Some effects of freezing temperatures on overwintering larvae of the large elm bark beetle (Scolytus scolytus)

Overwintering final instar larvae of Scolytus scolytus were tested for coldhardiness by exposure to a range of sub-zero temperatures (–7°Cto –31 °C) in a frost-gradient apparatus for 7 days. The Lt₅₀ for larvae removed from the bark (– 20.5 °C) was significantly different (P < 0.01) from the Lt50 (–18.3 °C) for larvae insulated by the bark (thickness of 7 mm ± 2 mm). Larvae with food in their digestive tract were more susceptible to freezing than the overwintering final instars which had voided their stomach contents. Duration and intensity of cold did not cause any adverse long-term effects. Most of the larvae which survived the sub-zero treatments were able to pupate or reach the adult stage. The mean supercooling point of the overwintering larvae (–30.85 °C) confirmed their cold-hardiness.     

The susceptibility of the elm bark beetle Scolytus scolytus to the DD-136 strain of Neoaplectana sp.

Adults and larve of Scolytus scolytus were highly susceptible to the DD-136 strain of Neoaplectana sp. in the laboratory.     

The DD-136 strain of Neoaplectana sp. as a potential biological control agent for the European elm bark beetle,Scolytus scolytus

When logs infected with Scolytus scolytus were sprayed with the DD-136 strain of Neoaplectana sp. in solutions of either distilled water, glycerin, or P.B.I. wetting agent, the nematodes were efficiently dispersed and entered the logs through the insect entrance holes in the bark.Logs treated with Neoaplectana sp. were found to contain a significantly higher proportion of dead, nematode-infected insects. No difference was found between the various methods of nematode application. The results indicate that Neoaplectana sp. may have a possible role in the control of S. scolytus in elm logs.     

Fusarium solani, a weak pathogen of the larval stages of the large elm bark beetle Scolytus scolytus (Coleoptera: Scolytidae)

Fusarium solani, a common colonist of beetle-infested elm bark, has been isolated from dead larvae and adults of the large elm bark beetle, Scolytus scolytus. Two infectivity tests indicated that this fungus is probably a weak pathogen of the larval stages of the beetle. Total mortality was not achieved in either test, but the mortality associated with fungal inoculation was occasionally significantly greater than the mortality in the uninoculated controls in both experiments (P<0.05, experiment A; P<0.025, experiment B). Disease symptoms suggest that the larvae attacked by F. solani may have died from bacterial infection.     

Behavioral responses of the smaller European elm bark beetle, Scolytus multistriatus, to extracts of non-host tree tissues

Crude benzene extracts of 6 non-host trees alone did not stimulate feeding responses by adults of Scolytus multistriatus (Marsh.). When combined equally with a stimulatory extract from Ulmus americana L., all of the crude non-host extracts reduced the amount of feeding which otherwise occurred. At least two of the non-hosts (Populus deltoides Bartr. and Aesculus octandra Marsh) contained positive short-range orientational stimuli, but their induction of a positive response by the insect was prevented in most treatments by associated deterrent chemicals. Thus, stimuli (deterrents and repellents) played the dominant role in eliciting the rejection of crude non-host extracts by the beetles.

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Zusammenfassung Benzol-Rohextrakte von 6 Nicht-Wirtsbäumen riefen allein bei erwachsenen Scolytus multistriatus (Marsh.) keine Fraßreaktionen hervor. Wurden sie zu gleichen Teilen mit einem stimulierenden Extrakt von Ulmus americanus L. kombiniert, so setzten alle Rohextrakte aus Nicht-Wirten die Fraßmenge herab, die sonst auftritt. Wenigstens zwei der Nicht-Wirte (Populus deltoides Bartr. und Aesculus octandra Marsh.) enthielten auf kurze Entfernung wirkende positive Reize, aber ihr Einfluß auf eine positive Reaktion des Insekts wurde bei den meisten Behandlungen durch Verbindung mit abweisenden (deterrent) Stoffen verhindert. So spielen bei der Auslösung der Ablehnung von Rohextrakten der Nicht-Wirte durch die Käfer Deterrent- und Repellent-Reize die Hauptrolle.

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Approved for publication by the Director of the Wisconsin Agricultural Experiment Station. This research was supported in part by a research grant (GB-6580) from the National Science Foundation; and from funds provided by the State of Wisconsin Department of Natural Resources.     

