Esterase-based resistance in the peach-potato aphid, Myzus persicae (Sulzer), is generally due to one of two alternative amplified carboxylesterase genes, E4 or FE4 (fast E4). The E4 amplified form is distributed worldwide and it is correlated with a particular translocation between autosomes 1 and 3, whereas the FE4 form, which has hitherto not been found to be associated with chromosomal rearrangements, is typical of the Mediterranean regions. In this study, we present for the first time cytogenetic and molecular data on some M. persicae parthenogenetic lineages, which clearly show a chromosomal A1-3 translocation associated with esterase FE4 genes and unrelated to high levels of esterase-based resistance.

During the last decade, multilocus analysis has gradually become a powerful tool for the studies of population genetics and phylogeography. The double-striped cockroach, Blattella bisignata, is endemic to southeast Asia, and there is currently little genetic information available for the species. We chose it as the target species to investigate a biodiversity hotspot in southwest China. Here, we report the identification and characterization of 11 single-copy anonymous nuclear markers with an average length of 378bp. These loci, isolated from a genomic library of B. bisignata, can amplify in two additional Blattella species (B. germanica and B. lituricollis). While testing these markers in representative species of Blattellidae, Blattidae and Epilampridae, some of them can cross-amplify successfully. After sequencing 30 individuals collected from southern China per locus, we found relatively high variability (approximately 3.6 SNPs per 100bp). Finally, a small-scale study was also performed to show that these markers do indeed fulfill the expectations as phylogeographic markers.

To estimate the net effect of climate change on natural populations, we must take into account the positive and negative effects of temperature oscillations and climate variability. Warming because of climate change will likely exceed the physiological optima of tropical insects, which currently live very close to their thermal optima. Tropical insects will be negatively affected if their optima are exceeded otherwise warming may affect them positively. We evaluate the demographic responses of the cowpea aphid, Aphis craccivora, to summer warming in subtropical and tropical Taiwan, and examine the effects of diel temperature oscillation on these responses. Aphids were reared at four temperatures (current summer mean, +1.4, +3.9 and +6.4 °C), the latter three simulating different levels of warming. At each average temperature, aphids experienced constant or oscillating (from −2.9 to +3.6 °C of each mean temperature) regimes. As the simulated summer temperatures increased, so did the negative effects on life-history traits and demographic parameters. Compared with aphids reared in constant temperatures, aphids reared in oscillating temperatures developed more slowly and had a longer mean generation time, but their net reproductive rate was higher. These findings demonstrate that climate warming will affect demographic parameters and life-history traits differentially. Studies that use constant temperatures are unlikely to accurately predict biotic responses to climate change.

Interpreting spatial patterns in the abundance of species over time is a fundamental cornerstone of ecological research. For many species, this type of analysis is hampered by datasets that contain a large proportion of zeros, and data that are overdispersed and spatially autocorrelated. This is particularly true for insects, for which abundance data can fluctuate from zero to many thousands in the space of weeks. Increasingly, an understanding of the ways in which environmental variation drives spatial and temporal patterns in the distribution, abundance and phenology of insects is required for management of pests and vector-borne diseases. In this study, we combine the use of smoothing techniques and generalised linear mixed models to relate environmental drivers to key phenological patterns of two species of biting midges, Culicoides pulicaris and C. impunctatus, of which C. pulicaris has been implicated in transmission of bluetongue in Europe. In so doing, we demonstrate analytical tools for linking the phenology of species with key environmental drivers, despite using a relatively small dataset containing overdispersed and zero-inflated data. We demonstrate the importance of landcover and climatic variables in determining the seasonal abundance of these two vector species, and highlight the need for more empirical data on the effects of temperature and precipitation on the life history traits of palearctic Culicoides spp. in Europe.

