The internal microbial flora of the elm bark beetle, Scolytus scolytus, at all stages of its development

Adult Scolytus scolytus emerged from the pupal stage with sterile guts although microorganisms were present within the pupal body. Gut sterility was maintained in a large proportion of newly formed adults while they were boring out of an elm log. The adult gut microbial flora is mainly acquired during subsequent feeding. Differences were found between the microbial gut flora of both larvae and adult S. scolytus, and of both male and female beetles.     

Laboratory studies on the fungus Verticillium lecanii, a larval pathogen of the large elm bark beetle (Scolytus scolytus)

From 1972 to 1974, estimates of the natural larval mortality (> second instar) of elm bark beetles caused by pathogenic organisms were always below 7'5 % of the beetle population. The pathogenic fungus Verticillium lecanii was frequently isolated from field-collected dead larvae, and in the laboratory all larvae were killed in 5 days when exposed to spore concentrations of 4·5 × 10⁶ spores/ml. V. lecanii begins to lose its pathogenicity after prolonged culture on artificial media. The time taken for V. lecanii to kill Scolytus scolytus larvae when exposed to a logarithmic series of spore dilutions from 9·1 × 10⁷/ml to 9·1 × 10³/ml increased with decreasing amounts of inoculum. Even at spore concentrations as low as 9·1 × 10³/ml the mortality of treated larvae was greater than that of untreated individuals. At 100% r.h. all treated larvae were killed over a temperature range of 5–30 °C; those maintained at 25 °C were killed most rapidly and those kept at 5 °C the slowest.     

Comparative laboratory studies on three fungal pathogens of the elm bark beetle Scolytus scolytus: Pathogenicity of Beauveria bassiana,metarhizium anisopliae, and Paecilomyces farinosus to larvae and adults of S. scolytus

The entomogenous fungi Beauveria bassiana (nine isolates), Metarhizium anisopliae (seven isolates), and Paecilomyces farinosus (four isolates) were tested as pathogens of larvae of the elm bark beetle, Scolytus scolytus. Single isolates of B. bassiana and M. anisopliae were also tested against adult beetles. Of the 21 isolates tested as conidial suspensions against larvae, all proved pathogenic. The three most and least virulent isolates were, respectively, isolates of B. bassiana and M. anisopliae. The other isolates fell between these two extremes, with the four P. farinosus isolates all moderately virulent. Spore retention on larvae following inoculation was estimated by washing conidia off the larvae. From the results it was possible to relate larval mortality to the approximate spore dose causing infection at different spore concentrations. Thus, application of spores of the three pathogens at a concentration of 10³ spores/ml resulted in limited mortality. At this concentration, an average of only a single spore was recovered from the inoculated larva. Adult bark beetles also proved susceptible to infection by isolates of B. bassiana and M. anisopliae. They were exposed to discs of elm bark dipped in a conidial suspension. It was estimated that a dose of less than 100 spores could cause infection of beetles following feeding on the elm bark discs.     

More on the life cycle of Malamoeba scolyti (Amoebidae: Sarcomastigophora) parasitizing: the bark beetle Dryocoetes autographus (Scolytidae,Coleoptera)

Some new developmental features previously unreported in, the life- cycle of an amoeba Malamoeba scolyti, which cause cysts in Malpighian tubules of the bark beetle Dryocoetes autographus, are described by means of light, phase-contrast, Nomarski interference, and electron microscopy. Two developmental phases are presumed to take place in the life cycle of M. scolyti: a phase with large multinucleate amoebae (trophozoites) developing in the gut and caecum of the host, and a phase (parasitic phase) causing cysts in the Malpighian tubules. Some data on the host-parasite relationship and rate of infection are also discussed.     

Kinetics of larval gregarious behavior in the bark beetleDendroctonus micans (Coleoptera: Scolytidae)

This paper provides a simple mathematical model of the aggregation behavior of the gregarious intracortical-feeding larvae of the bark beetle, Dendroctonus micans. The model's assumptions are that each individual produces pheromones at a constant rate and reacts positively to a pheromone gradient. These hypotheses have been tested by comparing experiments and simulations, both of which showed that (1) homogeneously distributed individuals aggregate rapidly at the center of the experimental or theoretical arena and (2) eccentric, preformed groups succeed in attracting dispersed individuals, provided that the initial size of these groups is sufficiently high. There is good agreement between most of the experimental and theoretical results, providing a link among chemical communication, density of larvae, and random events occurring during the development of a spatial structure.     

