The diversity and abundance of wild bees ensures the delivery of pollination services and the maintenance of ecosystem diversity. As previous studies carried out in Central Europe and the US have shown, bee diversity and abundance is influenced by the structure and the composition of the surrounding landscape. Comparable studies have so far not been carried out in the Mediterranean region. The present study examines the influence of Mediterranean landscape context on the diversity and abundance of wild bees. To do this, we sampled bees in 13 sites in olive groves on Lesvos Island, Greece. Bees were assigned to five categories consisting of three body size groups (small, medium and large bees), the single most abundant bee species (Lasioglossum marginatum) and all species combined. The influence of the landscape context on bee abundance and species richness was assessed at five radii (250, 500, 750, 1000 and 1250 m) from the centre of each site. We found that the abundance within bee groups was influenced differently by different landscape parameters and land covers, whereas species richness was unaffected. Generally, smaller bees’ abundance was impacted by landscape parameters at smaller scales and larger bees at larger scales. The land cover that influenced bee abundance positively was olive grove, while phrygana, conifer forest, broad-leaved forest, cultivated land, rock, urban areas and sea had mostly negative or no impact. We stress the need for a holistic approach, including all land covers, when assessing the effects of landscape context on bee diversity and abundance in the Mediterranean.

Much of the ecological alteration faced by human-modified Neotropical forests can be assigned to edge effects, including the proliferation of some voracious herbivores such as leaf-cutting ants. However, the underlying mechanisms/impacts of tropical forest edge on herbivores performance and their foraging behaviour (e.g. dietary diversity) have rarely been investigated. The goal of this study was, therefore, to determine whether and how the annual diet (i.e. species richness, diversity and the relative proportion of pioneer versus non-pioneer species of plant materials) of Atta cephalotes colonies differs in the forest edge versus the interior zone of a large remnant of Atlantic forest in northeastern Brazil. Among the key results was a strong habitat effect on dietary diversity (explaining ca. 40–50% of the variation), which, in edge colonies, decreased approximately by one fourth compared to interior colonies (inverse of Simpson's index: 3.7±0.84 versus 4.99±0.95). There was a predominance of leaf fragments collected from pioneer species in the diet in both habitat (86% in edge and 80.4% in interior). Edge colonies collected proportionally more fragments from pioneer species than colonies located in the forest interior. Our results are the first to demonstrate an edge-mediated relaxation of dietary restrictions in leaf-cutting ants. These findings render robust support to previous evidence indicating the reduction of bottom-up forces as a key factor explaining both edge-induced hyper-abundance and increased herbivory of leaf-cutting ants in human-modified Neotropical landscapes.

Native insects can become epidemic pests in agro-ecosystems. A population genetics approach was applied to analyze the emergence and spread of outbreak populations of native insect species. Outbreaks of the mirid bug, Stenotus rubrovittatus, have rapidly expanded over Japan within the last two decades. To characterize the outbreak dynamics of this species, the genetic structure of local populations was assessed using polymorphisms of the mtDNA COI gene and six microsatellite loci. Results of the population genetic analysis suggested that S. rubrovittatus populations throughout Japan were genetically isolated by geographic distance and separated into three genetic clusters occupying spatially segregated regions. Phylogeographic analysis indicated that the genetic structure of S. rubrovittatus reflected post-glacial colonization. Early outbreaks of S. rubrovittatus in the 1980s occurred independently of genetically isolated populations. The genetic structure of the populations did not fit the pattern of an outbreak expansion, and therefore the data did not support the hypothesis that extensive outbreaks were caused by the dispersal of specific pestiferous populations. Rather, the historical genetic structure prior to the outbreaks was maintained throughout the increase in abundance of the mirid bug. Our study indicated that changes in the agro-environment induced multiple outbreaks of native pest populations. This implies that, given suitable environmental conditions, local populations may have the potential to outbreak even without invasion of populations from other environmentally degraded areas.