The Colorado potato beetle (Leptinotarsa decemlineata, family Chrysomelidae), a phytophagous insect, which feeds preferably on potatoes, constitutes a serious pest of this crop and causes extensive damage to tomatoes and eggplants. It has a remarkable ability to develop resistance quickly against insecticides and shows a diversified and flexible life history. Consequently, the control of this pest has become difficult, requiring the development of new alternative biotechnology-based strategies. Such strategies require a thorough knowledge of the beetle's genome, including the repetitive DNA. Satellite DNA (stDNA), composed of long arrays of tandemly arranged repeat units, constitutes the major component of heterochromatin and is located mainly in centromeric and telomeric chromosomal regions. We have studied two different unrelated satellite-DNA families of which the consensus sequences were 295 and 109 bp in length, named LEDE-I and LEDE-II, respectively. Both were AT-rich (70.8% and 71.6%, respectively). Predictive models of sequence-dependent DNA bending and the study of electrophoretic mobility on non-denaturing polyacrylamide gels have shown that the DNA was curved in both satellite-DNA families. Among other features, the chromosome localization of both stDNAs has been studied. In situ hybridization performed on meiotic and mitotic nuclei showed chromosomes, including the X chromosome, with zero, one, or two stDNAs. In recent years, it has been proposed that the repetitive DNA may play a key role in biological diversification processes. This is the first molecular and cytogenetic study conducted on L. decemlineata repetitive DNA and specifically on stDNA, which is one of the important constituents of eukaryotic genomes.

In insects, the enzyme trehalase plays a crucial role in energy metabolism, chitin synthesis and possibly during plant–insect interactions. We have characterized a soluble trehalase gene (Tre-1) from cDNA of Aphis glycines, a serious migratory pest of soybean. The full-length cDNA of Tre-1 in A. glycines (AyTre-1) was 2550 bp long with an open reading frame of 1770 bp that encoded for a 589 amino acid residues protein. Sequence assessment and phylogenetic analysis of the putative protein suggested that the selected cDNA belongs to soluble trehalase group. Quantitative PCR (qPCR) analysis in different tissues and developmental stages revealed peak mRNA levels of AyTre-1 in the gut (compared with other tissues assayed) and highest expression in the second instar compared with the other developmental stages assayed. Interestingly, a significantly increased expression of AyTre-1 (1.9-fold, P < 0.05) was observed in the alate morphs compared with that in apterate morphs. However, there was no significant difference in AyTre-1 expression in A. glycines-nymphs fed with resistant and susceptible plants. Expression patterns identified in this study provide a platform to investigate the role of AyTre-1 in physiological activities such as flight and feeding in A. glycines. The characterization of soluble trehalase gene may help to develop novel strategies to manage A. glycines using trehalase inhibitors and using RNA interference for knock-down of AyTre-1 expression.

Lycorma delicatula (White) was identified in 2004 as an invasive pest in South Korea, where it causes serious damage to vineyard crops. To investigate the population structure and dispersal pattern of L. delicatula in South Korea, we estimated the population genetic structure and gene flow among nine locations across the country using seven microsatellite markers. Although L. delicatula spread throughout most of its geographical range in South Korea within 5–7 years following invasion, its populations show evidence of genetic structuring across the range with a low but significant global FST (genetic differentiation across all populations) of 0.0474. Bayesian-based clustering analysis indicates the presence of at least three genetically unique populations in South Korea, including populations in northeastern South Korea, which show a distinct genetic background. However, isolation by distance suggests that populations in South Korea have not yet reached genetic equilibrium. Estimates of the historical rate of gene flow (Nem) indicate that relatively high rates of flow have been maintained among populations within the western region, which may indicate recent range expansion. A population assignment test using the first-generation migrant detection method suggested that long-distance dispersal of L. delicatula may have occurred over large areas of South Korea. More complex dispersal patterns may have occurred during L. delicatula invasion of heterogeneous landscapes in South Korea.

Transgenic (Bt) cotton dominates Australian cotton production systems. It is grown to control feeding damage by lepidopteran pests such as Helicoverpa armigera. The possibility that these moths might become resistant to Bt remains a threat. Consequently, refuge crops (with no Bt) must be grown with Bt cotton to produce large numbers of Bt-susceptible moths to reduce the risk of resistance developing. A key assumption of the refuge strategy, that moths from different host plant origins mate at random, remains untested. During the period of the study reported here, refuge crops included pigeon pea, conventional cotton (C3 plants), sorghum or maize (C4 plants). To identify the relative contributions made by these (and perhaps other) C3 and C4 plants to populations of H. armigera in cotton landscapes, we measured stable carbon isotopes (δ13C) within individual moths captured in the field. Overall, 53% of the moths were of C4 origin. In addition, we demonstrated, by comparing the stable isotope signatures of mating pairs of moths, that mating is indeed random amongst moths of different plant origins (i.e. C3 and C4). Stable nitrogen isotope signatures (δ15N) were recorded to further discriminate amongst host plant origins (e.g. legumes from non-legumes), but such measurements proved generally unsuitable. Since 2010, maize and sorghum are no longer used as dedicated refuges in Australia. However, these plants remain very common crops in cotton production regions, so their roles as ‘unstructured’ refuges seem likely to be significant.