Helicosporidium infection of the great European spruce bark beetle, Dendroctonus micans (Coleoptera: Scolytidae)

An infection of the great European spruce bark beetle Dendroctonus micans (from Turkey) by the parasitic green alga Helicosporidium is described. This is the first time that Helicosporidium has been found to infect a bark beetle (Scolytidae) and the first time that a naturally infected beetle has been reported for the Eurasian continent. The typical cysts of Helicosporidium contain three ovoid cells and one helical, filamentous cell. The morphological characteristics are revealed by light and electron microscopy. Distinct electron-dense inclusions with a peculiar ultrastructure may represent pyrenoids. Since D. micans is an important pest, the discovery of a natural pathogen may offer a chance for biological control.     

Effects of the bark beetle nematode, Contortylenchus reversus, on gallery construction,fecundity, and egg viability of the Douglas Fir beetle, Dendroctonus pseudotsugae (Coleoptera: Scolytidae)

Healthy and Contortylenchus reversus infected Douglas Fir beetles were colonized in logs in the laboratory. At 7, 14, and 23 days after introduction of the beetles into the logs, the bark was removed from some of the logs and the length and shape of the primary gallery, the number of eggs laid, and the egg viability were determined for each pair of parent beetles. Parasitism caused a 25, 28, and 27% reduction in the length of the primary egg gallery built by the infected female at 7, 14, and 23 days, respectively, after the start of colonization. In addition, the number of eggs laid by the infected females by 7, 14, and 23 days after the start of colonization were 50, 33, and 45%, respectively, lower than that of healthy female beetles. Despite the differences in gallery length between control and infected females, linear regression analysis showed that the rates of gallery construction were not significantly different. However, infected female beetles showed significantly lower rates of egg-laying than did healthy female beetles. Nematode parasitism did not affect gallery shape and egg viability of female Douglas Fir beetles. Mating of healthy female beetles with infected males affected neither the gallery length nor the fecundity of the uninfected females.     

Screening of entomopathogenic fungi against the European spruce bark beetle,Dendroctonus micans (Coleoptera: Scolytidae)

The European spruce bark beetle, Dendroctonus micans Kugelann (Coleoptera: Scolytidae), is one of the most serious destructive pests of oriental spruce (Picea orientalis L.). In order to find an effective biocontrol agent against this pest, we determined the effectiveness of entomopathogenic fungi against D. micans. Virulence of nine highly pathogenic strains including Beauveria bassiana (2), Beauveria cf. bassiana (2), Metarhizium anisopliae (2), Metarhizium sp. (1), Isaria fumosorosea (1) and Evlachovaea sp. (1) was evaluated on D. micans larvae and adults under laboratory conditions. Mortality values for both larvae and adults ranged from 83 to 100% and from 23 to 100%, respectively. Larvae were more susceptible to fungi than adults (P<0.05). Based on screening tests, B. cf. bassiana isolate KTU-53 was found the most effective isolate. LC₅₀ values were calculated as 1.77×10⁴ and 2.65×10⁴ conidia mL^?1 for isolate KTU-53 against larvae and adults, respectively. Consequently, B. cf. bassiana isolate KTU-53 appears to be the most promising microbial control agent for biocontrol of D. micans.     

Effect of different semiochemicals blends on spruce bark beetle,Ips typographus (Coleoptera: Scolytidae)

The spruce bark beetle, Ips typographus, is a recent new introduction to the Qilian Mountains of China. An outbreak of these beetles has infested over 0.03 million hectares of spruce forests in this area. Although primary attraction to volatiles has been clearly demonstrated for I. typographus, the existence and role of attraction to insect‐produced pheromones have been widely debated. Currently, commercial lures for I. typographus include only the volatiles ipsdienol, cis‐verbenol, trans‐verbenol, 2‐methyl‐3‐buten‐2‐ol and 2‐phenylethanol in Europe. Several potential pheromone candidates have been identified for I. typographus. Our GC–MS and GC–FID analyses volatiles from hindgut extracts of I. typographus in different attack phases demonstrated that the 2‐methyl‐3‐buten‐2‐ol, ipsdienol, cis‐verbenol and trans‐verbenol as major hindgut components, and ipsenol, 2‐phenylethanol, trans‐ myrtenol and verbenone as minor components. We tested various combinations of semiochemical candidates, to determine an optimal blend. Our results suggest that addition of 2‐methyl‐3‐buten‐2‐ol to either ipsenol alone, or to blends of ipsenol and other semiochemical candidates, significantly enhanced attraction of I. typographus. Therefore, a simple lure consisting of ipsenol and 2‐methyl‐3‐buten‐2‐ol would be an optimal blend of I. typographus in the Qilian Mountains, China. We conclude that this optimal semiochemical blend may provide an effective biological pest control method for use in forest ecosystem against I. typographus.     