Click beetle larvae within the genus Agriotes (Coleoptera: Elateridae), commonly known as wireworms, are abundant ground-dwelling herbivores which can inflict considerable damage to field crops. In Central Europe up to 20 species, which differ in their distribution, ecology and pest status, occur in arable land. However, the identification of these larvae based on morphological characters is difficult or impossible. This hampers progress towards controlling these pests. Here, we present a polymerase chain reaction (PCR)-based approach to identify, for the first time, 17 Agriotes species typically found in Central Europe. Diagnostic sequence information was generated and submitted to GenBank, allowing the identification of these species via DNA barcoding. Moreover, multiplex PCR assays were developed to identify the nine most abundant species rapidly within a single-step reaction: Agriotes brevis, A. litigiosus, A. obscurus, A. rufipalpis, A. sordidus, A. sputator, A. ustulatus, A. lineatus and A. proximus. The latter two species remain molecularly indistinguishable, questioning their species status. The multiplex PCR assays proved to be highly specific against non-agrioted elaterid beetles and other non-target soil invertebrates. By testing the molecular identification system with over 900 field-collected larvae, our protocol proved to be a reliable, cheap and quick method to routinely identify Central European Agriotes species.

Insecticide resistance is an important factor in the effectiveness of Aedes aegypti control and the related spread of dengue. The objectives of this study were to investigate the status of the organochlorine dichlorodiphenyltrichloroethane (DDT) and pyrethroid (permethrin and deltamethrin) resistance in Trinidad and Tobago populations of Ae. aegypti and the underlying biochemical mechanisms. Nine populations of Ae. aegypti larvae from Trinidad and Tobago were assayed to DDT and PYs using the Centers for Disease Control and Prevention (CDC) time-mortality-based bioassay method. A diagnostic dosage (DD) was established for each insecticide using the CAREC reference susceptible Ae. aegypti strain and a resistance threshold (RT), time in which 98–100% mortality was observed in the CAREC strain, was calculated for each insecticide. Mosquitoes which survived the DD and RT were considered as resistant, and the resistance status of each population was categorised based on the WHO criteria with mortality <80% indicative of resistance. Biochemical assays were conducted to determine the activities of α and β esterases, mixed function oxidases (MFO) and glutathione-S-transferases (GST) enzymes which are involved in resistance of mosquitoes to DDT and PYs. Enzymatic activity levels in each population were compared with those obtained for the CAREC susceptible strain, and significant differences were determined by Kruskal-Wallis and Tukey's non-parametric tests (P<0.05). The established DDs were 0.01 mg l−1, 0.2 mg l−1 and 1.0 mg l−1 for deltamethrin, permethrin and DDT, respectively; and the RTs for deltamethrin, permethrin and DDT were 30, 75 and 120 min, respectively. All Ae. aegypti populations were resistant to DDT (<80% mortality); two strains were incipiently resistant to deltamethrin and three to permethrin (80–98% mortality). Biochemical assays revealed elevated levels of α-esterase and MFO enzymes in all Ae. aegypti populations. All, except three populations, showed increased levels of β-esterases; and all populations, except Curepe, demonstrated elevated GST levels.

Metabolic detoxification of enzymes is correlated with the manifestation of DDT and PY resistance in Trinidad and Tobago populations of Ae. aegypti. The presence of this resistance also suggests that knock down (kdr)-type resistance may be involved, hence the need for further investigations. This information can contribute to the development of an insecticide resistance surveillance programme and improvement of resistance management strategies aimed at combatting the spread of dengue in Trinidad and Tobago.

Soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), is currently the most important insect pest of soybean (Glycine max (L.) Merr.) in the United States and causes significant economic damage worldwide, but little is known about the aphid at the molecular level. Mariner-like transposable elements (MLEs) are ubiquitous within the genomes of arthropods and various other invertebrates. In this study, we report the cloning of MLEs from the soybean aphid genome using degenerate PCR primers designed to amplify conserved regions of mariner transposases. Two of the ten sequenced clones (designated as Agmar1 and Agmar2) contained partial but continuous open reading frames, which shared high levels of homology at the protein level with other mariner transposases from insects and other taxa. Phylogenetic analysis revealed Agmar1 to group within the irritans subfamily of MLEs and Agmar2 within the mellifera subfamily. Southern blot analysis and quantitative PCR analysis indicated a low copy number for Agmar1-like elements within the soybean aphid genome. These results suggest the presence of at least two different putative mariner-like transposases encoded by the soybean aphid genome. Both Agmar1 and Agmar2 could play influential roles in the architecture of the soybean aphid genome. Transposable elements are also thought to potentially mediate resistance in insects through changes in gene amplification and mutations in coding sequences. Finally, Agmar1 and Agmar2 may represent useful genetic tools and provide insights on A. glycines adaptation.

