Pathogenicity of six isolates of Beauveria bassiana (Balsamo) Vuillemin (Deuteromycotina, Hyphomycetes) to Perillus bioculatus (F) (Hem., Pentatomidae)

Abstract:  The two-spotted stink bug, Perillus bioculatus (F) (Hem., Pentatomidae) is considered an important predator of Colorado potato beetle egg masses and small larvae. The susceptibility of P. bioculatus second instar nymphs to six isolates of Beauveria bassiana (Balsamo) Vuillemin, which are highly pathogenic to Colorado potato beetle, was tested in the laboratory. Five isolates were highly pathogenic and caused more than 77% mortality after 8 days at a concentration of 10⁶ conidia/ml. However, isolate IPP46 showed low pathogenicity and caused only 11% mortality of nymphs. The pathogenicity of isolates was independent of their host, source or country of origin. Isolate IPP46 was tested at six different concentrations. A linear relationship between the concentration of B. bassiana and the mortality of nymphs was observed. The 50% lethal concentration (LC₅₀) value was 3.4 × 10⁷ conidia/ml. The LT₅₀ decreased with increase of conidia concentration used. The present results suggest that isolate IPP46 has good potential as a biological control agent within an integrated pest management programme.     

Bacillus thuringiensis var. tenebrionis shows no toxicity to the predator Chauliognathus lugubris (F.) (Coleoptera: Cantharidae)

Laboratory trials examined the effect of the bioinsecticide Novodor^® (active ingredient 3% Bacillus thuringiensis var. tenebrionis (w/w) (Btt)), on the Australian plague soldier beetle, Chauliognathus lugubris, a predator of the Tasmanian eucalyptus leaf beetle Chrysophtharta bimaculata . Spraying of C. lugubris and the C. bimaculata eggs supplied as food showed that neither Novodor^® FC with active nor inactive Btt caused any decrease in predatory performance, longevity or fecundity of C. lugubris during the period of their population peak (December). Towards the end of their peak population levels, in late January, there was a steady increase in mortality which was not related to either of the Novodor^® FC treatments. On average, male:female pairs of C. lugubris laid one egg batch and ate 30 C. bimaculata eggs a week whether untreated or sprayed with either Novodor^® FC. This Btt-based insecticide appears to be a suitable component of an integrated pest management (IPM) regime that aims to conserve natural predators.     

Respiration rhythms and heartbeats of diapausing Colorado potato beetles, Leptinotarsa decemlineata, at low temperatures

Discontinuous gas exchange cycles (DGCs), active muscular ventilation, microcycles of repetitive openings, and heartbeats of diapausing adult Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae), were studied at low temperatures (0, 5, and 10 °C) using an electrolytic respirometer combined with an infrared actograph. The DGC of the adult constriction-flutter-open type was the main respiration mode in fully quiescent beetles at temperatures from 5 to 10 °C. The CO₂ bursts were actively ventilated at temperatures above 5 °C. During the flutter period, a series of microcycles appeared, but no muscular contractions associated with the microcycles were detected. We identified this respiration mode as discontinuous suction ventilation.
The hydration condition of the beetles did not influence the frequency of the gas exchange cycles, but dehydrated beetles showed significantly longer flutter periods and shorter ventilation periods than hydrated beetles. The heartbeat frequencies were influenced by both temperature and hydration status.
We conclude from the results that DGCs are used at rest in adult L. decemlineata under various environmental conditions and also at low temperatures. Our results showed that DGCs are the main respiration mode of resting adult Colorado potato beetle irrespective of its hydration state and temperature. Our method resolves O₂ uptake and subsequent CO₂ release in flutter and ventilation periods and shows that diffusion is replaced by convection to reduce water loss in adult beetles.     