Acetylcholinesterase (AChE), encoded by the ace gene, is a key enzyme of cholinergic neurotransmission. Insensitive acetylcholinesterase (AChE) has been shown to be responsible for resistance to OPs and CBs in a number of arthropod species, including the most important pest of olives trees, the olive fruit fly Bactrocera oleae. In this paper, the organization of the B. oleae ace locus, as well as the structural and functional features of the enzyme, are determined. The organization of the gene was deduced by comparison to the ace cDNA sequence of B. oleae and the organization of the locus in Drosophila melanogaster. A similar structure between insect ace gene has been found, with conserved exon-intron positions and junction sequences. The B. oleae ace locus extends for at least 75 kb, consists of ten exons with nine introns and is mapped to division 34 of the chromosome arm IIL. Moreover, according to bioinformatic analysis, the Bo AChE exhibits all the common features of the insect AChE. Such structural and functional similarity among closely related AChE enzymes may implicate similarities in insecticide resistance mechanisms.

Host-seeking female tabanid flies, that need mammalian blood for the development of their eggs, can be captured by the classic canopy trap with an elevated shiny black sphere as a luring visual target. The design of more efficient tabanid traps is important for stock-breeders to control tabanids, since these blood-sucking insects can cause severe problems for livestock, especially for horse- and cattle-keepers: reduced meat/milk production in cattle farms, horses cannot be ridden, decreased quality of hides due to biting scars. We show here that male and female tabanids can be caught by a novel, weather-proof liquid-filled black tray laid on the ground, because the strongly and horizontally polarized light reflected from the black liquid surface attracts water-seeking polarotactic tabanids. We performed field experiments to reveal the ideal elevation of the liquid trap and to compare the tabanid-capturing efficiency of three different traps: (1) the classic canopy trap, (2) the new polarization liquid trap, and (3) the combination of the two traps. In field tests, we showed that the combined trap captures 2.4–8.2 times more tabanids than the canopy trap alone. The reason for the larger efficiency of the combined trap is that it captures simultaneously the host-seeking female and the water-seeking male and female tabanids. We suggest supplementing the traditional canopy trap with the new liquid trap in order to enhance the tabanid-capturing efficiency.

Two types of municipal solid waste (MSW), newly arrived and 2 weeks old, were sampled from a sanitary landfill in Pulau Pinang, Malaysia at a fortnightly interval and kept under field conditions for 2 weeks. A total of 480 kg of each type of MSW was sampled to study species composition and impact of delays in cover soil applications on filth fly emergence. Out of 960 kg of MSW sampled, 9.2 ± 0.5 flies emerged per kilogram. Weekly adult fly emergence rates of newly arrived and 2-week-old waste did not differ significantly and MSW remained suitable for fly breeding for up to 1 month. Eight species of flies emerged from the MSW: namely, Musca domestica, Musca sorbens, Synthesiomyia nudiseta, Hydrotaea chalcogaster, Chrysomya megacephala, Lucilia cuprina, Hemipyrellia ligurriens and Sarcophaga sp. Newly arrived waste was determined to be the main source for M. domestica, C. megacephala and L. cuprina in the landfill owing to significantly higher mean emergence compared with 2-week-old waste. Musca sorbens was found in newly arrived waste but not in 2-week-old waste, suggesting that the species was able to survive transportation to landfill but unable to survive landfill conditions. Hemipyrellia ligurriens, H. chalcogaster and S. nudiseta were not imported into the landfill with MSW and pre-existing flies in and around the landfill itself may be their source. The results show that landfills can be a major source of fly breeding if cover soil or temporary cover is not applied daily or on a regular schedule.

