Non‐destructive observation of the mycangia of Euwallacea interjectus (Blandford) (Coleoptera: Curculionidae: Scolytinae) using X‐ray computed tomography

X‐ray microtomography has been applied successfully to obtain reliable microstructural information of many insect species. Nonetheless, the technique has not been widely applied to ambrosia beetles. The ambrosia beetle Euwallacea interjectus (Blandford) was first recorded as a vector of plant pathogenic fungus Ceratocystis ficicola Kajitani & Masuya, which has caused serious wilt disease in many fig orchards in Japan since 1999. Previous studies of E. interjectus have not described the mycangia (fungus‐storing organ) in detail. In this study, we non‐destructively examined the internal structure of an adult female of E. interjectus through computed microtomography scans. Paired mycangia were observed on typical computed tomography cross‐sections of the head. Each mycangium, ovoid in shape, was located in tissues just posterior to emarginated notch of eyes, adjacent to pharynx. Three dimensions (length × width × depth) of the mycangia were measured on stereography. We confirmed the absence of mycangia in the other body parts, such as elytra, prothorax and coxa of legs.     

Nematodes associated with Ips cembrae (Coleoptera: Curculionidae): comparison of generations,sexes and sampling methods

A study of nematodes associated with the large larch bark beetle Ips cembrae (Heer 1836) was carried out at three locations in the Czech Republic. The proportion of beetles infested by endoparasitic nematodes (representatives of genera Contortylenchus, Parasitylenchus, Cryptaphelenchus and Parasitorhabditis) ranged from 29.9 to 50.9%. Significant differences were determined in nematode infestation levels among locations, generations and sampling methods. No differences were found in infestation rates between males and females. The percentage of bark beetles with phoretic nematodes ranged from 18 to 42.9%. Phoretic nematodes directly found under elytra, on wings and between body segments of the bark beetles belong to the genus Micoletzkya. However, adults and juveniles of other two phoretic species Laimaphelenchus penardi and Bursaphelenchus sp. were found in the gallery frass of I. cembrae. Infestation by phoretic nematodes positively correlated with the presence of mites under elytra.     

External microstructure of the ambrosia beetle Platypus koryoensis (Coleoptera: Curculionidae: Platypodinae)

Platypus koryoensis is a minute ambrosia beetle found in forests. It can cause significant economic damage to oak trees. Recently in Korea, it has been reported as a major pest of oak trees, because it causes sooty mold of oak by introducing the pathogenic fungus Raffaelea sp. In this paper, we demonstrate the fine structural aspects of the external body of the ambrosia beetle using field emission scanning electron microscopy, as a part of basic research into this pest so that strategies for its control might be developed. This beetle has a sensory system well developed to respond to both visual and chemical stimuli. Both sexes have a pair of faceted compound eyes and a pair of knobbed antennae, but simple eyes are absent. The mouthparts on its distinct snouts are effective devices for penetration and for boring holes. The mouthparts consist of the labrum, a pair of mandibles, a pair of maxillae and the labium. Both the maxillary and the labial palpi have the function of directing the food to the mouth and holding it while the mandibles chew the food. The distal ends of these palpi are flattened and have shovel‐like setae. The thorax has a particularly hard exoskeleton and hard elytra, including powerful muscles that operate both the wings and the legs. The legs are multi‐segmented and have a strong femur and tibia, including one pair of claws on the end of each tarsal segment. Characteristically, both male and female beetles have mycangial cavities for storing spores and other microorganisms, but only females have three pairs of large depressions on their dorsal thorax.     

New host plant records for the Euwallacea fornicatus (Eichhoff) species complex (Coleoptera: Curculionidae: Scolytinae) across its natural and introduced distribution

Species within the Euwallacea fornicatus (Eichhoff) species complex are one of the few ambrosia beetles that infest healthy plants and are a potential phytosanitary threat as it causes considerable damage to many tree species in its native and introduced distribution. We updated the list to 412 plant species in 75 families that are known hosts for the E. fornicatus species complex, including 27 new host records, 20 of which are reproductive hosts, in the families Annonaceae, Apocynaceae, Araliaceae, Caricaceae, Euphorbiaceae, Fabaceae, Lauraceae, Magnoliaceae, Malvaceae, Meliaceae, Moraceae, Myristicaceae, Rubiaceae, Salicaceae, Sapindaceae, and Sapotaceae.     