Efficacy of Beauveria bassiana (Bals.) Vuill. against the spruce bark beetle, Ips typographus L., in the laboratory under various conditions

Abstract:  In laboratory bioassays, the efficacy of the entomopathogenic fungus Beauveria bassiana against the spruce bark beetle, Ips typographus , was tested under various conditions. Four of the tested isolates and the commercial product Boverol^® caused 99–100% mortality when tested at a concentration of 1.0 × 10⁷ conidia/ml at 25°C. Using B. bassiana isolate 138 at a concentration of 1.0 × 10⁶, the median survival time (MST) was 6.1 d and significantly longer compared with the MST of 4.2 and 4.0 d at 1.0 × 10⁷ and 1.0 × 10⁸ conidia/ml, respectively. In the next experiment, the beetles were maintained on spruce bark, filter paper or artificial diet during the bioassay with Boverol^®, and significant differences in the MST of 3.6, 2.5 and 5.3 d, respectively, were noticed. The experiment with Boverol^® at different temperatures showed that the beetles lived significantly longer at 15°C (MST 8.7 d) than at 20, 25, 30 and 35°C. At 25°C, the beetles died most rapidly (MST 3.5 d). At different relative humidities (RH) of 40, 70 and 100%, nearly all beetles were dead after treatment with a suspension of Boverol^® at 1.0 × 10⁷ conidia/ml. At 40% RH, 49% of the untreated beetles died after 7 d. The best effects were achieved with the following bioassay: beetles were fed for three days on artificial diet, then dipped into a solution of 1.0 × 10⁷ conidia/ml and transferred on a piece of spruce bark in Petri dishes at 25°C and 70% RH.     

Heat‐induced mortality and expression of heat shock proteins in Colorado potato beetles treated with imidacloprid

The Colorado potato beetle is an important pest of solanaceous plants in the Northern Hemisphere. Better understanding of its physiological responses to temperature stress and their interactions with still‐prevalent chemical control has important implications for the management of this insect. We measured mortality and expression of the Hsp70 heat shock proteins in the Colorado potato beetle larvae exposed to sublethal concentration of the commonly used insecticide imidacloprid, and to supraoptimal temperatures. Both turned out to be significant stress factors, although induction of Hsp70 by imidacloprid observed in the present study was low compared to its induction by the heat. The two factors also interacted with each other. At an extreme temperature of 43 °C, exposure to a sublethal dose of imidacloprid resulted in a significant rise in larval mortality, which was not observed at an optimal temperature of 25 °C. Heat‐stressed larvae also failed to respond to imidacloprid by producing more Hsp70. These findings suggest that when field rates of insecticides become insufficient for killing the exposed beetles under optimal temperature conditions due to the evolution of resistance in beetle populations, they may still reduce the probability of resistant beetles surviving the heat shock created by using propane flamers as a rescue treatment.     

Pymetrozine causes a nontarget pest, the Colorado potato beetle (Coleoptera: Chrysomelidae), to leave potato plants

Pymetrozine is a selective insecticide that targets aphids. Published assessments of the effects of pymetrozine on nontarget organisms focus mainly on predatory insects, and they rarely indicate toxicity. In a laboratory bioassay, survival of Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), larvae was not affected by pymetrozine exposure. We subsequently used pymetrozine to implement low-aphid-density treatments in a field experiment that involved separate manipulations of Colorado potato beetle density. Unexpectedly, the addition of Colorado potato beetle adults and eggs did not increase the densities of Colorado potato beetle larvae in plots that were sprayed with pymetrozine (applied with water and an adjuvant). In control plots sprayed with water and adjuvant (without pymetrozine), addition of Colorado potato beetles increased densities of their larvae. Data collected on a smaller scale suggest that a behavioral mechanism underlies the population-level pattern: Colorado potato beetle larvae become more active and are less likely to remain on a host plant after exposure to pymetrozine. Thus, potato, Solanum tuberosum L., growers who use pymetrozine against aphids also might benefit in terms of Colorado potato beetle control.     

