Dispersal of Beauveria bassiana by the activity of nettle insects

Recent studies have shown that the entomopathogenic fungus Beauveria bassiana occurs naturally on the phylloplanes of several plants, including nettles. Insects could, by their activity, be contributing to this inoculum by dispersing it from other sites. The potential of nettle aphids Microlophium carnosum and their predator Anthocoris nemorum to disperse conidia of B. bassiana from soil to nettles and from sporulating cadavers in the nettle canopy was investigated in laboratory experiments. In petri dish assays, aphids showed potential to distribute B. bassiana from soil to nettle leaves. Predators dispersed inoculum from both soil and cadavers to nettle leaves in petri dishes. In microcosms, aphids did not disperse B. bassiana from the soil or from cadavers confined in the canopy, but A. nemorum were able to transfer inoculum from soil into the nettle canopy and to distribute conidia from cryptic cadavers. In some instances, infections were initiated in aphids and predators as a consequence of dispersal.     

Interactions Among the Aphid Diuraphis noxia , the Entomopathogenic Fungus Paecilomyces fumosoroseus and the Coccinellid Hippodamia convergens

Paecilomyces fumosoroseus (Wize) Brown & Smith is under development as a mycoinsecticide for control of the Russian wheat aphid, Diuraphis noxia Kurdjumov. Interactions with other natural enemies within the agro-ecosystem, such as the coccinellid Hippodamia convergens Guerin, require evaluation before its potential can be realized. In laboratory bioassays H. convergens adults were sprayed with suspensions of P. fumosoroseus conidia at different concentrations (including potential field rates) and mortality assessed. Although a proportion of coccinellids succumbed to infection (the greatest proportion was 22% when the ladybirds had suffered stress) it is unlikely that they would be at risk from infection as a direct result of a spray application unless there were prolonged periods of high humidity or the coccinellids were stressed. When provided with uninfected or P. fumosoroseus -infected D. noxia cadavers as prey, coccinellids consumed more uninfected aphids. The predators never consumed aphid cadavers from which the fungus was erupting or sporulating. Hippodamia convergens is, therefore, unlikely to be a significant intraguild predator of P. fumosoroseus . Predators contaminated with conidia of P. fumosoroseus using different methods (sprayed coccinellids, coccinellids foraging on sprayed aphids, and those foraging in the presence of sporulating D. noxia cadavers) were able to transfer conidia to healthy D. noxia populations and initiate infection in a proportion of those aphids. The proportion of aphids becoming infected was greatest when the coccinellids became contaminated when foraging amongst sporulating cadavers. Some coccinellids also succumbed to infection under these conditions.     

Effect of seasonal abiotic conditions and field margin habitat on the activity of Pandora neoaphidis inoculum on soil

The ability of the aphid pathogenic fungus Pandora neoaphidis to remain active in the absence of a resting stage through a combination of continuous infection and as conidia deposited on soil was assessed alongside the potential for planted field margins to act as a refuge for the fungus. P. neoaphidis was able to infect the pea aphid, Acyrthosiphon pisum, when maintained under controlled conditions that simulated those that occur seasonally in the UK. Although there was a significant inverse relationship between temperature and time-to-kill, with death occurring after 4.2, 6.9 and 13.6 days when maintained under fluctuating summer, autumn and winter temperatures, respectively, there were no additional statistically significant effects of photoperiod. The activity of inoculum on soil was indirectly assessed by baiting with A. pisum. Under controlled conditions P. neoaphidis remained active on soil and was able to infect aphids for up to 80 days. However, the percentage of aphids that became infected decreased from 76% on day 1 to 11% on day 80. Whereas there was little difference in the activity of conidia that had been maintained at 4 degrees C and 10 degrees C, activity at 18 degrees C was considerably reduced. Under field conditions the activity of inoculum was strongly influenced by season. On day 49 there was little or no activity during spring, summer or winter. However, during autumn a mean proportion of 0.08 aphids still became infected with P. neoaphidis. Margin type did not affect the activity of conidia nor was there a difference in activity between blocks that had regenerated naturally and those that had been planted. These results suggest that P. neoaphidis can infect aphids and remain active on soil under the abiotic conditions that occur seasonally in the UK and that this fungus may be able to persist annually without a resting stage.     