Landscape‐scale genetic differentiation of a mycangial fungus associated with the ambrosia beetle,Xylosandrus germanus (Blandford) (Curculionidae:Scolytinae) in Japan

In this study, we examined the genetic structures of the ambrosia fungus isolated from mycangia of the scolytine beetle, Xylosandrus germanus to understand their co‐evolutionary relationships. We analyzed datasets of three ambrosia fungus loci (18S rDNA, 28S rDNA, and the β‐tubulin gene) and a X. germanus locus dataset (cytochrome c oxidase subunit 1 (COI) mitochondrial DNA). The ambrosia fungi were separated into three cultural morphptypes, and their haplotypes were distinguished by phylogenetic analysis on the basis of the three loci. The COI phylogenetic analysis revealed three distinct genetic lineages (clades A, B, and C) within X. germanus, each of which corresponded to specific ambrosia fungus cultural morphptypes. The fungal symbiont phylogeny was not concordant with that of the beetle. Our results suggest that X. germanus may be unable to exchange its mycangial fungi, but extraordinary horizontal transmission of symbiotic fungi between the beetle's lineages occurred at least once during the evolutionary history of this symbiosis.     

Nematodes associated with the double‐spined bark beetle Ips duplicatus (Coleoptera: Curculionidae) in central Europe

Ips duplicatus (Sahlberg) is an important forest pest in central Europe, but its nematode associates have seldom been studied. Therefore, nematodes associated with I. duplicatus were determined at three localities in the Czech Republic. The percentage of beetles with phoretic nematodes ranged from 18 to 65%. Micoletzkya buetschlii and other phoretic nematodes were found under elytra, on wings, and between body segments. The percentage of beetles with nematodes in the haemocoel ranged from 3 to 30%, and the nematodes included Contortylenchus diplogaster and Parasitylenchus cf. aculeatus. Juveniles of Parasitorhabditis obtusa (Fuchs 1915) were found in the intestines of 0–16% of the beetles. The most abundant species in I. duplicatus galleries were P. obtusa and M. buetschlii. Cryptaphelenchus sp., Parasitaphelenchus sp. and unidentified tylenchid juveniles also were found in the galleries. The percentage of beetles with nematodes was greater in the overwintering than in the offspring generation, and numbers of nematodes per gallery increased with gallery development.     

Local augmentation efficiency of Anaphes nitens (Hymenoptera,Mymaridae), the egg parasitoid of Gonipterus platensis (Coleoptera,Curculionidae)

The efficiency of local augmentation releases of the egg parasitoid Anaphes nitens to control the Eucalyptus snout‐beetle Gonipterus platensis was tested in Eucalyptus globulus plantations in Galicia (NW Spain). On May–June 2006, at two localities of Pontevedra province, the release of host egg capsules parasitized by A. nitens at a potential rate of 300 adults/ha was compared with a release density of 900 adults/ha, and a control group of eucalypts not subjected to augmentation. Parasitism rate after 1–2 weeks did not significantly increase over the control plots at both localities. The high release rate did not ensure a higher crop protection and therefore could be not economically justified. On March–April 2017, at four localities of Pontevedra province, the test was replicated by releasing 300 parasitoids/ha. Parasitism level did significantly increase over the control just in one locality. Augmentation of A. nitens at small scale generally failed to achieve a higher protection from the pest, possibly due to the large extension of the E. globulus plantations, the magnitude of the G. platensis population and the fluctuations of the established parasitoid population, whose density is in turn affected by host egg availability and density‐dependent dispersal.     

The Banded Elm Bark Beetle, Scolytus schevyrewi Semenov (Coleoptera, Curculionidae, Scolytinae) in North America: a taxonomic review and modifications to the Wood (1982) key to the species of Scolytus Geoffroy in North and Central America

In 2003, an Asian bark beetle, Scolytus schevyrewi Semenov (Coleoptera: Curculionidae: Scolytinae), the banded elm bark beetle, was detected for the first time in North America. This paper modifies the Wood (1982) key to the species of Scolytus Geoffroy to enable identification of Scolytus schevyrewi in North and Central America. Variation of diagnostic characters in Scolytus schevyrewi is discussed.     