A model for determination of the numbers of pollen beetles (Meligethes aeneus F.) (Col., Nitidulidae) per plant in oil-seed rape crops (Brassica napus L.) by estimating the percentage of plants attacked by pollen beetles

Abstract:  Pollen beetles ( Meligethes aeneus F) are common pests of oil-seed rape crops ( Brassica napus L) in Denmark, causing serious yield losses (>80% reduction). In consequence, the crop is treated with pesticides in most years. To estimate the need for insecticide treatment it is desirable to develop a method, which, in a simple and very labour-saving way, can estimate the number of pollen beetles in oil-seed rape crops. Incidences of pollen beetles were recorded in 105 spring and winter oil-seed rape fields. From the coherent values of the percentage of plants infested with pollen beetles ( Y ) and the number of pollen beetles per plant ( X ) the following model was deduced: Y  = 43.3 ×  X ^0.47.     

The role of sulphur compounds for breeding success of Ips typographus L. (Col., Scolytidae) on Norway Spruce (Picea abies [L.] Karst.)

Bark beetles, especially Ips typographus L. represent a severe biotic threat for spruce ( Picea abies [L.] Karst.) at low altitudes in Europe. We compared sulphur (total S, SO₄²⁻, glutathione, cysteine, methionine), nitrogen (total N, NO₃⁻, total protein, free amino acids), carbon, total phosphorus and PO₄³⁻, tree vigour index (TVI) and water content of the phloem after felling, and their dependent changes (tdc) with the breeding success of I. typographus . Twenty trees were classified according to age (34/90 years) and crown density (high/intermediate/low). Water content was higher in young trees than in old trees, higher in the crown than at breast height, and decreased significantly within the 8-week study period. In old trees, breeding success, length of mother galleries and SO₄²⁻ were significantly higher, while total protein, NO₃⁻ and water content were significantly lower than in young trees. Trees with intermediate crown density provided the best breeding success for I. typographus and had significantly higher arginine content and C/N ratio as well as low amounts of phosphate and glutamine. During the period of bark beetle breeding, total sulphur, glutathione, protein, NO₃⁻, aspartate, glutamine, glutamate, arginine and γ-aminobutyrate decreased significantly. The results support previous investigations that I. typographus develops best in physiologically weakened trees.     

Field efficacy of the entomopathogenic nematode Steinernema feltiae against the Mediterranean flat-headed rootborer Capnodis tenebrionis

The Mediterranean flat-headed rootborer, Capnodis tenebrionis (L.) (Coleoptera: Buprestidae), is an economically important pest of stone fruit and seed fruit in Mediterranean areas. The potential control of the entomopathogenic nematode Steinernema feltiae (Filipjev) (strain Bpa), isolated from a dead C. tenebrionis larva, was tested in a cherry tree orchard in Ullastrell, Barcelona (Spain). Nematode infective juveniles (IJs) were applied by drench and injection. In both the treatments, a rate of 1 million IJs was applied per tree every week during 4 or 8 weeks, with a total dose of 4 × 10⁶ IJs/tree and 8 × 10⁶ IJs/tree. Number, stage and localization of insects in each tree trunk were recorded. In both the experiments, S. feltiae significantly reduced the population of C. tenebrionis providing control ranging from 88.3% to 97%. No significant differences were recorded between the different treatments. Persistence of nematodes was recorded until 6 weeks after application. Results indicate that the application of S. feltiae (Bpa) provides adequate control of C. tenebrionis in cherry trees.     

Use of sperm precedence to infer the overwintering cost of insecticide resistance in the Colorado potato beetle

1 Resistance to insecticides is a model system for studying adaptation. Although selection for resistance is always strong in areas and seasons where populations are exposed to insecticides, costs of resistance, which may only be expressed in the absence of insecticide use, will shape how quickly resistance will evolve.
2 We used sperm precedence to measure the shifts in resistance to imidacloprid in a natural population during winter diapause in the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). Because adult beetles overwinter with viable autumn sperm, but sperm from spring matings take precedence over stored sperm, we used the difference in resistance of springmated and autumn-mated overwintered females to estimate the shift in resistance during the winter. Offspring of autumn-mated females were 2.7- or 2.5-fold more resistant compared with offspring from spring-mated females in two replicate trials.
3 We also measured the resistance of late summer and spring emergent adults in fields treated and untreated with imidacloprid in the first year. Adults from the treated field were 13.7-fold more resistant and adults from the untreated field were 2.6-fold more resistant compared with the next spring's emergers.
4 These large costs of resistance observed in the field and inferred from resistance declines during diapause help to explain the observation that imidacloprid resistance has increased only slowly over the decade of widespread use against this species, and how insecticide resistance in general can cycle annually.     

