<Emphasis Type="Italic">Bursaphelenchus mucronatus</Emphasis> (Nematoda: Aphelenchoididae) vectored by <Emphasis Type="Italic">Monochamus </Emphasis><Emphasis Type="Italic">urussovi</Emphasis> (Coleoptera: Cerambycidae) in Hokkaido,Japan

Bursaphelenchus mucronatus Mamiya et Enda has been recovered for the first time from adults of the cerambycid beetle, Monochamus urussovi (Fischer), in Hokkaido, Japan. The nematode was also recovered from the inner bark of Picea jezoensis (Siebold et Zuccarini) Carrière and Abies sachalinensis (Fr Schmidt) Masters infested with M. urussovi larvae. PCR–RFLP analysis indicated that B. mucronatus in Hokkaido is the European type.     

Population dynamics of <Emphasis Type="Italic">Monochamus galloprovincialis</Emphasis> Olivier (Coleoptera: Cerambycidae) in two pine species under laboratory conditions

The study of the life history dynamics of the pine sawyer beetle Monochamus galloprovincialis (Olivier) is important because the beetles vector the pinewood nematode. The objectives of this study were to investigate differences in beetle life history parameters between the two common host tree species in Turkey, Pinus sylvestris and Pinus nigra. Relationships between the number of oviposition sites, holding time (time between log cut and oviposition), log moisture content, log volume and area, and adult survivorship were evaluated. No significant differences were found between the two tree species except adult density per m². The holding time did not influence the number of oviposition sites, larval entry holes or adults for P. sylvestris. There was a negative correlation between the holding time and the number of oviposition sites for P. nigra. The number of adults emerged per log was correlated with the number of larval entry holes only for P. sylvestris. Only 12 and 15% of the initial cohort completed development and emerged as adults for P. sylvestris and P. nigra, respectively. Both tree species are equally suitable hosts for M. galloprovincialis development. We conclude that the high within-log mortality of beetles is a combination of resource quality and quantity factors and intraspecific competition and cannibalism during larval development.     

Abundance-dependent transmission of the pinewood nematode, <Emphasis Type="Italic">Bursaphelenchus xylophilus</Emphasis> (Nematoda: Aphelenchoididae), to the Japanese pine sawyer, <Emphasis Type="Italic">Monochamus alternatus</Emphasis> (Coleoptera: Cerambycidae), adult in its pupal chamber

The transmission ratio of the pinewood nematode (PWN), Bursaphelenchus xylophilus, to the emerging adult Japanese pine sawyer (JPS), Monochamus alternatus, in its pupal chamber is a determinant of the number of the nematodes carried by JPS beetles. To investigate the factors affecting the transmission ratio, we counted the number of the nematodes carried by 36 newly emerged JPS beetles and the number remaining in and around their pupal chambers, and then estimated the transmission ratio (the number of nematodes carried by a JPS adult as a percentage of the total number of nematodes aggregating in and around its pupal chamber). The total number of nematodes aggregating in and around a pupal chamber ranged from 0 to 19,041, and the number of nematodes carried by a beetle ranged from 0 to 18,920. The transmission ratio correlated with neither the water content of the wood around the pupal chamber nor the degree of wood discoloration caused by blue-stain fungus. The transmission ratio varied with the abundance of the nematodes aggregating in and around the pupal chamber. In pupal chambers with more than 1,000 nematodes, almost all the nematodes were transmitted to the beetle. However, in pupal chambers with fewer than 1,000 nematodes, the transmission ratio varied greatly, from 0 to 100%. These results suggest that aggregation of many PWNs in the pupal chamber might stimulate transmission of the PWNs to the JPS adult and that this abundance-related transmission might contribute to the large variation in the number of PWNs carried by the JPS beetle.     