Control of aphids in glasshouses with the fungus, Verticillium lecanii

In small glasshouses, aqueous sprays of spores of the entomopathogenic fungus, Verticillium lecanii , eliminated small populations of the aphid Brachycaudus helichrysi in the vegetative tips of chrysanthemum plants but not when they were in an exposed position on mature flower buds. Control of the chrysanthemum aphid, Macrosiphoniella sanborni , was however variable and usually commercially unsatisfactory. In contrast, in both small and large glasshouses, sparse populations of the major aphid pest Myzus persicae were successfully and consistently controlled, sometimes spectacularly. The inherent susceptibilities of these aphid species, measured by laboratory bioassay, did not account for the differences in control observed in the glasshouse. It is thought that a combination of species-characteristic feeding site preferences on the exposed parts of plants, where microclimate humidity was probably low, and relative immobility of M. sanborni and B. helichrysi explain why these aphids were less well-controlled than My. persicae . However, it should be possible to control satisfactorily all aphid species in large commercial glasshouses where humidity might be higher than in the small experimental glasshouses. A single spray was sufficient to introduce infection that controlled aphids for the duration of the crop.     

Volatiles released from oak, a host tree for the bark beetle Scolytus intricatus

Volatile compounds emitted in different phases of oak (Quercus robur) development (bark, unopened buds, young developing leaves, and blossoms) were analyzed with the aim of finding possible host-plant attractants for the European oak bark beetle, Scolytus intricatus. Complex mixtures of aliphatic, aromatic, and terpenoid compounds were identified in the samples. (E)-2-Hexenal and hexanal dominated in samples of bark. In buds, (Z)-3-hexenyl acetate formed a substantial part of the mixture. In both leaves and blossoms (E,E)--farnesene was the main component.

Volatiles released from oak twigs and branches during both the maturation feeding and construction of maternal galleries by Scolytus intricatus were also analyzed. Most compounds found in the samples from females’ and males’ maturation feeding were identical. High contents of anisole, (E)-β-ocimene, -copaene, one unidentified sesquiterpenic hydrocarbon C₁₅H₂₄ and β-caryophyllene were found in both samples of twigs attacked by beetles. During the construction of maternal galleries by bark beetles in oak logs, monoterpene hydrocarbons such as p-cymene, (E)-β-ocimene, and γ-terpinene, and sesquiterpenes -copaene and β-caryophyllene were released in large quantities. No new compound appeared when males were added to the log with feeding females.     

Genetic,morphological, and virulence characterization of the entomopathogenic fungus Verticillium lecanii

In order to clarify relationships among genetic diversity, virulence, and other characteristics of conidia, 46 isolates of Verticillium lecanii from various hosts and geographical locations were examined. The internal transcribed spacer (ITS) and intergenic spacer (IGS) regions of ribosomal DNA (rDNA), mitochondrial small subunit rDNA (mt-SrDNA) and beta-tubulin were analyzed by PCR-RFLP. PCR-single stranded conformational polymorphism (SSCP) was performed on regions of the mitochondrial large subunit rDNA, mt-SrDNA, beta-tubulin and histone 4. There were no relationships among the results of RFLP, SSCP, isolation source, and location. However, amplified product size of IGS did have relationships with conidia size and sporulation. Six isolates with 4.0-kb IGS products had large conidia dimensions, and yielded low numbers of conidia compared with other isolates. Three out of the six isolates were high virulence (over 90%) against green peach aphids. Furthermore, double-stranded RNA (dsRNA) was detected in 22 out of 35 V. lecanii isolates and related with the amplicon sizes of IGS, though not with virulence or isolation location. Isolates containing dsRNA were divided into six distinct types based on banding pattern. These data demonstrate the level of genetic diversity of V. lecanii, and suggest relations among the genetic properties and conidial morphology.     

Epizootic potential for aphids of different isolates of the fungus,Verticillium lecanii

Large (6.7–8.4 μm in length) and small (3.8–6.7 μm in length) spored strains ofVerticillium lecanii (Zimm) Viégas killed similar numbers of adults of the aphids treated with aqueous spore suspensions. However, when these strains were compared by determining the proportions of progeny aphids acquiring infection from treated adults, i.e. a measure of epizootic potential, large differences emerged; only large-spored strains exhibited as strong an epizootic potential as the standard strain, 1–72, with mean progeny mortality ratios (test/standard) close to 1.0. Further experiments indicated that the speed of germination, fastest for large-spored strains, accounted for the greater epizootic potential of such strains. In addition, the mode of sporulation of large-spored strains on aphid cadavers probably also contributed to the more efficient spread of such strains. A general conclusion was reached that measurement of the effectiveness of a pathogen against a host insect must be based on many factors, in addition to virulence against a single developmental stage.     