Dispersal behaviour of Euwallacea nr. fornicatus (Coleoptera: Curculionidae: Scolytinae) in avocado groves and estimation of lure sampling range

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Areography of the genus Dendroctonus (Coleoptera: Curculionidae: Scolytinae) in Mexico

Objective To analyse whether the geographical ranges of Dendroctonus species are (1) associated with factors such as host species or elevation, and (2) in agreement with Halffter's Nearctic distribution pattern. (3) To identify and discuss the factors that are likely to act as barriers to the genus’ geographical distribution. (4) To explore whether there is an association between the size of the geographical ranges of Dendroctonus species and the number of Pinus host species used by each of them, and (5) to assess if these host species are most common at the elevations preferred by the individual Dendroctonus species. Site Mexico. Methods Records of 12 species of Dendroctonus were gathered from entomological collections in Mexico. Distribution ranges were defined by using the propinquity method ( Rapoport, 1975a ). Analysed parameters were: (1) geographical distribution of single species, (2) overlapping of species ranges, (3) disjunction patterns and barriers by means of isoprobabilistic lines, based on the morphotectonic subdivision of Mexico ( Ferrusquía‐Villafranca, 1998 ), (4) spatial variation in species richness with respect to latitude and altitude, (5) size of geographical ranges, and (6) host species for each Dendroctonus species. A correlation was determined between area size and number of pine host species. Results The species ranges varied in shape and size. Geographical ranges tend to be discontinuous in shape. Composite patterns showed that disjunctions among ranges do not closely follow Mexico's morphotectonic subdivision. There are repeated discontinuities among individual distributions, which define five areas: (1) Baja California Peninsula, (2) Sierra Madre Occidental (SMOC), (3) northern Sierra Madre Oriental (SMOR), (4) Sierra Madre de Chiapas, and (5) SMOR + Faja Volcanica Transmexicana (FVT) + Sierra Madre del Sur. The isoprobabilistic lines confirm that the inner part of SMOC provides an optimal environment for the genus, and the FVT province constitutes the broader corridor for it in the country. Richness does not directly decrease or increase with latitude. Richness behaviour of the insect is not associated with that of its host. Elevation distributions showed that most Dendroctonus species move within broad margins of tolerance and species richness is concentrated in the montane interval. Dendroctonus attack 24 of the 47 Pinus species distributed in Mexico. Preferred pine species belong predominantly to Leiophyllae, Ponderosae and Oocarpae subsections. The Spearman rank correlation between area size and number of pine host species was not significant. Dendroctonus clearly belongs to a Nearctic distribution pattern (sensu Halffter, 1987 ). Main conclusions Dendroctonus is present in all montane systems of Mexico and its species coexist within a high geographical sympatry. Overlapping of species distribution appears to be the result of two elements – generalized polyphagy inside Pinus and a wide elevation tolerance within mountainous environments. This behaviour, linked to a high vagility, has allowed the genus Dendroctonus to expand its distribution across Mexico and to employ mountainous systems as corridors separated by barriers that exert a low selective filter effect.     

Effectiveness of different lures for attracting Ips acuminatus (Coleoptera: Curculionidae: Scolytinae)

1. Ips acuminatus (the sharp-toothed bark beetle, STBB) is currently considered to be one of the most serious pests of Scots pine in many European countries. STBB management is among the most challenging tasks in pine forests; the development of methods for monitoring, predicting and managing outbreaks of this bark beetle is therefore crucial.
2. Pheromone-baited traps have been widely recommended as a valuable tool for the monitoring and mass trapping of bark beetles. Although different suppliers offer a variety of STBB lures, their effectiveness has rarely, if ever, been evaluated under natural conditions.
3. We evaluated the attractiveness of three commercially available and five experimental synthetic lures by comparing the numbers of STBBs captured in white, six-funnel traps. The studies were conducted in 2017–2019 in Poland, in Scots pine-dominated forests affected by STBB outbreaks.
4. Our study demonstrated significant differences in the effectiveness of the lures. The experimental lure produced by the Witasek company (Austria) and the recently marketed lure Acumodor Micro from Chemipan (Poland) were the most attractive to STBB. Among the least effective were two commercial lures (Acuwit and Acumodor), hitherto used in Central Europe.
5. The results will be useful in developing methods for the monitoring and management of STBB populations.

     

Tracing the origin of a cryptic invader: phylogeography of the Euwallacea fornicatus (Coleoptera: Curculionidae: Scolytinae) species complex

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Genetic structure of Japanese populations of an ambrosia beetle,Xylosandrus germanus (Curculionidae: Scolytinae)

We examined the genetic structures of 13 Japanese populations of an ambrosia beetle, Xylosandrus germanus (Curculionidae: Scolytinae), to understand the effects of geographical barriers on the colonization dynamics of this species. The genetic structure was studied using portions of the mitochondrial cytochrome oxidase I (COI) gene. A phylogenetic analysis revealed three distinct lineages (clades A, B and C) within X. germanus. Clade A contained 21 haplotypes from all 13 populations; whereas clade B contained eight haplotypes from Hokkaido (Sapporo and Furano), Iwate and Nagano populations; and clade C contained only a single a haplotype from the Hokkaido (Furano) population. In the analysis of molecular variance (amova ), the greatest amount of genetic variation was detected between populations in Hokkaido and those in Honshu and other southern islands. Between these two groups of populations, all the values of the coefficient of gene differentiation were significantly larger than zero, except for the Hokkaido (Sapporo) versus Nagano comparison. Our results confirm that for X. germanus, gene flow has been interrupted between Hokkaido and Honshu since the last glacial maximum.     