Impact of Bacillus thuringiensis – insecticides on population dynamics and egg predation of the Colorado potato beetle in North Carolina potato plantings

Field studies to assess the impact of Bacillus thuringiensis var. tenebrionis (Btt)-insecticides on Colorado potato beetle populations, egg survivorship and levels of predation on egg masses were conducted in replicated field research plots during two years. Stage-specific abundance of the Colorado potato beetle and predation on egg masses were monitored in Btt-treated and untreated potato plots in both years. The Btt-treatments significantly reduced densities of large (third and fourth instar) Colorado potato beetle larvae. The densities of large larvae remained below 0.5 and 3 per plant in the Btt-treatment while peak densities of 4.5 and 21 large larvae per plant occurred in the untreated control in 1992 and 1993, respectively. Regular sampling of egg masses indicated that predation rates in Btt-treated and untreated plots did not differ significantly although, in 1993, predation rates of up to 100% were recorded, only in Btt-treated plots. In a predator exclusion study carried out in 1992, survivorship of protected eggs was consistently higher than of eggs exposed to predation. Seasonal survivorship of exposed eggs was significantly lower in the Btt-treated than in untreated plots. Btt insecticides for control of Colorado potato beetles provided direct protection of the crop and were compatible with naturally-occurring biological control of Colorado potato beetle eggs due to predation.     

Effects of toxins from two strains of Verticillium lecanii (Hyphomycetes) on bioattributes of a predatory ladybeetle, Delphastus catalinae (Col., Coccinellidae)

Abstract:  The aim of this study was to test the hypothesis that the whitefly ( Bemisia tabaci ; Hom., Aleyrodidae) predator ladybird beetles, Delphastus catalinae (Col., Coccinellidae), are not adversely affected in the field by the crude insecticidal toxins extracted from two strains of the fungus Verticillium lecanii , V3450 and Vp28. We developed a method to evaluate sublethal toxicity and its effects on consumption and functional response of D. catalinae . The crude toxins have low toxicity against beetle larva with LC₅₀ values of 1942 (1393–2710) and 2471 (1291–4731) p.p.m., respectively (approximately 10- and 12-fold of field rate of application 200 p.p.m.). The adult beetles had less sensitivity to crude toxins with LC₅₀ values of 4260 (3376–5375) and 4426 (1734–11298) p.p.m., respectively (approximately 20- and 22-fold of field rate 200 p.p.m.). The consumption and foraging capacity were significantly impaired especially in the second-instar larval beetles which took longer time (more than twice of the control beetles) to consume whitefly eggs after exposure to toxins, although D. catalinae suffered no significant effect on fecundity and longevity, when exposed to a toxin dilution of field rate. The data suggest that spraying of V. lecanii or its toxins should be avoided in the field having immature stages of D. catalinae .     

Behavioural responses of the Colorado potato beetle to trichomes and leaf surface chemicals of Solanum tarijense

Wild Solanum species constitute a source of resistance to several pests and diseases of potato. Several species of wild tuber‐bearing potato have been identified as resistant to the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), including Solanum tarijense Hawkes (Solanaceae). Our objective was to determine the mechanism of resistance of S. tarijense to the Colorado potato beetle and, because the resistance is limited to the adult stage of the insect, to study the host selection behaviour on resistant plants. In the field, Colorado potato beetles demonstrated a unique behaviour when in contact with S. tarijense, abandoning the plant by falling to the ground after a few minutes. The abundant trichomes on the leaves of S. tarijense induced the falling behaviour. However, on S. tarijense feeding remained low even after the trichomes were mechanically removed. Observations demonstrated that the normal sequence of behaviour leading to feeding was interrupted before adult beetles fed on S. tarijense leaves. Feeding experiments using volatile and non‐volatile fractions of leaf surface extracts identified a phagodeterrent effect of the volatile fraction. Our results contrast with a similar evaluation of the mode of resistance of Solanum berthaultii Hawkes, a close relative of S. tarijense, on which some feeding occurred and adults did not show falling behaviour. This study presents information on S. tarijense as a new source of resistance to the Colorado potato beetle that can be used for potato breeding.     