Cage and field assessments of Beauveria bassiana-based Mycoinsecticides for Myzus persicae Sulzer (Hemiptera: Aphididae) control in cabbage

The efficiency of formulated Beauveria bassiana-based mycoinsecticides to control Myzus persicae (Sulzer) in cabbage was assessed under field conditions. Aqueous conidial suspensions (0.01% Tween 80 + 0.01% v/v Agral) of three fungal isolates were sprayed twice at different dates, each with 2.0 x 10(9) viable conidia per potted plant using screened cages. The number of nymphs and adults of M. persicae per leaf was significantly reduced in plots treated with isolates CG 864 and PL 63, with control efficiency ranging from 57% to 60%. Further field trials using screened cages with isolate CG 864 formulated as oil dispersion reduced the aphid population by 85-87% as compared to the control, whereas a 71% reduction was seen in plants treated with the aqueous conidial suspension 20 days following the first spray. The last experiment was conducted in a commercial cabbage field (without cages), in which the fungus was applied at three different dates, each with an equivalent of 1.0 x 10(13) viable conidia/ha. The reduction in the number of aphids per leaf was more evident between four and five weeks following the first spray, resulting in 76-83% and 57-65% control efficiency for oil dispersions and unformulated conidia, respectively. However, with the exception of imidacloprid-treated plants, rapid aphid re-infestation was observed in all treatments. In this study, the stand-alone use of mycoinsecticides for aphid control was not a satisfactory strategy, although utilization of B. bassiana in IPM strategies remains a field to be explored.     

Effects of hydrophilic and hydrophobic kaolin-based particle films on pea aphid (Homoptera: Aphididae) and its entomopathogen Pandora neoaphidis (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.     

Plant waxy bloom on peas affects infection of pea aphids by Pandora neoaphidis

This study examined the effects of the surface wax bloom of pea plants, Pisum sativum, on infection of pea aphids, Acyrthosiphon pisum, by the fungal pathogen Pandora neoaphidis. In prior field surveys, a higher proportion of P. neoaphidis-killed pea aphids (cadavers) had been observed on a pea line with reduced wax bloom, as compared with a sister line with normal surface wax bloom. Laboratory bioassays were conducted in order to examine the mechanisms. After plants of each line infested with aphids were exposed to similar densities of conidia, the rate of accumulation of cadavers on the reduced wax line was significantly greater than on the normal wax bloom line; at the end of the experiment (13d), the proportion of aphid cadavers on the reduced wax line was approximately four times that on the normal wax bloom line. When plants were exposed to conidia first and then infested with aphids, the rate of accumulation of cadavers was slightly but significantly greater on the reduced wax line, and infection at the end of the experiment (16d) did not differ between the lines. When aphids were exposed first and then released onto the plants, no differences in the proportion of aphid cadavers were observed between the pea lines. Greater infection of pea aphid on reduced wax peas appears to depend upon plants being exposed to inoculum while aphids are settled in typical feeding positions on the plant. Additional experiments demonstrated increased adhesion and germination by P. neoaphidis conidia to leaf surfaces of the reduced wax line as compared with normal wax line, and this could help explain the higher infection rate by P. neoaphidis on the reduced wax line. In bioassays using surface waxes extracted from the two lines, there was no effect of wax source on germination of P. neoaphidis conidia.     

球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响

在室内评价了球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响。分别在烟蚜茧蜂寄生桃蚜后不同时间进行高剂量(1900孢子/mm2)接菌,检测蚜虫感病率和寄生蜂形成的僵蚜率及僵蚜出蜂率。结果表明,球孢白僵菌对僵蚜率和僵蚜出蜂率的影响随接菌时间不同而变化。在烟蚜茧蜂寄生前1d、寄生当天和寄生后3d接菌,蚜虫感病率分别为59.6%、56.2%和34.8%;与对照相比,僵蚜率分别下降94%、59%和47%,僵蚜出蜂率分别减少83%、54%和49%。在寄生后5d或7d接菌,僵蚜率和僵蚜出蜂率不受明显影响,但蚜虫感病率降低到8.2%以下。对蚜尸内白僵菌菌体含量检测表明,随着烟蚜茧蜂寄生后接菌时间的推移,菌体数量迅速下降。寄生蜂寄生后5d或7d接菌,蚜尸内几乎检测不到菌体。直接喷雾接菌烟蚜茧蜂,成蜂寿命缩短4d左右,且81.8%的蜂尸受白僵菌感染。接菌后的寄生蜂对蚜虫寄生率几乎无影响,但寄生蜂在蚜虫体内的存活时间缩短了27.8%。     

Effect of fungal infection on the reproductive potential of aphids and their progeny

The effect of infection by Pandora neoaphidis and Beauveria bassiana on the reproductive potential of the pea aphid, Acyrthosiphon pisum, and their progeny was assessed. Infection by either P. neoaphidis or B. bassiana reduced the number of nymphs produced within 24 h of inoculation and over the entire infection period compared to uninfected aphids. However, infection by either P. neoaphidis or B. bassiana for 24 or 72 h did not alter the intrinsic rate of increase of the host aphid's progeny. Therefore, fungal infection appears to have no indirect effects on the fitness of the host's progeny.     

