Sexual maturation of Anastrepha fraterculus is a long process. Methoprene (a mimic of juvenile hormone) considerably reduces the time for sexual maturation in males. However, in other Anastrepha species, this effect depends on protein intake at the adult stage. Here, we evaluated the mating competitiveness of sterile laboratory males and females that were treated with methoprene (either the pupal or adult stage) and were kept under different regimes of adult food, which varied in the protein source and the sugar:protein ratio. Experiments were carried out under semi-natural conditions, where laboratory flies competed over copulations with sexually mature wild flies. Sterile, methoprene-treated males that reached sexual maturity earlier (six days old), displayed the same lekking behaviour, attractiveness to females and mating competitiveness as mature wild males. This effect depended on protein intake. Diets containing sugar and hydrolyzed yeast allowed sterile males to compete with wild males (even at a low concentration of protein), while brewer´s yeast failed to do so even at a higher concentration. Sugar only fed males were unable to achieve significant numbers of copulations. Methoprene did not increase the readiness to mate of six-day-old sterile females. Long pre-copulatory periods create an additional cost to the management of fruit fly pests through the sterile insect technique (SIT). Our findings suggest that methoprene treatment will increase SIT effectiveness against A. fraterculus when coupled with a diet fortified with protein. Additionally, methoprene acts as a physiological sexing method, allowing the release of mature males and immature females and hence increasing SIT efficiency.

Species distribution models (SDMs), which are well established in many fields of biological research, are still uncommon in the agricultural risk analysis of pest insects. To exemplify the use of SDMs, we investigated the influence of environmental factors on the occurrence of Hyalesthes obsoletus Signoret (Hemiptera: Cixiidae). The planthopper is the only known vector of the grapevine yellows disease ‘bois noir’. The study was conducted in 145 locations in the Baden region of southwest Germany. The planthopper was surveyed on host plant patches, consisting of stinging nettle and/or bindweeds. We used a stratified modelling framework where (1) species presence–absence data were related to an extensive environmental dataset using logistic regressions; and (2) different types of average models were developed based on an information theoretic method. The results show that the incidence of H. obsoletus is associated to above- as well as below-ground environmental factors, particularly to the amount of fine soil and average annual precipitation. This result was consistent across all average models. The relative importance of other environmental variables was dependent upon the average model under consideration and thus may vary according to their intended use, either the explanation of habitat requirements or the prediction and mapping of occurrence risks. The study showed that SDMs offer a quantification of species’ habitat requirements and thus, could represent a valuable tool for pest management purposes. By providing examples of current issues of grapevine pests in viticulture, we discuss the use of SDMs in agricultural risk analysis and highlight their advantages and caveats.

Some bark beetle species (Coleoptera: Scolytinae) produce aggregation pheromones that allow coordinated attack on their conifer hosts. As a new saproxylic habitat is founded, an assemblage of associated beetles kairomonally respond to bark beetle infochemicals. Ips sexdentatus is one of the major damaging insects of Pinus spp. in Southern Europe. Its response to varying ipsenol (Ie) percentages in relation to ipsdienol (Id) was studied in northwestern Spain, along with the entire saproxylic beetle assemblage captured at multiple-funnel traps. Response profile modeling was undertaken for I.sexdentatus sexes and sex-ratios, associated species and for selected trophic groups using a reference Gaussian model. In addition, the effects on the saproxylic assemblages were analyzed. I. sexdentatus response curve peaked at 22.7% Ie content, while remaining taxa that could be modeled, peaked above ca. 40% Ie. Predator guilds showed a linear relationship with Ie proportion, while competitors showed a delayed response peak. Consequently, species assemblages differed markedly between varying pheromone component mixtures. Given that the evaluated pheromonal proportions mimicked that of logs being colonized by I. sexdentatus, results suggested that the registered differential responses at different levels might provide I.sexdentatus with a temporal window that maximizes conspecific attraction while reducing interference with competitor and predatory guilds. Described responses might help improve the monitoring of the population status of target bark beetles and their associates, but also point toward the by-catch of many natural enemies, as well as rare saproxylic beetle species, interfering with the aims of sustainable forest management.