Baseline toxicity and field efficacy of metaflumizone on Colorado potato beetle (Coleoptera: Chrysomelidae)

Baseline toxicity levels to a novel semicarbazone insecticide, metaflumizone were established for 25 field populations of Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae),from North America. Excluding the susceptible laboratory strain, 50% lethal concentrations of metaflumizone ranged from 0.57 to 1.31 ppm, while response slopes ranged from 1.92 to 4.24 (average = 2.93), and were unrelated to the 50% lethal concentration (r = 0.06; P = 0.76). Beetle populations with known resistance to the neonicotinoid imidacloprid also exhibited the highest LC50 levels to metaflumizone suggesting at least the possibility of cross-resistance. Additional experiments using a potato leaf-dip bioassay as well as field efficacy evaluations confirmed the high level of toxicity of metaflumizone to L. decemlineata and demonstrated a potential benefit of tank mixing a low rate of the pyrethroid esfenvalerate with metaflumizone at one-tenth the recommended field rate. These research findings confirm that metaflumizone is highly active against L. decemlineata larvae and adults and could provide an effective alternative insecticide for potato pest management.     

Bioassay determination of the distribution of imidacloprid in potato plants: implications to resistance development

Soil-applied imidacloprid exhibits exceptional efficacy as a systemic insecticide against the Colorado potato beetle, Leptinotarsa decemlineata (Say). An uneven distribution of the chemical within potato plants could result in differential concentrations, which may allow for discrimination between genotypes of varying susceptibility. In this study, susceptible and tolerant larvae were fed leaves from the lower, middle, and upper canopy of treated and untreated plants to characterize within-plant distribution of imidacloprid at 4, 6, 8, 10, 12, and 14 wk after planting. Significant differences in larval mortality and development indicated that the concentration of imidacloprid was unevenly distributed in the potato foliage during 6-14 wk after planting. The concentration of imidacloprid was lowest in the younger tissues of the upper leaves and highest in the older, lower leaves. At 6 wk, a time when the postdiapause beetles are colonizing potato fields, the lower concentration in upper leaves was toxic to susceptible larvae but did not kill a substantial portion of the tolerant larvae. Results suggest that higher concentrations of imidacloprid in the lower canopy leaves may act as a toxic barrier to colonizing susceptible beetles but may allow more tolerant individuals to reach the upper canopy with lower concentrations. Possible scenarios of how different concentrations of the systemic insecticide could influence the rate of resistance development are discussed.     

Overwintering biology ofMyiopharus aberrans andMyiopharus doryphorae (Dipt.: Tachinidae) larval parasitoids of the colorado potato beetle (Col.: Chrysomelidae)

The overwintering biology ofMyiopharus aberrans (Townsend) andMyiopharus doryphorae (Riley), tachinid parasitoids of the Colorado potato beetle, was investigated. Sixty seven overwintered Colorado potato beetle adults were dug from the soil before beetle emergence in the spring of 1990 from a field that had been planted to potatoes the previous year. Five adultM. aberrans and two adultM. doryphorae flies were reared from seven beetles. This is the first record of howM. aberrans andM. doryphorae flies overwinter.      