The effect of four temperatures (18, 20, 25 and 30°C) on pupa development and sexual maturity of Anastrepha obliqua adults was investigated under laboratory conditions. The results showed that the duration of the pupal stage decreased with an increase in temperature (29, 25, 13 and 12 days, respectively), and maintaining the pupae at 18°C and 20°C results in a low percentage of pupation, pupa weight loss and lesser flying ability. However, it significantly favored sexual behavior, a higher proportion of sexual calls and matings. While enhanced pupa development was observed at a temperature of 30°C, adults had low sexual efficiency, as well as a lower proportion of calls and matings. Gas chromatography-mass spectrometry (GC-MS) analysis of male volatiles showed that the amount of (Z,E)-α-farnesene did not vary among males from pupae reared at different temperatures; however, less (E,E)-α-farnesene was emitted by males obtain from pupa reared at 30°C. Male flies kept at 30°C during their larval stage had more (Z)-3-nonenol and, also, an unknown compound was detected. The fecundity of the females was higher at low temperatures. Regarding fertility, no significant differences were found between temperatures. The optimal temperature on pupa development was 25°C when males displayed ideal attributes for rearing purposes.

Two novel full-length P450 cDNAs, CYP6CS1 and CYP6CW1, were cloned from the fourth instar nymphs of brown planthopper Nilaparvata lugens Stål (Hemiptera: Delphacidae) reared on its susceptible rice variety Taichung Native 1 (TN1) plants. The deduced proteins are typical microsomal P450s sharing conserved structural and functional domains with other insect CYP6 members. Temporal expression analysis by northern blot hybridization indicated pre-exposure to N. lugens moderately resistant rice Minghui 63 (MH63) seedlings caused a time course-dependent induction of CYP6CS1 which peaked after 24 h of treatment; in contrast, CYP6CW1 was induced and remained at a constant time course from 0–72 h. CYP6CS1 and CYP6CW1 are dramatically induced in gut tissues and, slightly upregulated in carcass and fat bodies as revealed in spatial gene expression analysis. Whole mount in situ hybridizaion revealed that the two genes are expressed at a basal level in gut tissue and Malpighian tubules in nymphs fed with TN1 rice. After exposure to MH63, the expression of CYP6CW1 was found to be high in the whole gut, including Malpighian tubules. Expression of CYP6CS1 was significantly increased in midgut, and slightly increased in foregut, hindgut and Malpighian tubules. These data suggest a potential role of the two P450s in determining patterns of N. lugens-rice relationships through allelochemical detoxification.

Effects of host species, stage and size on clutch size and sex ratio of the gregarious, idiobiont ectoparasitoid Dibrachys boarmiae were investigated at 25±2°C and 70±5% relative humidity. The greater wax moth, Galleria mellonella, small wax moth, Achroia grisella, and early stage solitary larvae of the endoparasitoid, Apanteles galleriae, were used as hosts. Clutch size was greatest from prepupae of the largest host, Galleria mellonella, with a mean of 40.07 offspring per host versus 14.73 and 2.93 for Achroia grisella and Apanteles galleriae, respectively. The mean clutch size from pupae was lower than from prepupae, being 17.27, 10.73 and 2.89 for Galleria mellonella, Achroia grisella and Apanteles galleriae, respectively. Within each host species and stage, heavier hosts resulted in larger clutches. The sex ratio of offspring (proportion of male) was approximately 0.20, with only minor differences among host species, stages and sizes.