Comparative efficacy of different pesticides against mango bark beetle Hypocryphalus mangiferae Stebbing (Coleoptera: Scolytidae)

Hypocryphalus mangiferae Stebbing is one of the most destructive insect pests of mango trees and is found to be associated with the transmission of causal organisms of mango sudden death disease. The present study was carried out to evaluate the toxicity of deltamethrin, bifenthrin, chlorpyrifos, emamectin benzoate, imidacloprid and spinosad in laboratory and field trials. Bioassay results showed that the toxicity of chlorpyrifos was significantly higher than deltamethrin but similar to bifenthrin. Deltamethrin and bifenthrin toxicity, however, increased significantly (P < 0.01) from day 1 to day 3. Spinosad was the least toxic compound while emamectin was the most toxic among new chemical insecticides tested, but its toxicity increased significantly from day 1 to day 5. Comparison of the efficacies of the insecticides using lethal times to produce 50% mortality (LT₅₀) and 90% mortality (LT₉₀) showed that the relative potencies of chlorpyrifos, emamectin, imidacloprid and spinosad were greater than bifenthrin and deltamethrin. The results of field trials showed the highest number of beetles emerged from the control twigs while significantly fewer beetles emerged from the twigs treated with bifenthrin (P < 0.05), which accounted for 12% for bifenthrin compared to that of the control. The present study demonstrated increased toxicities of systemic insecticides and chlorpyrifos compared to toxicities of deltamethrin and bifenthrin, suggesting these insecticides could be an alternative tool in a comprehensive H. mangiferae management program to eradicate the beetles from mango orchards.     

Degradation of the isoflavone biochanin a by isolates of Nectria haematococca (Fusarium solani)

Twelve isolates of Nectria haematococca, mating population VI (Fusarium solani) previously characterized for their virulence on pea plants and their ability to degrade the phytoalexin pisatin were assayed for the catabolism of the isoflavone biochanin A (5,7-dihydroxy-4′-methoxyisoflavone). Eleven isolates catabolized the isoflavone along the pathway: biochanin A → dihydrobiochanin A → 3-(p-methoxyphenyl)-6-hydroxy-γ-pyrone → p-methoxyphenylacetic acid → p-hydroxyphenylacetic acid → 3,4-dihydroxyphenylacetic acid.     

The first study on the bacterial flora of the european spruce bark beetle, Dendroctonus micans (Coleoptera: Scolytidae)

The European spruce bark beetle, Dendroctonus micans Kugelann (Coleoptera, Scolytidae), is one of the most serious pests of oriental spruce (Picea orientalis L.) in Turkey. In this study, we investigated bacterial flora of D. micans collected from different populations of the forests of Eastern Black Sea Region of Turkey from 2002 to 2004. Seven different bacteria were isolated from healthy, diseased and dead specimens based on the color of colony and morphology. According to morphological, physiological and biochemical properties, metobolic enyzme profile by BIOLOG microtiter plate system, and total cellular fatty acid profile by Microbial Identification System (MIS), isolates were identified as Micrococcus luteus, Bacillus thuringiensis subsp. morrisoni, Serratia grimesii, Enterobacter cloaceae, Enterobacter intermedius, Streptococcus sp. and Pseudomonas putida. This is the first study on the bacterial flora of D. micans.     

Isolation and virulence of entomopathogenic fungi against the great spruce bark beetle,Dendroctonus micans (Kugelann) (Coleoptera: Scolytidae)

Twelve fungal strains including Lecanicillium muscarium (Petch.) Zare and Gams, Isaria farinosa (Holmsk.) Fr., Fusarium sp., Beauveria bassiana Sensu Lato and Beauveria sp. were isolated from larvae and adults of D. micans. In addition, virulence of these isolates against this pest was determined. Conidia suspensions of 1×10⁶ conidia mL^–1 were applied to larvae and adults. The highest mortality and mycosis for larvae were obtained from isolate ARSEF 9271 (Beauveria bassiana) with 90% mortality and mycosis within 10 days. ARSEF 9271 also produced 93% mortality and mycosis in adults. On the other hand, the highest mortality and mycosis for adults were obtained with isolate ARSEF 9272 (Beauveria sp.), with 100% mortality and 80% mycosis within 10 days. These results indicate that isolates ARSEF 9271 and ARSEF 9272 seem to be the most promising potential fungal biocontrol agents against D. micans.     