Toxicity of six commercially formulated insecticides and biopesticides to third instar larvae of mushroom sciarid, Lycoriella ingenua Dufour (Diptera: Sciaridae), in New South Wales, Australia

Abstract  Lycoriella ingenua is one of the major pests of cultivated mushrooms, Agaricus bisporus (Lange) Imbach . Insecticide resistance among mushroom sciarid populations has been reported from other countries, and there is a need to determine the toxicity of currently approved and potential pesticides to sustain control of mushroom sciarid populations in Australia. The present study investigated the toxicity of six commercial formulations of insecticides or biopesticides against third instar larvae of L .  ingenua using laboratory bioassays. Insecticide treatments were incorporated into the growing medium for sciarid larvae and the concentration of the pesticide, which killed 90% of the test population (LD₉₀) determined the efficacy of selected insecticides. Triflumuron was the most effective insecticide against L. ingenua with an LD₉₀ of 53.12 mg active ingredient (a.i.)/m² followed by cyromazine (LD₉₀, 179.68 mg a.i./m²) and diazinon (LD₉₀, 261.72 mg a.i./m²). Abamectin and Bacillus thuringiensis ssp. israelensis were ineffective against L. ingenua . Steinernema feltiae , an entomopathogenic nematode, reduced the number of third instar larvae of L. ingenua only when applied at a higher rate (LD₉₀, 732 422 nematodes/m²) than was recommended on the label.     

Toxicity of four systemic neonicotinoids to adults of Anoplophora glabripennis (Coleoptera: Cerambycidae)

As part of the ongoing evaluation of different systemic insecticides for prophylactic treatment of trees, responses of the beetle Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae) to different doses of four systemic neonicotinyl insecticides were studied. Adult beetles were provided with twigs or leaves of trees treated with different concentrations of imidacloprid to evaluate the toxicity of the insecticide through ingestion or contact or through both. Adult beetles also were provided with twigs of host plant treated with clothianidin, dinotefuran, and thiamethoxam to establish dose response of the beetle to these insecticides. Levels of individual insecticides in twigs and leaves were determined by using the "parent" method with high-performance liquid chromatography, and these levels were compared with the applied concentrations to determine their relationship. The LC50 values for detected level of each insecticide in twigs was 5.1 ppm at 24 h, 2.9 at 48 h, and 1.9 ppm at 72 h for imidacloprid; 1.1 ppm at 72 h for clothianidin; 2.2 ppm at 72 h for dinotefuran; and 1.0 ppm at 72 h for thiamethoxam. Our results indicate that mortality of adult beetles resulted not only from the ingestion and contact toxicity but also possibly from the antifeedant effect of imidacloprid.     

Heterogeneity of twoBeauveria bassiana strains revealed by biochemical tests,protein profiles and bio-assays ofLeptinotarsa decemlineata (Col.: Chrysomelidae) andColeomegilla macultata lengi (Col.: Coccinellidae) larvae

Biochemical profiles on API Rapid CH^* strips and protein profiles on polyacrylamide gels in the presence of sodium dodecyl sulfate were used to distinguish two strains of the entomopathogenic fungusBeauveria bassiana (Balsamo) Vuillemin, ARSEF 2991 and ATCC 44860. Next, the toxicity of these two strains was determined at concentrations of 10², 10⁴, 10⁶ and 10⁸ blastospores/ml on larvae of the Colorado potato beetleLeptinotarsa decemlineata Say (Coleoptera: Chrysomelidae) and of its predator, the spotted ladybird beetle,Coleomegilla maculata lengi Timberlake (Coleoptera: Coccinellidae). Both strains were highly toxic toL. decemlineata larvae. However, the two strains exhibited different levels of toxicity forC. maculata larvae: ARSEF 2991 was toxic, whereas ATCC 44860 caused little coccinellid larval mortality.

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Résumé Les profils biochimiques sur galeries API Rapid CH^* et les profils protéiques sur gels de polyacrylamide ont été utilisés pour distinguer deux souches du champignon entomopathogèneBeauveria bassiana (Balsamo) Vuillemin. La toxicité de ces deux souches a été déterminée à des concentrations de 10², 10⁴, 10⁶ et 10⁸ blastospores/ml sur des larves du doryphore,Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae) et de la coccinelle maculéeColeomegilla maculata lengi Timberlake (Coleoptera: Coccinellidae). Les deux souches deB. bassiana se sont avérées actives à l'égard des larves deL. decemlineata. Toutefois la souche ARSEF 2991 s'est avérée pathogène pour les larves deC. maculata, alors que la souche ATCC 44860 a provoqué une faible mortalité des larves.