The tobacco whitefly, Bemisia tabaci (Gennadius), causes severe crop losses in many agricultural systems. The worst of these losses are often associated with the invasion and establishment of specific whitefly biotypes. In a comprehensive survey of biotypes present in central China between 2005 and 2007, we obtained 191 samples of B. tabaci from 19 districts in Hubei province and its surrounds. Biotypes were identified by RAPD-PCR and by sequencing the mitochondrial cytochrome oxidase I gene (mtCO1). We determined that these central Chinese haplotypes included the world's two most invasive B. tabaci biotypes (B and Q) and two indigenous biotypes (ZHJ1 and ZHJ3). The B biotype shared >99.7% identity with other Chinese B biotypes and the Q biotype shared >99.5% of its identity with Q samples from the Mediterranean, USA, Africa and East Asia. By 2007, the Q biotype was dominant over much of Hubei province and appeared to be supplanting all other biotypes, although both the invasive and indigenous biotypes existed in sympatry in some regions. The invasion and rapid establishment of the Q biotype in China mirrors events elsewhere in the world, and we suggest that this is a consequence of its reproductive isolation, its polyphagous nature and its broad-spectrum resistance to insecticides. Its dominance has severe implications for the sustainability of some insecticide groups and for the production of a number of crops.

We, herein, report evidence that wing fanning by the aphid parasitoid Aphidius ervi Haliday (Hymenoptera: Braconidae) constitutes a courtship song. Complete removal of the forewings or only the distal half of them reduced male copulation success in comparison to intact males. Males that achieved copulation within the observation period produced wing fanning at a higher rate than males that did not copulate. Playback of wing fanning sound altered the behaviour of virgin females, increasing the time they devoted to grooming, as compared with subjects that were exposed to silence or white noise. Our findings contribute to the understanding of the acoustic dimension of the sensory modalities employed by this aphid parasitoid in sexual signalling.

Polyacrylamide gel electrophoresis of enzymes (carboxylesterases) was used for the first time to monitor rates of parasitism in airborne alate (winged) grain aphid, Sitobion avenae (F.) population samples collected by suction trapping in Hertfordshire, UK. Using previously described electrophoretic ‘keys’, the species of hymenopterous parasitoids present in individual aphids were identified and found to be Aphidius ervi (Haliday) and/or Aphidius rhopalosiphi (De Stephani Perez) (Braconidae). Entomophthoralean fungal infection was also detected using this approach. Aphidiid wasp parasitism was detected from early June to mid-August and fungal infection from late June to late July. The results are discussed in relation to parasitoid population structure and dynamics, especially (i) the fact that winged aphids passively transport the early stages of their braconid parasitoids and fungal pathogens, potentially to newly-founded colonies, which may directly impact on the dual aphid-parasitoid populations genetics; and (ii) the approach used to collect and assay parasitised and fungal infected aphids involving both suction trapping and electrophoretic testing may have potential in assessing the level and efficacy of these biological control agents in integrated pest management (IPM) schemes to combat cereal aphid outbreaks.

Population cycles of the winter moth (Operophtera brumata) in sub-arctic coastal birch forests show high spatiotemporal variation in amplitude. Peak larval densities range from levels causing little foliage damage to outbreaks causing spatially extensive defoliation. Moreover, outbreaks typically occur at or near the altitudinal treeline. It has been hypothesized that spatiotemporal variation in O. brumata cycle amplitude results from climate-induced variation in the degree of phenological matching between trophic levels, possibly between moth larvae and parasitoids. The likelihood of mismatching phenologies between larvae and parasitoids is expected to depend on how specialized parasitoids are, both as individual species and as a guild, to attacking specific larval developmental stages (i.e. instars). To investigate the larval instar-specificity of parasitoids, we studied the timing of parasitoid attacks relative to larval phenology. We employed an observational study design, with sequential sampling over the larval period, along an altitudinal gradient harbouring a pronounced treeline outbreak of O. brumata. Within the larval parasitoid guild, containing seven species groups, the timing of attack by different groups followed a successional sequence throughout the moth's larval period and each group attacked 1–2 instars. Such phenological diversity within parasitoid guilds may lower the likelihood of climate-induced trophic mismatches between victim populations and many/all of their enemies. Parasitism rates declined with increasing altitude for most parasitoid groups and for the parasitoid guild as a whole. However, the observed spatiotemporal parasitism patterns provided no clear evidence for or against altitudinal mismatch between larval and parasitoid phenology.