Several whitefly species (Hemiptera: Aleyrodidae) are cosmopolitan phloem-feeders that cause serious damage in numerous agricultural crops. All whitefly species harbor a primary bacterial symbiont and a diverse array of secondary symbionts which may influence several aspects of the insect's biology. We surveyed infections by secondary symbionts in Bemisia tabaci (Gennadius), Trialeurodes vaporariorum (Westwood) and Siphoninus phillyreae (Haliday) from areas in the east cost of the Adriatic Sea. Both the Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) B. tabaci genetic groups were detected in Montenegro, whereas only the MED was confirmed in Croatia. Trialeurodes vaporariorum and S. phillyreae were found in all areas surveyed. MEAM1 and MED exhibited similarity to previously reported infections, while populations of T. vaporariorum from Montenegro harbored Rickettsia, Wolbachia and Cardinium in addition to previously reported Hamiltonella and Arsenopnohus. Siphoninus phillyreae harbored Hamiltonella, Wolbachia, Cardinium and Arsenophonus, with the latter appearing in two alleles. Multiple infections of all symbionts were common in the three insect species tested, with some reaching near fixation. Florescent in situ hybridization showed new localization patterns for Hamiltonella in S. phillyreae, and the morphology of the bacteriosome differed from that observed in other whitefly species. Our results show new infections with bacterial symbionts in the whitefly species studied. Infections with the same symbionts in reproductively isolated whitefly species confirm complex relationships between whiteflies and bacterial symbionts, and suggest possible horizontal transfer of some of these bacteria.

Anthropogenic range expansion and cultural practices have modified the distribution, abundance and genetic diversity of domesticated organisms, thereby altering multitrophic assemblages through space and time. The putative Mesoamerican domestication centre of the common bean, Phaseolus vulgaris L., in Mexico allows investigating the effects of plant domestication on the genetic structure of members of a multitrophic system. The aim of this study was to compare the evolutionary history of Horismenus parasitoids (Hymenoptera: Eulophidae) to those of their bruchid beetle hosts (Coleoptera: Bruchidae) and their domesticated host plant (P. vulgaris), in the context of traditional agriculture in Mexico. We analyzed the population genetic structure of four Horismenus species in Mexico using mitochondrial COI haplotype data. The two most abundant parasitoid species were Horismenus depressus and Horismenus missouriensis. Horismenus missouriensis were infected by Wolbachia endosymbionts and had little to no population differentiation (FST = 0.06). We suspect the mitochondrial history of H. missouriensis to be blurred by Wolbachia, because differentiation among infected vs. non-infected individuals exists (FST = 0.11). Populations of H. depressus were found to be highly differentiated (FST = 0.34), but the genetic structuring could not be explained by tested spatial components. We then compared the genetic structure observed in this parasitoid species to previously published studies on bruchid beetles and their host plants. Despite extensive human-mediated migration and likely population homogenization of its two Acanthoscelides bruchid beetle hosts, H. depressus populations are structured like its host plant, by a recent dispersal from a diverse ancestral gene pool. Distinct evolutionary dynamics may explain inconsistent patterns among trophic levels. Parasitoids likely migrate from wild bean populations and are poorly adapted to bean storage conditions similar to their bruchid beetle hosts. Integrating several trophic levels to the study of evolutionary history has proven to be fruitful in detecting different ecological responses to human-mediated disturbances and host parasite interactions.

Wheat bulb fly (WBF) larvae use chemotaxis to orientate towards host-plant root exudates. This study aimed to investigate the role of the primary plant metabolite carbon dioxide (CO2) in host-plant location by WBF. Arena based behavioural experiments were used to identify whether CO2 induced chemotaxis (directional movement in response to a chemical stimulus) or kinesis (non-directional movement in response to a stimulus) from WBF larvae. No chemotactic response was observed when larvae were presented to a point source of CO2. However, elevated levels of CO2 induced kinesis, with both track length and tortuosity (number of twists and turns in the movement path) increasing at elevated CO2 levels of 1000–2000 ppm, demonstrating increased searching behaviour. Soil emission of CO2 was quantified to compare soil levels with those identified as eliciting behavioural effects on the larvae. Samples removed from soil gave a mean CO2 concentration of 557 (±46) ppm, which is lower than the lowest concentration of CO2 found to induce a behavioural response and higher than the lowest CO2 concentration tested, which was found not to alter behaviour. It is proposed that increased CO2 concentrations in the soil act as a behavioural trigger, inducing intensive searching of an area by WBF larvae. This increases the likelihood of finding more host-specific identifiers, such as secondary metabolites when near a potential host-plant.