The role of beetle and host volatiles in host colonization in the European oak bark beetle, Scolytus intricatus (Ratzeburg) (Col., Scolytidae)

Abstract:  To understand the role of chemical signals involved in Scolytus intricatus (Ratzeburg) (Col., Scolytidae) host colonization, the attractiveness of intact and beetle colonized host material was investigated in the field and in the laboratory. In the field, chemical signals operating at long range were investigated by means of sticky traps. In the laboratory, close-range chemical interactions were investigated with an arena olfactometer. Field experiments showed no differences in the attractiveness between infested and non-infested host material. On the contrary, laboratory experiments revealed higher attractiveness of infested host and beetle-produced frass in comparison with intact host and mechanically produced sawdust respectively. Laboratory data also disclosed the attractiveness of beetle extracts of both sexes. Our data show that: (1) host kairomones play an important role during host colonization, and that (2) S. intricatus does not use a sex-specific secondary attractant system. Differences between results of field and laboratory trials are discussed.     

Pathogenicity of the fungus, Verticillium lecanii, to the green peach aphid, Myzus persicae (Hom.: Aphididae)

Pathogenicity of the hyphomycete, Verticillium lecanii (DAOM 198499), was investigated to aphid, Myzus persicae under laboratory conditions. Analysis of lethal effect of six various concentrations 0, 10(4), 10(5), 10(6), 10(7) and 10(8) conidia/ml of V. lecanii against third nymphal stage of M. persicae, indicated significant mortality on aphids. Mean comparisons showed that there was significant difference between treatments. Three days after treatment, aphid mortality observed and after 12 days minimal mortality was 17.77 in control and maximum was 100 percent related to 10(7) and 10(8) conidia/ml. LC50 and LT50 values were estimated by probit analysis and life test. LC50 value for aphid mortality was 1.4 x 10(4) conidia/ml, LT50 values for concentrations 10(4), 10(6), 10(6), 10(7) and 10(8) conidla/ml was 10, 10, 9, 8 and 6 days, respectively. In this experiment, net reproduction rate of aphid's (R0) decreased significantly when concentration increased. These observations showed that V. lecanii (DAOM 198499) can be an active biological agent against aphids.     

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.     

Germination of the entomopathogenic fungus Verticillium lecanii on scales of the glasshouse whitefly Trialeurodes vaporariorum

In a series of laboratory experiments, tobacco leaf discs infested with scales of the glasshouse whitefly Trialeurodes vaporariorum, were immersed in suspensions of conidia of three strains of the entomopathogenic fungus Verticillium lecanii. The germination of conidia on the insect was recorded and compared with the germination of conidia applied to Czapek—Dox Complete Medium. Laboratory bioassays were performed to record the mortality of whitefly scales exposed to conidia of all three fungal strains. The rate of germination of conidia on the whitefly scales was much reduced compared to that oh complete medium. Differences were recorded in the rate of germination of the three strains on complete medium, but not on the whitefly scales. There was no correlation between the pathogenicity of the strains to whitefly scales in laboratory bioassays and the rate of germination on the host.     

Significance of parasitoids in the reduction of oak bark beetle Scolytus intricatus Ratzeburg (Col., Scolytidae) in Serbia