Flight initiation and survival in the bark beetle Ips typographus (Coleoptera: Scolytidae) during the spring dispersal

Temperatures in the forest litter of Norway spruce Picea abies were recorded throughout the day to obtain environmental parameters that could be used to design realistic flight-activity experiments in the laboratory. Flight activity and survival were monitored electronically in plastic chambers where the conditions were controlled by an environmental chamber. Flight attempts of the bark beetle Ips typographus were initially lower in chambers with forest duff but were prolonged compared with those of beetles in chambers with a metal screen substrate. Small bark slabs and spruce twigs in the duff were utilized as food and extended the period of flight and survival. A thermal gradient in duff from 25° at the surface down to 13.8°C at a depth of 4 cm also slightly increased the survival of beetles compared with a constant 25°. A daily ambient temperature cycle as well as the duff thermal gradient increased the survival from about 3 d to more than 8 d. The latter length in the laboratory agreed with survival rates of caged beetles in a clearcut forest area, while beetles caged in the forest survival for more than 14 d. Temperatures were monitored at the duff surface of the caged beetles and compared with the catches of beetles that were attracted to a pheromone trap and collected with an electronic fraction collector. Information on flight and survival during the dispersal period is necessary to the design of ecologically sound management programs for control of bark beetles.     

Gallery construction and oviposition of the bark beetle Ips typographus (Coleoptera: Scolytidae) at different breeding densities

Abstract. 1. The bole of a Norway spruce, Picea abies (L.) Karst., was cut in 25 cm sections and infested in the laboratory with 2 or 8 females/dm² of Ips typographus (L.) and an equal number of males.
2. One log of each beetle density was dissected every second day, beginning 2 days after female introduction.
3. The oviposition rate was highest at the beginning of the breeding period for both densities and then declined, especially at the high density. Residence time is a useful approximation for the number of laid eggs in the beginning of the oviposition period or when the density is low.
4. Final gallery length and egg number per gallery were on average significantly smaller at density 8 than at density 2.
5. Linear relationships between total gallery length and number of egg niches were found to be similar for the two densities.
6. An estimate of the egg to adult mortality at different densities was found by using data from a field study that related gallery length to breeding density in attacked living trees, together with published relationships between density and egg niches or emergence of new adults.
7. It was concluded that both decreased oviposition rate and earlier re-emergence at higher densities contribute to a declining oviposition with increasing density. The final offspring production is futher reduced by lower survival due to larval competition as density increases.     

Nutrient-mediated germination of Beauveria bassiana conidia on the integument of the bark beetle Dendroctonus ponderosae (Coleoptera:Scolytidae)

Field-collected adult mountain pine beetles, Dendroctonus ponderosae, were inoculated with the white muscardine fungus, Beauveria bassiana, and thoroughly examined externally with a scanning electron microscope. Germinating conidia were found at a very low incidence, and only on antennal clubs. However, mortality of inoculated controls was high, and B. bassiana was confirmed as the causative agent. Germination on the cuticle was greatly increased by sonication of adult beetles. The hypothesis that hemolymph released through sonication-damaged membranes provided a nutrient stimulus that enhanced conidial germination was tested by placing hemolymph or yeast-malt extract broth on the cuticle of otherwise nontreated beetles, which were then inoculated with conidia. Significant germination occurred on the cuticle of these beetles. Therefore, a limiting factor for conidial germination on the sclerotized cuticle of D. ponderosae is probably a lack of sufficient nutrients.     

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.     

Feeding behaviour of the black pine beetle,Hylastes ater (Coleoptera: Scolytidae)

1 The feeding behaviour of Hylastes ater was investigated at 15 °C and 20 °C on a range of tree species. The role of an aggregation pheromone and the part played by olfactory cues in food selection was also investigated. 2 A distinct hierarchical preference at both 15 °C and 20 °C such that Pinus radiata > Pinus sylvestris > > Picea abies > > Rubus fruticosus=Betula pendula was shown. 3 Feeding on P. sylvestris led to the highest weight gain, closely followed by P. radiata and P. abies. Slightly elevated feeding was observed at 20 °C compared with 15 °C. 4 Reduced feeding in the presence of B. pendula suggests the possibility of an antifeedant effect and predamaged wood was found to be less nutritious, possibly due to greater plant defences. 5 The role of an aggregation pheromone was not supported. Olfactory cues enabled the detection of woody hosts, although differentiation between host was not observed. 6 These results are likely to be important in helping to design an effective integrated control approach against this pest.     

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.