Efficacy of Bauveria bassiana for red flour beetle when applied with plant essential oils or in mineral oil and organosilicone carriers

The carriers mineral oil and Silwet L-77 and the botanical insecticides Neemix 4.5 and Hexacide were evaluated for their impacts on the efficacy of Beauveria bassiana (Balsamo) Vuillemin conidia against red flour beetle, Tribolium castaneum (Herbst), larvae. The dosages of liquid treatments were quantified by both conidia concentration in the spray volume and conidia deposition on the target surface. The latter approach allowed comparison with dry, unformulated conidia. The median lethal concentrations of B. bassiana in 0.05% Silwet L-77 solution or without a carrier were approximately double that for conidia in mineral oil. Carriers had highly significant effects on the efficacy of B. bassiana. The lower efficacy of conidia in aqueous Silwet L-77 may have been the result of conidia loss from the larval surface because of the siloxane's spreading properties. Neemix 4.5 (4.5% azadirachtin) delayed pupation and did not reduce the germination rate of B. bassiana conidia, but it significantly reduced T. castaneum mortality at two of four tested fungus doses. Hexacide (5% rosemary oil) caused significant mortality when applied without B. bassiana, but it did not affect pupation, the germination rate of conidia, or T. castaneum mortality when used in combination with the fungus.     

Influence of Experience on the Response of Bathyplectes curculionis (Hymenoptera: Ichneumonidae), a Nonaphidophagous Parasitoid, to Aphid Odor

Bathyplectes curculionis (Thomson) is an introduced natural enemy of the alfalfa weevil in North America. The wasp requires carbohydrate foods as an adult. Adult wasps have increased longevity and fecundity when provided access to pea aphid, Acyrthosiphon pisum (Harris), honeydew in the laboratory, and adults respond positively to the presence of pea aphids in alfalfa fields. However, it is unknown how these wasps find aphid honeydew in the field. In a series of Y-tube olfactometer experiments, we evaluated the response of naïve and experienced adult female B. curculionis to odors from pea aphids, alfalfa, and pea aphids on alfalfa. Naïve adult females did not respond positively to pea aphid odor even when hungry. But adult females were able to learn aphid odor, and the mechanism of learning appears to be associative rather than by sensitization. Naïve females also showed no preference for alfalfa odor but learned alfalfa odor through sensitization. The wasps did not distinguish between alfalfa with aphids and alfalfa without aphids, even after exposure to aphids or alfalfa with aphids. However, they preferred pea aphid odor to alfalfa odor after a feeding experience in the presence of pea aphid odors. But after exposure to mixed odors of aphids and alfalfa while feeding, B. curculionis females preferred the odor of alfalfa to the odor of pea aphids. These results suggest that alfalfa odors mask or override aphid odors when aphids are associated with alfalfa (as happens naturally), thus interfering with the wasp's ability to respond to learned aphid odors. Therefore, although the wasps are capable of learning to find pea aphids and their honeydew in a simplified laboratory setting, it appears unlikely that they do so in the field.     

Spatial Simulation of Epizootics Caused by Beauveria bassiana in Russian Wheat Aphid Populations

