insect and weed pressure. However, increasing population density and government policies aimed at reducing the area under slash-and-burn have reduced fallows to as little as two or three years between rice crops. In this paper the impact of intensifying upland rice cultivation and rainfall on upland rice productivity was evaluated using yield and rainfall data from Luang Prabang province from 1992 to 2004. In addition, an experiment was conducted in 2004 to evaluate the effect of upland rice cropping intensification on soil nitrogen (N) and phosphate (P) availability and root pests (Tetraneura nigriabdominalis-root aphids and Meloidogyne graminicola Golden & Birchfield-nematodes). Rice yields were associated with total rainfall from June through August, corresponding to mid-tillering through flowering growth stages of upland rice. Increased cropping intensity resulted in a significant reduction of upland rice yields with higher rice yields being associated with longer fallows. Furthermore, when rice was annually cropped in the same field without fallows, rice yields rapidly declined. A study conducted in 2004 indicated that increasing cropping intensity reduced the soil N and P availability and increased root aphid infection of rice. The long-term productivity of upland rice can not be sustained with increased cropping intensity using the current management practices. Therefore, improved crop and resource management technologies are necessary for sustainable production.

Effect of Aphid Feeding on the Glutenin, Gliadin and Total Protein Contents of Wheat Flour

Plant sucking aphids cause both quantitative and qualitative yield losses in cereals; moreover aphid-transmitted viruses are responsible for other quantitative and qualitative damages, thus direct or indirect effects of aphid infection are in focus of interest. Bread-making quality of wheat flour is determined primarily by the protein content and composition, the gluten proteins (glutenins, gliadins) being the prime factors. Allelic composition of the gliadin- and glutenin loci as well as the absolute amount and/or the relative ratio of gliadins to glutenins are very important in dough making and in determining baking quality. Wheat plants were caged at the beginning of stem elongation. Cages were treated with 0.1% methyl parathion. One week later, the caged plants were artificially infected with 5 alata individuals of Metopolophium dirhodum, Diuraphis noxia, Sitobion avenae and Rhopalosiphum padi. Flour from grains originating from plants infected artificially with cereal aphids were analyzed for glutenin and gliadin and total protein content, using Size Exclusion HPLC. It was found that aphid infection had significant effect on the glutenin and gliadin content, the total protein content and the gliadin/glutenin ratio. Both the glutenin and gliadin content was significantly higher in the seeds harvested from aphid infected plants. However, the gliadin/glutenin ratio was significantly lower in wheat flour prepared from aphid infected plants than in those from uninfected control. The most significant decrease in gliadin/glutenin ratio was caused by M. dirhodum, D. noxia, S. avenae infection followed by R. padi at high-abundance and low-abundance, respectively. As the gliadin/glutenin ratio was significantly lower in flours made from aphid infected wheat seeds, it may be suggested that aphid feeding results in decreased bread making quality of wheat flour.

Effects of Hydrophilic and Hydrophobic Kaolin-Based Particle Films on Pea Aphid (Homoptera: Aphididae) and Its Entomopathogen <I>Pandora neoaphidis</I> (Entomophthorales: Entomophthoraceae)

Hydrophobic and hydrophilic kaolin-based particle films are effective for control of insect pests in certain agricultural crops. How these products interact with potential biological control agents is not well documented. This study was conducted to evaluate the effects of the hydrophobic (M96-018) and hydrophilic (Surround WP) kaolin-based particle films (Engelhard Corporation, Iselin, NJ) on pea aphid, Acyrthosiphon pisum (Harris), on peas (Pisum spp.), and on the fungal aphid pathogen Pandora neoaphidis (Remaudière and Hennebert) Humber. Over two field seasons (2001 and 2002) in northern Idaho, applications of M96-018 significantly reduced the rate of pea aphid increase on pea, but Surround WP, tested only in 2001, did not reduce aphid population growth rate. Neither particle film treatment was as effective as a standard application of esfenvalerate (DuPont Asana). In the laboratory, particle films suppressed pea aphid populations by up to 30%. M96-018 seemed to have some repellent activity based on aphid distributions after treating plants. When applied along with P. neoaphidis conidia, M96-018 but not Surround WP caused higher percentage of infection mortality of pea aphids by P. neoaphidis than occurred on controls treated only with P. neoaphidis conidia. P. neoaphidis conidia deposited on glass slides coated with M96-018, produced more germ tubes and secondary conidia than those deposited on untreated glass slides or slides treated with Surround WP. This result suggests that greater infection of pea aphids on plants treated with M96-018 is in part a result of a direct enhancement of fungal germination by the particle film.

Costs and benefits of symbiont infection in aphids: variation among symbionts and across temperatures.

Symbiosis is prevalent throughout the tree of life and has had a significant impact on the ecology and evolution of many bacteria and eukaryotes. The benevolence of symbiotic interactions often varies with the environment, and such variation is expected to play an important role in shaping the prevalence and distributions of symbiosis throughout nature. In this study, we examine how the fitness of aphids is influenced by infection with one of three maternally transmitted bacteria, 'Candidatus Serratia symbiotica', 'Candidatus Hamiltonella defensa' and 'Candidatus Regiella insecticola', addressing how symbiont benevolence varies with temperature. We find that the effects of these 'secondary' symbionts on Acyrthosiphon pisum depend on when and whether aphids are exposed to a brief period of heat shock. We also demonstrate that symbionts--even closely related isolates--vary in their effects on hosts. Our results indicate similar effects of S. symbiotica and H. defensa in conferring tolerance to high temperatures and a liability of R. insecticola under these same conditions. These findings reveal a role for heritable symbionts in the adaptation of aphids to their abiotic environments and add to an expanding body of knowledge on the adaptive significance of symbiosis.

Do aphids infected with entomopathogenic fungi continue to produce and respond to alarm pheromone?

The response of pea aphids, Acyrthosiphon pisum, to aphid alarm pheromone was not modified by infection with Beauveria bassiana. Approximately 50% of uninfected and infected aphids responded to synthetic alarm pheromone. The simulated attack of aphids infected with B. bassiana did not elicit a response in uninfected aphids. Preliminary air entrainment experiments of both uninfected aphids and aphids at different stages of B. bassiana (generalist pathogen) or P. neoaphidis (obligate pathogen of aphids) demonstrated that B. bassiana infected aphids produced less alarm pheromone than uninfected aphids and, conversely, P. neoaphidis infected aphids produced more alarm pheromone than uninfected aphids. These results are discussed with particular emphasis on the different life history strategies of these two pathogens. We hypothesise that the obligate, specialist pathogen, P. neoaphidis, is under greater selection pressure to increase pathogen transmission and survival resulting in modified host behaviour, than the generalist pathogen, B. bassiana.