Threshold temperatures and degree-day estimates for development of post-dormancy larvae of <Emphasis Type="Italic">Monochamus galloprovincialis</Emphasis> (Coleoptera: Cerambycidae)

Developmental thresholds and thermal requirements for development of post-dormancy larvae of Monochamus galloprovincialis (Olivier) (Cerambycidae; Monochamini) were studied at ten constant temperatures ranging from 7 to 35°C. The relationship between temperature and development duration in days was linear between 15 and 30°C (r ²  = 0.98). The lower threshold for development was determined to be 12.2 ± 0.8°C and an average of 822 degree-days (DD) above that value was required for 50% adult emergence under laboratory conditions. The rate of larval development decreased above 30°C and the lethal upper threshold was between 32 and 35°C. Degree-day rate summation was initiated in the first of March and model predictions were validated with records of field emergence for the years 2001 to 2004. The modified sine wave predicted median emergence with an average error of 3.8 days from emergences in the field and a zero-day difference for two of the years. Model predictions were always within 10% of actual observed emergences. Predictions for early emergences (cumulative percentiles 1 and 10) were less accurate than predictions for median and late emergencies. The results suggest that a simple linear method driven by air temperatures can predict the emergence of M. galloprovincialis with sufficient accuracy to improve the pest management programmes currently implemented on the pine wilt disease affected zone in Portugal.     

First detection of Bursaphelenchus mucronatus (Nematoda: Aphelenchoididae) on Monochamus sutor (Coleoptera: Cerambycidae) in Romania

As a result of the detection of the pinewood nematode Bursaphelenchus xylophilus in Portugal, and its subsequent spread to Spain, intense surveys were conducted to screen for the presence of Bursaphelenchus species in Romania. Herein, we report recent surveys of insects potentially vectoring Bursaphelenchus species collected using trap trees or pheromone‐baited traps placed in the forest. Trap felled spruce trees (Picea abies) and pheromone‐baited traps were installed in six different counties in Romania (Bra?ov, Sibiu, Suceava, Hunedoara, Timi? and Dâmbovi?a). Ten different species of insects distributed among Curculionidae and Cerambycidae were obtained. Nematodes were extracted from insects and observed to validate the presence of Bursaphelenchus specimens. One female identified as Monochamus sutor was the only specimen carrying nematodes in the genus Bursaphelenchus. Nematodes were identified as B. mucronatus based on morphological and molecular features. This is the first detection and report of natural spread of B. mucronatus in Romania. The absence of B. xylophilus was confirmed in the areas of Romania surveyed in this work.     

Control of a new turf pest, <Emphasis Type="Italic">Dorcadion pseudopreissi</Emphasis> (Coleoptera: Cerambycidae), with the entomopathogenic nematode <Emphasis Type="Italic">Heterorhabditis bacteriophora</Emphasis>

The use of biological control in turf has increased to avoid possible negative effects on humans. Entomopathogenic nematodes (EPNs) belonging to the families Steinernematidae and Heterorhabditidae have control potential against many economically important insect pests. In the present study, the efficacy of Heterorhabditis bacteriophora against a new pest on turf, Dorcadion pseudopreissi, was examined in the field. Cages (1 × 1 × 1 m) with female and male D. pseudopreissi were placed on two kinds of turf: Lolium perenne and Festuca arundinacea. After beetles had deposited eggs into the moist soil surface, the cages were removed and H. bacteriophora was applied at 0.5 million infective juveniles/m² to half the plots. Application of H. bacteriophora caused a statistically significant reduction in numbers of beetle larvae in L. perenne plots and a non-significant reduction in F. arundinacea plots. The area damaged by D. pseudopreissi was significantly reduced by nematode application in both turf species. The number of nematodes declined after application, but small numbers could still be detected after 6 months.     