Models of disease dynamics commonly make the assumption of spatial homogeneity in the underlying host population. However, insect behavior may result in spatially heterogeneous populations with which pathogens interact. We modified a simulation model of temporal and spatial population dynamics of the Russian wheat aphid, Diuraphis noxia, on preferred or nonpreferred host plants, by incorporating effects of the entomopathogenic fungus, Beauveria bassiana. Epizootic parameters included time from inoculation of aphids until death, duration of sporulation, and estimated exposure probability. Simulations first predicted results of previously described experiments in which D. noxia adults were inoculated with conidial suspensions or water and placed on wheat or oat seedlings in 81-plant grids in cages. Subsequently, large-scale simulations were run for hypothetical field situations on 50 × 50-plant grids of wheat or oat. With B. bassiana present for both cage and larger scale simulations, results indicated that, on oat, an expanding infection front lagged behind the expanding aphid population front. Continual aphid movement from hosts resulted in many escapes, and the aphid population persisted at slightly reduced levels. On the preferred wheat host, patterns developed with pockets of infected aphids and other pockets of healthy aphids. Localized aphid populations that escaped initial infestation were able to proliferate, whereas other local populations were greatly reduced or became extinct due to lack of movement from the hosts, resulting in increased exposure to pathogen inoculum. Thus, proliferation and fluctuation of the pathogen were strongly influenced by the plant hosts' effects on aphid movement behavior. Incorporating spatial dynamics into disease models should prove useful in other efforts to predict biological control efficacy by entomopathogenic fungi in heterogeneous habitats.     

The persistence of infectivity of conidia of the aphid pathogen Erynia neophidis on leaves in the field

Conidia of the aphid pathogen Erynia neophidis inoculated on to bean leaves in the field in 1980, 1981 and 1982, retained some infectivity for pea aphids for at least 14 days. Decline in infectivity occurred linearly with time. Inocula on leaves near the base of plants remained infective longer than those on leaves near the top and those on the abaxial surface remained infective for longer than those on the adaxial surface. Infectivity persisted longest in 1981 when conditions were coolest with least sun.     

Impact of the entomopathogenic fungus Verticillium lecanii on development of an aphid parasitoid, Aphidius colemani

The parasitoid Aphidius colemani developed normally (approximately 90% adult emergence) when its cotton aphid (Aphis gossypii) host was treated with Verticillum lecanii conidia 5 or 7 days after parasitization. Fungus exposure 1 day before or up to 3 days after parasitization, however, reduced A. colemani emergence from 0 to 10%. Also, numbers of spores and mycelial fragments in aphid homogenates were much higher in aphids exposed to the fungus up to 3 days after parasitization than in aphids treated after 5 or 7 days. Our results suggest that the parasitoid and fungus may be used together for aphid biocontrol as long as fungus applications are timed to allow late-instar development of the parasitoid.     

Laboratory observations on the effect of humidity on the persistence of infectivity of conidia of the aphid pathogen Erynia neoaphidis

Conidia of the aphid pathogen Erynia neoaphidis on detached bean leaves and glass coverslips maintained at 20°C and at humidities from 40 to 100% r.h. lost their infectivity for pea aphids at a rate dependent on the humidity. Infectivity declined most rapidly in inocula kept on leaves at 70% r.h. and persisted longest in those at 40 and 50% r. h. That of inocula on coverslips declined most rapidly at 77% r.h. and persisted longest in those kept at 40% r.h. Even after 21 days a little infectivity was retained by conidia stored at 50% r.h. on leaves and 40% r.h. on coverslips.     

球孢白僵菌对桃蚜接种后特定时间内的侵染率

用球孢白僵菌 (Beauveriabassiana)BBSG870 2菌株的分生孢子悬液 (5× 10 6个·ml-1孢子 )对桃蚜 (Myzuspersicae)始产若蚜的成蚜进行表面接种 ,在接种后 5 6h内 ,每隔 8h取样用 0 .2 %百菌清处理蚜虫 ,使其体表残存的孢子全部失活 ,分别置于 10℃和 2 0℃下逐日观察感染引起的死亡 .结果表明 ,10℃下接种后 5 6h、2 0℃下接种后 40h内各时间段有效侵染引起的死亡率相互间存在显著差异 ,并与不用杀菌剂的对照处理差异显著 (P <0 .0 5 ) .与对照相比 ,接种后 8、16、2 4、32、40和 5 6h内 ,10℃下的有效侵染率分别为 2 2 .9、48.8、6 4.9、80 .4、72 .7和 98.3% ,2 0℃下分别为 31.6、48.8、5 8.6、86 .9、97.2和 98.7% .由此表明 ,在 10～ 2 0℃范围内 ,接种后 2 4h内是该菌有效侵染桃蚜的关键时段 ,有效侵染率达 5 9～ 6 5 % .     