Verbenone-releasing flakes protect individual Pinus contorta trees from attack by Dendroctonus ponderosae and Dendroctonus valens (Coleoptera: Curculionidae, Scolytinae)

1 In a study site in interior northern California, twenty individual lodgepole pines Pinus contorta were sprayed with a suspension of DISRUPT Micro‐Flake^® Verbenone (4,6,6‐trimethylbicyclo(3.1)hept‐3‐en‐2‐one) Bark Beetle Anti‐Aggregant flakes (Hercon Environmental, Emigsville, Pennsylvania) in water, with sticker and thickener, from ground level to a height of 7 m. Twenty trees sprayed with just water, sticker and thickener served as controls. All trees were baited immediately after spraying with mountain pine beetle Dendroctonus ponderosae aggregation pheromone lures, and lures were refreshed after 4 weeks. 2 Trees treated with verbenone had significantly lower attack density by D. ponderosae than controls at 2, 4, 6 and 8 weeks after application of flakes. 3 None of the treated trees was attacked by red turpentine beetle Dendroctonus valens, whereas control trees averaged nearly two D. valens attacks per tree, 8 weeks after treatment. 4 A dry frass index, used to predict ultimate tree mortality, was significantly higher in control trees than treated trees for all four sampling intervals. This index proved to be a significant predictor of ultimate tree mortality. 5 Ten months after application, treated trees showed significantly lower mortality than control trees.     

Real‐time PCR identification of the ambrosia beetles,Trypodendron domesticum (L.) and Trypodendron lineatum (Olivier) (Coleoptera: Scolytidae)

The European hardwood ambrosia beetle (Trypodendron domesticum) and the striped ambrosia beetle (Trypodendron lineatum) are wood‐boring pests that can cause serious damage to lumber, resulting in a need for management of these pests in logging and lumber industries. Natural populations of ambrosia beetles exist throughout the world, but movement of ambrosia beetles into new habitats, particularly via international trade, can result in the establishment of invasive species that have the potential to spread into new territory. Efforts to monitor ambrosia beetle populations are time‐consuming and could be greatly enhanced by the use of molecular methods, which would provide accurate and rapid identification of potentially problematic species. Here, we present new real‐time PCR assays for the detection and identification of T. domesticum and T. lineatum. The methods described herein can be used with a variety of sampling strategies to enable timely and well‐informed decision‐making in efforts to control these ambrosia beetles.     

DNA条形码试剂盒检测技术在大小蠹属种类鉴定中的应用

[目的]DNA条形码技术已成为生物分类鉴定的有力工具.DNA条形码技术的相关问题,如物种种内和种间的遗传距离出现重叠区域,将直接影响到物种鉴定的准确性.我们应用DNA条形码试剂盒检测技术来快速、准确地鉴定口岸截获的检疫性大小蠹属种类.[方法]针对大小蠹昆虫设计引物以提高PCR扩增效率.运用自主研发的基因条码分析软件找出基因片段上区分每个物种的多态位点规律,作为该物种的鉴定特征并建立数据库,应用于物种鉴定.[结果]使用针对大小蠹属昆虫设计的引物成功扩增出325 bp的COI基因片段.将大小蠹属12种昆虫的COI基因片段上的核苷酸诊断位点的组合作为物种的鉴定特征,可以准确地区分近似种.通过比对植物检疫鉴定系统数据库里的鉴定特征,将6个大小蠹属的未知样品成功鉴定到种(核苷酸序列一致性为100％),与形态鉴定结果一致.[结论]结果表明DNA条形码试剂盒检测技术可以准确鉴定大小蠹属的种类.该检测技术可以应用于其他经济重要性有害生物的检测鉴定.     