First report of Bursaphelenchus mucronatus kolymensis (Nematoda: Aphelenchoididae) on Monochamus sutor (Coleoptera: Cerambycidae) in Spain

This is the first report of Monochamus sutor as vector of B. mucronatus kolymensis, the European type of B. mucronatus (synonymous with B. kolymensis) in Spain. In 2013, a female of Monochamus sutor was caught in a Pinus sylvestris forest. Nematode species identification was based on morphological characteristics and on molecular and phylogenetic analysis of dauer juveniles and adults.     

Toxicity of synthetic and biological insecticides against adults of the <Emphasis Type="Italic">Eucalyptus</Emphasis> snout-beetle <Emphasis Type="Italic">Gonipterus</Emphasis><Emphasis Type="Italic">scutellatus</Emphasis> Gyllenhal (Coleoptera: Curculionidae)

Toxicity by contact and by ingestion of lufenuron, flufenoxuron, lambda (λ)-cyhalothrin, cypermethrin, thiamethoxam and five entomopathogenic insecticides (three formulations of Beauveria bassiana, a compound containing spores of Metarhizium anisopliae var. acridum, and a mixture of Brevibacillus laterosporus, Bacillus licheniformis and Bacillus chitinosporus) were evaluated on adults of the Eucalyptus snout-beetle Gonipterus scutellatus Gyllenhal (Coleoptera: Curculionidae) under laboratory conditions. By contact, entomopathogenic fungus B. bassiana EC and the pyrethroid λ-cyhalothrin exhibited the highest efficiency, achieving 100 and 97.5% mortality, respectively. By ingestion, the highest mortality was obtained by B. bassiana EC (100%) and thiamethoxam (95%). Flufenoxuron and lufenuron, bacteria mixture and M. anisopliae showed a weak toxicity. Furthermore, we found a sex-biased mortality, being males more affected. Due to the good performance and low risk to humans and environment, B. bassiana EC (strain PPRI 5339) appears to be the most promising product to promote an IPM programme in South Africa.     

Successful <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of <Emphasis Type="Italic">Populus tomentosa</Emphasis> with apple <Emphasis Type="Italic">SPDS</Emphasis> gene

The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree species with spermidine synthase （SPDS） genes derived from an apple by an Agrobacterium-mediated method. Four transgenic clones were confu＇med by PCR and Southern blot analysis. As well, the expression of introduced SPDS genes was analyzed by real-time quantitative PCR.     

Fluorescein-labeled wheat germ agglutinin stains the pine wood nematode, <Emphasis Type="Italic">Bursaphelenchus </Emphasis><Emphasis Type="Italic">xylophilus</Emphasis>

Pine wilt disease, caused by the pine wood nematode (PWN, Bursaphelenchus xylophilus), is a major threat to pine forests throughout East Asia. Nonetheless, its mechanism of invasion has not yet been described in detail. To better understand the pathology of this disease, it is important to examine the distribution of PWNs within pine tissue during the course of disease development. We attempted to stain nematodes with fluorescein-conjugated wheat germ agglutinin (F-WGA) as a means to locate and track the spread of PWNs. Although PWNs proliferated on Botrytis cinerea fungus were successfully stained only on their vulvas and spicule holes, PWNs extracted from inoculated Pinus thunbergii seedlings were stained on their surface. Stainability, or the percentage of PWNs stained with F-WGA over more than half of their surface, was about 20% by 1 day after inoculation, but increased to 80% at 10 days. The stainability of PWNs extracted from a 5-cm main stem segment that included the inoculation site was less than that of PWNs extracted from other parts of the main stem farther away (i.e., those that had dispersed). These results suggest that stainability is related to dispersal activity in time. Thus, to raise the stainability of PWNs at shorter timeframes after inoculation, PWNs with more than 80% stainability were re-inoculated into pine seedlings. This resulted in more than 70% stainability from 1 to 6 days after inoculation. In F-WGA stained thin paraffin sections of pine tissue of re-inoculated seedlings, PWNs brightly fluoresced under epifluorescence and were easily detected against the dark background of pine tissue. This staining technique with F-WGA is an excellent tool for detecting PWNs in pine tissue.     