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

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

Pathogenicity of hyphomycet fungi to aphids Aphis gossypii Glover and Myzus persicae (Sulzer) (Hemiptera: Aphididae)

The aphids Aphis gossypii and Myzus persicae are cosmopolitan, poliphagous and damage cultivated plants. The effects of the entomopathogenic fungi Beauveria bassiana (isolate IBCB 66), Metarhizium anisopliae (isolate IBCB 121), Paecilomyces fumosoroseus (isolate IBCB 141) and Lecanicillium (= Verticillium) lecanii (isolate JAB 02) on third instar nymphs of A. gossypii and M. persicae were evaluated in the laboratory at 25 degrees C, 70 +/- 10% RH and 12h photophase. The aphids were transferred to petri dishes with a foliar disk (cotton or pepper) with a layer of 1 cm tick of agar-water. The fungi were applied in a suspension containing 1.0 x 106 to 1.0 x 108 conidia/ml. In the control treatment 1 ml of sterilized water was added to the foliar disks. The mortality of aphids was evaluated daily. B. bassiana and M. anisopliae caused 100% mortality at the seventh day after inoculation, for both species. L. lecanii was the fungus that provided mortality later in the aphids and M. persicae was more susceptible to both fungi than A. gossypii.     

Identification of two clip domain serine proteases involved in the pea aphid's defense against bacterial and fungal infection

Phenoloxidases (POs) are required for the pea aphid's defense against bacterial and fungal infection. Prophenoloxidases (PPOs) are proteolytically converted to its active form PO through a clip domain serine protease cascade. In this study, we identified five clip domain serine proteases in the pea aphids. The messenger RNA levels of two of them, Ap_SPLP and Ap_VP, were upregulated by Gram‐positive bacterium Staphylococcus aureus and fungus Beauveria bassiana infections. Double‐stranded RNA‐based expression knockdown of these two genes resulted in reduced PO activity of the aphid hemolymph, higher loads of S. aureus and B. bassiana in the aphids, and lower survival rates of the aphids after infections. Our data suggest that Ap_SPLP and Ap_VP are involved in PPO activation pathway in the pea aphid.     

A plant surface mutation mediates predator interference among ladybird larvae

Abstract.  1. We investigated mechanisms causing predator–predator interference between fourth instar Hippodamia convergens larvae foraging for pea aphids on pea plants, Pisum sativum , with a wild-type wax bloom, and the lack of such interference between larvae foraging on pea plants with a reduced-wax bloom caused by the single gene mutation wel .
2. Observations showed that behavioural interactions between larvae were not affected by wax phenotype. Specifically, larvae did not encounter one another more frequently on normal-wax peas as may have been predicted because reduced ability by coccinellids to attach to normal-wax plant surfaces could restrict them to foraging on only some parts of these plants.
3. In a controlled bioassay on normal-wax peas, H. convergens larvae avoided leaflets previously exposed to another larva. On reduced-wax peas, this effect was not detected.
4. In microcosm experiments, inter-predator interference in terms of prey consumption occurred on normal-wax peas, but not on reduced-wax peas. The interference on normal-wax peas occurred whether two H. convergens larvae were placed on a pea aphid-infested, normal-wax plant simultaneously or sequentially.
5. We conclude that the observed inter-predator interference is not as a result of direct physical contact, but rather arises because of (i) inhibition of foraging by chemical trails left by other larvae, (ii) the inability of larvae to access portions of the normal-wax plants creating aphid refugia, or (iii) a combination of these factors.     

Shoot-feeding Aphids Promote Development of Gremmeniella abietina, the Fungal Pathogen Causing Scleroderris Canker Disease in Conifers

We tested experimentally whether shoot feeding aphids Cinara pinea (Mordv.) can promote the development of Gremmeniella abietina (Lagerb.) Morelet, a fungus which causes Scleroderris canker disease in conifers. Pine seedlings were infested with aphids at two different times, and subsequently inoculated with conidia of G. abietina at two different times. The degree of infestation was classified into three groups based on the number of aphids/seedling: none, one (low level), and three (high level). Because of parthenogenetic reproduction, the number of aphids increased during the summer, so we also used aphid numbers to explain the development of the symptoms. Necrosis caused by G. abietina was more prevalent in seedlings infested by aphids in June, and inoculated with fungus in July. Expressed as percents of the total length of the shoots, the necrosis had advanced 50%, 70%, and 95% with no, low, and high aphid levels, respectively. Canker formation was significantly greater when G. abietina conidia were inoculated in July, compared to August. In August, there were no statistically significant differences in the intensity of disease between aphid infestation levels, but the mean number of aphids was related to disease intensity. In the spring following infection, seedlings with aphids had more dead and fewer healthy terminal buds compared to seedlings without aphids.