Abstract:  As the vector of vascular fungi of the genus Ophiostoma the oak bark beetle, Scolytus intricatus , is one of the significant links in the chain of agents of oak forest decline in Serbia. It is known that the adults of this bark beetle, which develop under the bark of the trees infected by fungi of the genus Ophiostoma , transport the spores of these fungi. During the maturation feeding, the spores are transmitted to healthy tree crowns, where they germinate and cause infection. In period 1992–96 at 27 localities in Serbia, the significance of parasitoids in the reduction of this bark beetle was examined, as the intensity of spore transmission depends upon the number of oak bark beetle adults in nature. In the research of parasitoids of S. intricatus 20 species were identified in five families of Hymenoptera: Braconidae (six species), Eurytomidae (one), Pteromalidae (10), Eupelmidae (one) and Eulophidae (two). Among these parasitoids the greatest influence on the abundance of oak bark beetle was the species Ecphylus silesiacus . It was identified in 90.91% of study samples, its domination was 39.15% and the percentage of oak bark beetle parasitism was 5.66%. In addition, the species Entendon ergias , Dendrosoter protuberans and Cheiropachus quadrum were significant in reduction of S. intricatus . The average percentage of parasitism of oak bark beetle by E. ergias was 2.74%, by D. protuberans it was 2.63% and by Ch. quadrum 1.63%. The significance of other parasitoids in the reduction of oak bark beetles was low. In the study period the total parasitism of oak bark beetle varied between a minimum of 8.64% in 1992 and a maximum of 19.64% in 1996. The average for the whole study period was 14.49%.     

Comparative laboratory studies on three fungal pathogens of the elm bark beetle Scolytus scolytus: Effect of temperature and humidity on infection by Beauveria bassiana,Metarhizium anisopliae, and Paecilomyces farinosus

Larvae of the elm bark beetle, Scolytus scolytus, were inoculated with conidia of the entomogenous fungi Beauveria bassiana (two strains), Metarhizium anisopliae (two strains), and Paecilomyces farinosus (two strains) and incubated over a range of temperatures (2°, 6°, 10°, 15°, and 20°C). One strain each of B. bassiana and P. farinosus caused infection even at 2°C, whereas the two strains of M. anisopliae caused no infection below 10°C. Infection of adult beetles by B. bassiana (one strain) and M. anisopliae (one strain) was tested at 15°, 20°, and 25°C (B. bassiana) and at 15° and 20°C (M. anisopliae). Fungal infection occurred at all three temperatures, but at 25°C beetles tended to succumb to bacterial infection. The effect of relative humidity on infection of larvae by B. bassiana (one strain), M. anisopliae (one strain), and P. farinosus (one strain) was tested at 51, 74, 86, 90, 95, 97.5, and 100% relative humidity. B. bassiana and M. anisopliae caused some infection at all humidities: with P. farinosus there was no infection at the two lowest humidities. Mortality due to infection by these fungi was most rapid at the highest humidities.     

Control of glasshouse whitefly, Trialeurodes vaporariorum, by an 'aphid' strain of the fungus Verticillium lecanii

Heavy infestations of whitefly on glasshouse cucumber plants were controlled below the level of economic crop damage by fortnightly or monthly sprays of Verticillium lecanii spores. The fungus did not spread from glasshouse to glasshouse, or from plant to plant, and often not from diseased whitefly scales bearing fungal spore heads to nearby healthy scales. Some scales survived and the resulting adults laid eggs on new leaves bearing no infected scales, creating another, healthy, generation. This makes regular spraying of new leaves essential. Blastospores were as effective as conidia in controlling scales when sprayed to ‘run off’ at concentrations near 10⁷ spores ml^-1 sprayed on to the undersurfaces of leaves. A fivefold increase in spore concentration at levels near 10⁷ spores ml^-1 usually caused significant improvement in mortality, but increase above this concentration is likely to be unrewarding. Thorough coverage of leaves was found to be vital. Control was impaired by dry conditions and by prolonged air temperatures above 25 °C. The fungicide dimethirimol, used against cucumber mildew, did not impair whitefly control by V. lecanii.     

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

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

The effect of larval aggregation behaviour on larval growth of the spruce bark beetle Dendroctonus micans

1. The great spruce bark beetle Dendroctonus micans is a primary pest of spruce in Europe. It is unusual among Eurasian scolytids in that apparently healthy trees are attacked by solitary adults, but larvae feed en masse , in response to a larval aggregation pheromone.
2. The effect of brood size on larval growth was determined in experiments on detached bark, logs and living trees. A positive relationship was found between brood size and larval growth in experiments started with either eggs or larvae up to fifth instar. The relationship appeared to be independent of the effects of both preformed and induced defences in bark.
3. No evidence was found to support the hypothesis that larvae feeding in groups spend a greater proportion of time feeding. Addition of resin to egg chambers in detached bark pieces resulted in high mortality of first-instar larvae. A possible role for larval aggregation in minimizing the effects of host defences is discussed.