Insecticidal efficacy of <Emphasis Type="Italic">Isaria farinosa</Emphasis> in different life stages of <Emphasis Type="Italic">Pissodes punctatus</Emphasis> (Coleoptera: Curculionidae)

The insecticidal activity of an entomopathogenic fungus, Isaria farinosa on pine bark-weevil (Pissodes punctatus) at different life stages underwent laboratory testing. Larvae, pupae and adults of Pissodes punctatus were exposed to a range of concentrations of fungal conidia in suspension for 5 s; mean mortality and LC₅₀ values were calculated. The results showed I. farinosa can effectively infect larvae, pupae and adult pine bark-weevils, and the highest mean mortalities at each life stage can exceed 88%. The mean mortalities increased with higher concentrations of I. farinosa. Larvae were most susceptible with LC₅₀ 1.2 × 10⁶ conidia ml⁻¹ (15 days), pupae (30 days) exhibited the second largest effects, and adults (15 days) with LC₅₀ 1.72 × 10⁶ and 1.99 × 10⁶ conidia ml⁻¹, respectively, at end of the experiments. This suggests that I. farinosa could be a potential bio-control agent against the pine bark-weevil.     

Effectiveness of Neem (<Emphasis Type="Italic">Azadirachta indica</Emphasis>) insecticides against Brassica pod midge (<Emphasis Type="Italic">Dasineura brassicae</Emphasis> Winn<Emphasis Type="Italic">.</Emphasis>)

The efficiency of the botanical insecticide (BI) NeemAzal T/S (containing 1% Azadirachtin A), on the basis of azadirachtin applied in a dose of 20 g a.i. ha⁻¹ against Brassica pod midge (Dasineura brassicae), has been monitored for 4 years. The biological efficiency of BI was compared with the efficiency of some synthetic insecticides. It was ascertained that BI was very efficient in decreasing the number of damaged oilseed rape pods (ranging from 56.5 to 85.9% compared to untreated plants) and its efficiency was comparable with synthetic insecticides based on Chloronicotinyl (Thiacloprid) and Neonicotinoid (Acetamiprid). BI’s efficiency was, in some years, even significantly higher compared to pyrethroid (λ-cyhalothrin). The high biological efficiency of azadirachtin that we ascertained was significantly reflected in increasing potential crop yields. The yield increase of azadirachtin ranged between 9.3 and 19.4% compared to the control sample. Azadirachtin showed the highest yield for the whole time of experimentation, and in some years the yield increase was significantly higher compared to some synthetic agents.     

Insecticidal properties of <Emphasis Type="Italic">Thymus persicus</Emphasis> essential oil against <Emphasis Type="Italic">Tribolium castaneum</Emphasis> and <Emphasis Type="Italic">Sitophilus oryzae</Emphasis>

Red flour beetle Tribolium castaneum (Herbst) and rice weevil Sitophilus oryzae (L.) are considered to be the major insect pests in storage. Essential oils from aromatic plants are recognized as proper alternatives to fumigants. Thymus persicus (Ronniger ex Rech. f.) is one of these plants that have medicinal properties and is indigenous to Iran. The essential oil was obtained from aerial parts of the plant and analyzed by GC and GC–MS. Carvacrol (44.69%) and thymol (11.05%) were the major constituents of the oil extracted. In this experiment, fumigant toxicity of the essential oil was studied against T. castaneum, S. oryzae at 27 ± 1°C and 60 ± 5% RH in dark condition. The adult insects were exposed to the concentrations of 51.9, 111.1, 207.4 and 370.4 μl/l air to estimate median lethal time (LT₅₀) values. The fumigant toxicity was increased in response to increased essential oil concentrations. The LT₅₀ values at the lowest and the highest concentrations tested were ranged from 28.09 to 13.47 h for T. castaneum, and 3.86 to 2.30 h for S. oryzae. It was found that S. oryzae adults were much more susceptible to the oil than T. castaneum. After 24 h of exposure, the LC₅₀ values (95% fiducial limit) for T. castaneum and S. oryzae were estimated to be 236.9 (186.27–292.81) and 3.34 (2.62–4.28) μl/l air, respectively. These results suggest that T. persicus essential oil merits further study as potential fumigant for the management of these stored-product insects.     