The B and Q ‘biotypes’ of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) have been invading many parts of the world and causing severe damage to a range of crops. Recent phylogenetic analyses indicate that B and Q are cryptic species within the B. tabaci species complex. Although various attempts have been made to examine the reproductive compatibility between B and Q, few studies have tested the fertility of the F1 females and so the extent of possible gene flow remains unclear. In this study, we conducted a series of crossing experiments and behavioural observations to examine in detail the reproductive compatibility between the B and Q biotypes collected from Zhejiang, China, a region recently invaded by these whiteflies. Crossing experiments between the two biotypes using either single-pairs or small groups demonstrated that proportions of females in the F1 progeny were only 0–2% in the inter-biotype crosses compared to 58–68% in the intra-biotype treatments. Furthermore, all inter-biotype F1 females were sterile. Continuous video observations showed that B and Q adults very rarely copulated, and copulation occurred only when adults of opposite sex from different biotypes were enclosed in dense cohorts for a relatively long period of time. These data show that the B and Q biotypes examined in this study are completely isolated in reproduction. The isolation was due to mainly a copulation barrier, but post-copulation barriers were also involved.

The Asian citrus psyllid, Diaphorina citri Kuwayama, vectors Candidatus Liberibacter asiaticus (Las) and Candidatus Liberibacter americanus (Lam), the presumed causal agents of huanglongbing. D. citri generally rely on olfaction and vision for detection of host cues. Plant volatiles from Allium spp. (Alliaceae) are known to repel several arthropod species. We examined the effect of garlic chive (A. tuberosum Rottl.) and wild onion (A. canadense L.) volatiles on D. citri behaviour in a two-port divided T-olfactometer. Citrus leaf volatiles attracted significantly more D. citri adults than clean air. Volatiles from crushed garlic chive leaves, garlic chive essential oil, garlic chive plants, wild onion plants and crushed wild onion leaves all repelled D. citri adults when compared with clean air, with the first two being significantly more repellent than the others. However, when tested with citrus volatiles, only crushed garlic chive leaves and garlic chive essential oil were repellent, and crushed wild onions leaves were not.

Analysis of the headspace components of crushed garlic chive leaves and garlic chive essential oil by gas chromatography-mass spectrometry revealed that monosulfides, disulfides and trisulfides were the primary sulfur volatiles present. In general, trisulfides (dimethyl trisulfide) inhibited the response of D. citri to citrus volatiles more than disulfides (dimethyl disulfide, allyl methyl disulfide, allyl disulfide). Monosulfides did not affect the behaviour of D. citri adults. A blend of dimethyl trisulfide and dimethyl disulfide in 1:1 ratio showed an additive effect on inhibition of D. citri response to citrus volatiles. The plant volatiles from Allium spp. did not affect the behaviour of the D. citri ecto-parasitoid Tamarixia radiata (Waterston). Thus, Allium spp. or the tri- and di-sulphides could be integrated into management programmes for D. citri without affecting natural enemies.

Solenopsis invicta Buren (red imported fire ant) are invasive pests that have the capability of major destructive impacts on lifestyle, ecology and economy. Control of this species is dependent, in part, upon ability to estimate the potential spread from newly discovered nests. The potential for spread and the spread characteristics differ between monogyne and polygyne social forms. Prior to this study, differentiation of the two social forms in laboratory test samples commonly used a method involving restriction endonuclease digestion of an amplified Gp-9 fragment. Success of this assay is limited by the quality of DNA, which in the field-collected insects may be affected by temporary storage in unfavourable conditions. Here, we describe an alternative and highly objective assay based upon a high resolution melt technique following preamplification of a significantly shorter Gp-9 fragment than that required for restriction endonuclease digestion. We demonstrate the application of this assay to a S. invicta incursion in Queensland, Australia, using field samples from which DNA may be partially degraded. The reductions in hands-on requirements and overall duration of the assay underpin its suitability for high-throughput testing.

Entedon erythrinae sp. n. (Hymenoptera: Eulophidae), a gregarious egg-larval endoparasitoid of the Erythrina bruchine Specularius impressithorax, an invasive pest of the coral tree seeds (Erythrina spp.), is described from the Hawaiian Islands and Africa (South Africa, Tanzania and Mozambique). The biology and morphology of preimaginal stages of this new species are described in details.