Effect of associated fungi on the immunocompetence of red turpentine beetle larvae,Dendroctonus valens (Coleoptera: Curculionidae: Scolytinae)

Abstract Dendroctonus–fungus symbioses are often considered as the ideal model systems to study the development and maintenance of ectosymbioses, and diverse interactions, including antagonism, commensalism and mutualism, have been documented between these organisms. The red turpentine beetle, Dendroctonus valens LeConte (Coleoptera: Curculionidae: Scolytinae) is a pine‐killing invasive beetle in northern China. Fungi species Ophiostoma minus, Leptographium sinoprocerum, L. terebrantis and L. procerum were associated with this bark beetle. Antagonistic interactions between D. valens and its associated fungi, such as O. minus and L. sinoprocerum, have been demonstrated, but the underlying causes of this phenomenon are unknown. Here, we first found the two tested fungi species retarded the net weight gain of D. valens larvae after completing 3‐day feeding on their media. Furthermore, we provide direct evidence indicating the effect of associated fungi on the immunocompetence of D. valens larvae to explain the documented antagonism. Our results showed that the activity of phenoloxidase and total phenoloxidase in D. valens larvae were significantly upregulated by two strains of associated fungi, O. minus and L. sinoprocerum as compared with the controls. The phenoloxidase ratio increased significantly in the larvae which had fed for 3 days on media inoculated with O. minus. Because insect immune defenses are costly to be deployed, these results could be explored as one of the underlying mechanisms of the documented antagonism.     

Comparative morphology of sensilla on antenna,maxillary palp and labial palp of larvae of Eucryptorrhynchus scrobiculatus (Olivier) and E. brandti (Harold) (Coleoptera: Curculionidae)

Eucryptorrhynchus scrobiculatus (Olivier) and E. brandti (Harold) are two wood boring pests of Ailanthus altissima (Mill.) Swingle (tree of heaven) and the variety Ailanthus altissima var. Qiantouchun. These beetles attack healthy trees and bore into roots and trunks during the larval stage. We studied the typology, distribution and morphostructure of the sensilla on the antennae, maxillary palps and labial palps of E. scrobiculatus and E. brandti larvae using scanning and transmission electron microscopy. The results showed the following: (i) the antennae of the two weevil larvae had two types of sensilla, sensilla basiconica (S.b.1 and S.b.2) and sensilla twig basiconica (S.tb.1‐S.tb.3), with S.tb.4 observed only on the antennae of E. brandti larvae; (ii) the maxillary palps had three types of sensilla, S.b.2, S.tb. (S.tb.2, S.tb.3 and S.tb.5) and digitiform sensilla; (iii) the labial palps had two types of sensilla, S.b.2 and S.tb. (S.tb.2, S.tb.3 and S.tb.5); (iv) the quantity and distribution of sensilla on the antennae, maxillary palps and labial palps remained constant between E. scrobiculatus and E. brandti larvae; and (v) sensilla basiconica had distinct sidewall pores, an apical pore was observed on sensilla twig basiconica, and digitiform sensilla were oval in shape, with a distinct apical pore. Based on the microstructure of the cuticle wall and dendrite, we hypothesized that these sensilla functioned as olfactory, gustatory and hygro‐/thermo‐receptors, respectively. We discuss the relationships among types of sensilla and the types of damage caused by the larvae inside the host tree to understand olfactory and gustatory receptor mechanisms. The results of this study will provide a firm basis for future electrophysiological studies.     

Molecular characterization,virulence and horizontal transmission of Beauveria pseudobassiana from Dendroctonus micans (Kug.) (Coleoptera: Curculionidae)

The great spruce bark beetle, Dendroctonus micans (Kugelann) (Coleoptera: Curculionidae), has been a potential threat for Turkey and the entire Eurasian spruce forests for many years. Control strategies which have been applied so far are still insufficient to prevent its damage. A previous study has shown that a Beauveria isolate (ARSEF 9271) proved to be an efficient microbial control agent against the great spruce bark beetle. In this study, this isolate was identified as B. pseudobassiana based on the partial sequence of EF1‐α and ITS sequence. A conidial suspension (1 × 10⁸/ml) of this fungus caused 100% mortality on both larvae and adults of D. micans within 5 and 6 days, respectively. Also, it caused 100% mycosis value on both larvae and adults. Mortality values of horizontal transmission experiments between larvae and adults which were contaminated with 1 × 10⁶/ml spore suspension at 25%, 50%, 75% and 100% rates were determined as 100% after 15 days at 20°C under the laboratory conditions. We also determined the decrease of the damage in spruce wood block (15 × 25 cm) when the contamination rate of the larvae was increased. Our results indicate that B. pseudobassiana ARSEF 9271 seems to be a very promising biocontrol agent against D. micans.