Side effects of selected insecticides on the <Emphasis Type="Italic">Tuta absoluta</Emphasis> (Lepidoptera: Gelechiidae) predators <Emphasis Type="Italic">Macrolophus pygmaeus</Emphasis> and <Emphasis Type="Italic">Nesidiocoris tenuis</Emphasis> (Hemiptera: Miridae)

Tuta absoluta is a pest native to South America that produces significant damage to tomato crops and was first detected in Europe in late 2006. Data obtained during 2008 from commercial tomato crops in which IPM was applied suggested that good pest control was possible through the combined action of the predatory mirid bugs Macrolophus pygmaeus and Nesidiocoris tenuis and the use of selective insecticides. The aim of our study was to evaluate the lethal and sublethal side effects of three of the insecticides most widely used to control T. absoluta on these predators: azadirachtin, spinosad and indoxacarb. Seven days after applying the treatment at the maximum recommended field rates, the mortality produced by indoxacarb ranged from 28% for nymphs of M. pygmaeus to 77% for females of N. tenuis and were significantly higher than those produced by azadirachtin, spinosad and the control (<13%). However, indoxacarb did not affect the number of descendants of females exposed to residues during the last days of their preimaginal development. In contrast, spinosad significantly reduced the offspring of M. pygmaeus and azadirachtin significantly reduced the offspring of N. tenuis females. The sublethal effects of azadirachtin and spinosad on predator reproduction should therefore not be ignored. This information could be useful when selecting the most appropriate insecticide to control T. absoluta in greenhouses and field crops in which M. pygmaeus and N. tenuis are used as biological control agents.     

Cultivation of <Emphasis Type="Italic">Pleurotus eryngii</Emphasis> on umbrella plant (<Emphasis Type="Italic">Cyperus alternifolius</Emphasis>) substrate

Mycelial growth and mushroom yields of three strains of Pleurotus eryngii produced on wheat bran-supplemented umbrella plant (Cyperus alternifolius) substrate were assessed using surface brightness, bromophenol blue color reactions, ergosterol and glucosamine contents, and water potential as indicators of strain performance. Mycelial growth was 31%–46% greater, depending on strain, on the umbrella plant substrate compared with the mushroom industry standard sugi (Cryptomeria japonica) substrate. Mushroom yields on the first flush were 20%–23% higher, depending on strain, on the plastic bottle-contained umbrella plant substrate. However, yields on the second break were lower from the umbrella plant substrate. Because many growers in Japan only harvest one flush, production of P. eryngii on umbrella plant substrate may offer commercial producers an alternative basal ingredient to diminishing supplies of sugi sawdust.Part of this report was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April 2002.     

Leaf-level responses to light in two co-occurring <Emphasis Type="Italic">Quercus</Emphasis> (<Emphasis Type="Italic">Quercus ilex</Emphasis> and <Emphasis Type="Italic">Quercus suber</Emphasis>): leaf structure,chemical composition and photosynthesis