It is remarkable that the early embryo of the parasitoid represents a mass of undifferentiated cells surrounded by a peculiar membrane formed by the peripheral enlarged polygonal cells. The young larva developing inside this membrane corresponds morphologically to the second instar of congeneric species. Various peculiarities of the parasitoid-host relationships in gregarious and solitary Entedon parasitoids are discussed. The DNA sequences of 28S D2 (nuclear), Cytochrome Oxidase I (COI, mitochondrial) and Cytochrome B (CytB, mitochondrial) genes are provided for this new species and compared with the sequences of some other Afrotropical and Palearctic species of the genus.

The Warehouse beetle, Trogoderma variabile (Coleoptera: Dermestidae), is an internationally significant invasive pest of packed goods and stored grain. When it was first documented in Australia at Griffith, New South Wales, in 1977, an eradication campaign was initiated. After several years and considerable effort, the eradication campaign was abandoned. To monitor the presence and spread of T. variabile, surveys were carried out by government agencies in 1992 and 2002. When survey data was compared, it was concluded that the distribution of morphologically identified T. variabile had doubled in most Australian states. Here, we used samples from the 2002 survey to conduct a phylogenetic study using partial sequences of mitochondrial genes Cytochrome oxidase I and Cytochrome B, and the nuclear gene 18S, to examine the distribution and dispersal of T. variabile and detect the presence of misidentified species. Based on our molecular results, we show that only 47% of the samples analysed were T. variabile, and the remaining were a mixture of six putative species. In addition, T. variabile was found in only 78% of the trapping sites. We discuss the importance of correct diagnosis in relation to the eradication campaign.

The Walnut Husk Fly, Rhagoletis completa Cresson (Diptera: Tephritidae), is native to North America (Midwestern US and north-eastern Mexico) and has invaded several European countries in the past decades by likely crossing the alpine divide separating most parts of Switzerland from Italy. Here, we determined its current distribution in Switzerland by sampling walnuts (Juglans regia L.) in ecologically and climatically distinct regions along potential invasion corridors. R. completa was found to be firmly established in most low altitude areas of Switzerland where walnuts thrive, but notably not a single parasitoid was recovered from any of the samples. Infested fruit was recovered in 42 of the 71 localities that were surveyed, with mean fruit infestation rate varying greatly among sites. The incidence of R. completa in Switzerland is closely related to meteorological mean spring temperature patterns influencing growing season length, but not to winter temperatures, reflecting survival potential during hibernation. Importantly, areas in which the fly is absent correspond with localities where the mean spring temperatures fall below 7°C. Historical data records show that the natural cold barrier around the Alpine divide in the central Swiss Alps corresponding to such minimal temperatures has shrunk significantly from a width of more than 40 km before 1990 to around 20 km after 2000. We hypothesize on possible invasion/expansion routes along alpine valleys, dwell on distribution patterns in relation to climate, and outline future research needs as the incursion of R. completa into Switzerland; and, more recently, other European countries, such as Germany, Austria, France and Slovenia, represent an example of alien species that settle first in the Mediterranean Basin and from there become invasive by crossing the Alps.

In cyclical parthenogens, clonal diversity is expected to decrease due to selection and drift during the asexual phase per number of asexual generations. The decrease in diversity may be counteracted by immigration of new genotypes. We analysed temporal variation in clonal diversity in colonies of the monophagous tansy aphid, Macrosiphoniella tanacetaria (Kaltenbach), sampled four times over the course of a growing season. In a related field study, we recorded aphid colony sizes and the occurrence of winged dispersers throughout the season. The number of colonies increased from April, when asexual stem mothers hatched from the sexually produced eggs, to the end of June. The proportion of colonies with winged individuals also increased over this period. After a severe reduction in colony sizes in late summer, a second expansion phase occurred in October when sexuals were produced. At the season's end, the only winged forms were males. A linked genetic study showed that the number of microsatellite multilocus genotypes and genetic variability assessed at three polymorphic loci per colony decreased from June to October. Overall, the relatedness of wingless to winged individuals within colonies was lower than average relatedness among wingless individuals, suggesting that winged forms mainly originated in different colonies. The results demonstrate that patterns of genetic diversity within colonies can be explained by the antagonistic forces of clonal selection, migration and genetic drift (largely due to midsummer population bottlenecks). We further suggest that the males emigrate over comparatively longer distances than winged asexual females.