Anopheles stephensi – Liston (Culicidae: Diptera) is an important urban malarial vector in the Indian sub-continent, accounting for about 15% of the total annual malaria incidence. Chemical control represents a key strategy in the management of this insect vector. However, owing to erratic and continuous application of insecticides, resistance has become a common phenomenon among them and their control has become an uphill task. The genetics of alphamethrin, a synthetic pyrethroid resistance was studied to determine its mode of inheritance. The late third instar larvae were selectively inbred for 27 and ten generations to synthesize homozygous resistant (R) and susceptible (S) stocks, respectively, to the diagnostic dose of 0.12 mg l−1. The log-dosage probit mortality relationships and degree of dominance (D) were calculated. Resistance was observed in both sexes, the dosage-mortality (d-m) line of F1 was towards the resistant parent and the ‘D’ value was found to be 0.8 indicating alphamethrin resistant (amr) gene to be autosomal and incompletely dominant. The d-m lines of F2/backcross exhibited a clear plateau of mortality across a range of doses indicating monogenic resistance. The null hypothesis for monogenic resistance was tested from mortality data of backcross progeny compared with theoretical expectations using the χ2 test and was found to be non-significant. Understanding genetics of insecticide resistance is significant in prediction and management of resistant insects. The amr genes can be used as genetic marker in A. stephensi, which can be used in several applications in conducting basic and applied genetic research.

The greenbug, Schizaphis graminum (Rondani), is a major pest of cereals in some parts of the world and is of particular concern because it can be resistant to some insecticides and overcome the resistance of crops. In the UK, it has never been found on crops, but two rather little-known and closely-related species (Schizaphis holci and Schizaphis agrostis) are associated with the wild grasses, Holcus lanatus and Agrostis stolonifera. Since 1987, winged (alate) aphids morphologically resembling the greenbug have been found in increasing numbers in 12.2 m high suction-trap samples of the Rothamsted Insect Survey (RIS); hence, studies were undertaken to establish their identity. Clones (=asexual lineages) established from populations collected from H. lanatus in southern England showed strong preference for Holcus over Agrostis and Hordeum in laboratory tests and produced sexual morphs when transferred to short-day conditions, the males being apterous, as expected for S. holci. Multivariate morphometric comparisons of alatae caught in UK RIS suction traps in 2007 and 2011 with named specimens from museum collections, including S. graminum from many countries, indicated that the suction-trapped alatae were mostly S. agrostis and S. holci. Cytochrome c oxidase subunit I (COI) mtDNA obtained from 62 UK specimens from suction-traps had 95.4–100% sequence identity with US specimens of S. graminum. Two of the UK specimens had identical COI sequence to the US sorghum-adapted form of S. graminum, and these specimens also had 100% identity with a 640 bp fragment of nDNA CytC, indicating that this form of S. graminum may already be present in the UK. Present and future economic implications of these results are discussed.

Trissolcus basalis has been used as a biological control agent of its main host, Nezara viridula, in many countries. However, estimations of its functional and numerical responses in the field are lacking. We estimated the density of N. viridula eggs, the proportion of parasitized N. viridula eggs, and the number of T. basalis adults/trap in the field. We transformed relative parasitoid density to an absolute density, and estimated the parasitoid's attack rate, a, and the mutual interference parameter, m, in two ways: following Arditi & Akçakaya (1990) and using the Holling–Hassell–Varley model with two iterative techniques. The attack rate estimated by both methods were a=1.097 and a=0.767, respectively. Parameter m varied less between methods: m=0.563 and m=0.586, respectively, and when used to calculate the number of parasitized N. viridula eggs per m2, differences with the observed values were not significant. The numerical response of T. basalis was affected by the sex allocation of their progeny and the proportion of adult parasitoids trapped decreased with field parasitoid population density. Theoretical models show that 0<m<1 is a stabilizing factor and previous re-analysis of field data showed a mean m value of 0.8. The Holling–Hassell–Varley model leads to a flexible description of the functional response allowing to predict acceptable weekly host parasitism. The pre-imaginal parasitoid survival and the change in sex ratio as a function of parasitoid density adequately describe the numerical functional response of the parasitoid.