We studied morphological, biochemical and physiological leaf acclimation to incident Photon-Photosynthetic-Flux-Density (PPFD) in Quercus ilex (holm oak) and Quercus suber (cork oak) at Mediterranean evergreen oak woodlands of southern Portugal. Specific leaf area (SLA) decreased exponentially with increasing PPFD in both species. Q. ilex had lower SLA values than Q. suber. Leaf nitrogen, cellulose and lignin concentration (leaf area-based) scaled positively with PPFD. Maximum rate of carboxylation (V_cmax), capacity for maximum photosynthetic electron transport (J_max), rate of triose-P utilization (V_TPU) and the rate of nonphotorespiratory light respiration (Rd) were also positively correlated with PPFD in both Quercus species, when expressed in leaf area but not on leaf mass basis. Q suber showed to have higher photosynthetic potential (V_cmax, J_max^m and V_TPU^m) and a higher nitrogen efficient nitrogen use than Q.ilex. Leaf chlorophyll concentration increased with decreasing PPFD, improving apparent quantum use efficiency (Φ) in both Quercus species. We concluded that, in Q.ilex and Q.suber, leaf structural plasticity is a stronger determinant for leaf acclimation to PPFD than biochemical and physiological plasticity.     

Pathogenic effects of <Emphasis Type="Italic">Xenorhabdus nematophilus</Emphasis> and <Emphasis Type="Italic">Photorhabdus luminescens</Emphasis> (Enterobacteriaceae) against pupae of the Diamondback Moth, <Emphasis Type="Italic">Plutella xylostella</Emphasis> (L.)

The relationship between Xenorhabdus nematophilus and Photorhabdus luminescens, the bacterial symbionts of the entomopathogenic nematodes Steinernema carpocapsae and Heterorhabditis bacteriophora, against the diamondback moth, P. xylostella (L.), pupae showed the pathogenic capability of P. luminescens to be over that of X. nematophilus. They gave 60 and 40?% mortality with LC₅₀ values of 5?×?10⁴ and 5.5?×?10⁵ cells/ml, respectively.The number of bacterial cells influences the rate of killing the pupae of P. xylostella and a direct correlation between mortality of the pupae, percentage of deformed adults and the spraying dose with significant differences were observed. An inverse correlation found between the no. of eggs laid/female, percentage of hatching, adult survival and the spraying dose. These bacterial effects have been attributed to different toxic bacterial enzymes which damage the haemocytes and inhibit activation of the humoral immune system of the insect.     

Comparison of postfire mortality in endemic Corsican black pine (<Emphasis Type="Italic">Pinus nigra</Emphasis> ssp. <Emphasis Type="Italic">laricio</Emphasis>) and its direct competitor (<Emphasis Type="Italic">Pinus pinaster</Emphasis>)

• Introduction   

Laricio pine (Pinus nigra J.F. Arn. ssp. laricio (Poiret) Maire var Corsicana Hyl.) is a form of black pine endemic to Corsica, that may now be under threat due to current fire regimes and competition with maritime pine (Pinus pinaster Aiton).     

Laboratory and field evaluation of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> for controlling Mulberry whitefly <Emphasis Type="Italic">Pealius mori</Emphasis> Takahashi (Homoptera: Aleyrodidae) in mulberry (<Emphasis Type="Italic">Morus alba</Emphasis> Linn)

In a laboratory screening of 12 isolates of entomopathogenic fungi against nymphs of the mulberry whitefly (Pealius mori Takahashi), Beauveria bassiana (Balsamo-Crivelli) Vuillemin CKB-048 was the most virulent, causing 87 ± 3% mortality at 1 × 10⁶ conidia/ml. Infection was confirmed by growth of the fungus from cadavers and by scanning electron microscopy of treated nymphs. Beauveria bassiana CKB-048 was formulated as a wettable powder (1 × 10⁹ conidia/g) and tested in two mulberry (Morus alba Linn) plantations in central and northeastern Thailand. In both locations, two spray applications of B. bassiana CKB-048 at 3.75 × 10¹² to 6.25 × 10¹² conidia/ha and at 14 day intervals provided good control of whitefly nymphs; control with B. bassiana CKB-048 was comparable to that with the pesticide buprofezin at 250 g of active ingredient/ha. In addition, no mortality of silkworm larvae occurred when the larvae were fed with mulberry leaves sprayed with B. bassiana CKB-048 7, 14, or 21 days earlier.