Probing behaviour (prior to parturition) and parturition of two clones (PS01 and N116) of the pea aphid, Acyrthosiphon pisum on two genotypes (near-isogenic lines (NILs)) (Q174_5.13 and Q174_9.10) of Medicago truncatula were investigated using electrical penetration graph (EPG) coupled with simultaneous visual monitoring for parturition. Line Q174_5.13 has been reported to show a phloem-based resistance to feeding in the clone PS01 but to be susceptible to the clone N116, whereas Q174_9.10 has shown to be susceptible to both aphid clones. The time taken to first parturition by clone PS01 was similar on Q174_5.13 and Q174_9.10. Prior to parturition, no aphids on Q174_5.13 contacted phloem, but 5% of the aphids on Q174_9.10 showed phloem salivation (recognized by EPG pattern E1). No phloem contact was observed with aphid clone N116 on either NILs of Medicago before first parturition occurred, and the time taken to first larviposition was similar on Q174_5.13 and Q174_9.10. The results indicate that the initiation of parturition of the clone PS01 and N116 on both NILs does not require the phloem contact and seems unchanged by a phloem-based resistance mechanism to feeding on Medicago. This finding suggests that host recognition and decisions about parturition occur before phloem contact or ingestion, and act independently on R-gene-mediated resistance.

Ribonucleic acid interference (RNAi) is a sequence-specific gene silencing mechanism induced by double-stranded RNA (dsRNA). Recently, RNAi has gained popularity as a reverse genetics tool owing to its tremendous potential in insect pest management, which includes Helicoverpa armigera. However, its efficiency is mainly governed by dsRNA concentration, frequency of application, target gene, etc. Therefore, to obtain a robust RNAi response in H. armigera, we evaluated various concentrations of dsRNA and its frequency of applications delivered through diet in silencing a midgut gene, chymotrypsin and a non-midgut gene, juvenile hormone acid methyl transferase (jhamt) of H. armigera. The extent of target gene silencing was determined by employing reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Our study revealed four significant findings: (i) single application of dsRNA elicited a delayed and transient silencing, while multiple applications resulted in early and persistent silencing of the above genes; (ii) silencing of the non-midgut gene (jhamt) through diet delivered dsRNA revealed prevalence of systemic silencing probably due to communication of silencing signals in this pest; (iii) the extent of silencing of chymotrypsin was positively correlated with dsRNA concentration and was negatively correlated with jhamt; (iv) interestingly, over-expression (15–18 folds) of an upstream gene, farnesyl diphosphate synthase (fpps), in juvenile hormone (JH) biosynthetic pathway at higher concentrations of jhamt dsRNA was the plausible reason for lesser silencing of jhamt. This study provides an insight into RNAi response of target genes, which is essential for RNAi design and implementation as a pest management strategy.

To advance our restricted knowledge on mosquito biodiversity and distribution in Belgium, a national inventory started in 2007 (MODIRISK) based on a random selection of 936 collection points in three main environmental types: urban, rural and natural areas. Additionally, 64 sites were selected because of the risk of importing a vector or pathogen in these sites. Each site was sampled once between May and October 2007 and once in 2008 using Mosquito Magnet Liberty Plus traps. Diversity in pre-defined habitat types was calculated using three indices. The association between species and environmental types was assessed using a correspondence analysis. Twenty-three mosquito species belonging to traditionally recognized genera were found, including 21 indigenous and two exotic species. Highest species diversity (Simpson 0.765) and species richness (20 species) was observed in natural areas, although urban sites scored also well (Simpson 0.476, 16 species). Four clusters could be distinguished based on the correspondence analysis. The first one is related to human modified landscapes (such as urban, rural and industrial sites). A second is composed of species not associated with a specific habitat type, including the now widely distributed Anopheles plumbeus. A third group includes species commonly found in restored natural or bird migration areas, and a fourth cluster is composed of forest species. Outcomes of this study demonstrate the effectiveness of the designed sampling scheme and support the choice of the trap type. Obtained results of this first country-wide inventory of the Culicidae in Belgium may serve as a basis for risk assessment of emerging mosquito-borne diseases.