Dry grain legume seeds possessing αAI-1, an α-amylase inhibitor from common bean (Phaseolus vulgaris), under the control of a cotyledon-specific promoter have been shown to be highly resistant to several important bruchid pest species. One transgenic chickpea and four cowpea lines expressing αAI-1, their respective controls, as well as nine conventional chickpea cultivars were assessed for their resistance to the bruchids Acanthoscelides obtectus (Say), Callosobruchus chinensis L. and Callosobruchus maculatus F. All transgenic lines were highly resistant to both Callosobruchus species. A. obtectus, known to be tolerant to αAI-1, was able to develop in all transgenic lines. While the cotyledons of all non-transgenic cultivars were highly susceptible to all bruchids, C. chinensis and C. maculatus larvae suffered from significantly increased mortality rates inside transgenic seeds. The main factor responsible for the partial resistance in the non-transgenic cultivars was deduced to reside in the seed coat. The αAI-1 present in seeds of transgenic chickpea and cowpea lines significantly increases their resistance to two important bruchid pest species (C. chinensis and C. maculatus) essentially to immunity. To control αAI-1 tolerant bruchid species such as A. obtectus and to avoid the development of resistance to αAI-1, varieties carrying this transgene should be protected with additional control measures.

Insect cell lines provide useful in vitro models for studying biological systems, including interactions between mosquitoes and obligate intracellular endosymbionts such as Wolbachia pipientis. The Aedes albopictus Aa23 cell line was the first cell line developed to allow examination of Wolbachia infections. However, Wolbachia studies using Aa23 can be complicated by the presence of different cell types in the cell line and the substantial temporal variation in infection level. Two approaches were examined to ameliorate infection variability. In the first approach, multiple Aa23 passaging regimes were tested for an effect on infection variability. Fluorescence in situ hybridization (FISH) staining was used to characterize Wolbachia infection level over time. The results demonstrate an impact of passaging method on Wolbachia infection level, with some methods resulting in loss of infection. None of the passaging methods succeeded in effectively mitigating infection level variation. In a second approach, the clonal C7-10 A. albopictus cell line was infected with Wolbachia from Aa23 cells and Drosophila simulans (Riverside), resulting in cell lines designated C7-10B and C7-10R, respectively. Characterization via FISH staining showed greater stability and uniformity of Wolbachia infection in C7-10R relative to the infection in C7-10B. Characterization of the Aa23, C7-10B and C7-10R lines is discussed as a tool for the study of Wolbachia-host cell interactions.

The field of ecological immunology currently relies on using a number of immune effectors or markers. These markers are usually used to infer ecological trade-offs (via conflicts in resource allocation), though physiological nature of these markers remains elusive. Here, we review markers frequently used in insect evolutionary ecology research: cuticle darkening, haemocyte density, nodule/capsule formation, phagocytosis and encapsulation/melanization via use of nylon filaments and beads, phenoloxidase activity, nitric oxide production, lysozyme and antimicrobial peptide production. We also provide physiologically based information that may shed light on the probable trade-offs inferred when these markers are used. In addition, we provide a number of methodological suggestions to improve immune marker assessment.

Effective, statistically robust sampling and surveillance strategies form an integral component of large agricultural industries such as the grains industry. Intensive in-storage sampling is essential for pest detection, integrated pest management (IPM), to determine grain quality and to satisfy importing nation's biosecurity concerns, while surveillance over broad geographic regions ensures that biosecurity risks can be excluded, monitored, eradicated or contained within an area. In the grains industry, a number of qualitative and quantitative methodologies for surveillance and in-storage sampling have been considered. Primarily, research has focussed on developing statistical methodologies for in-storage sampling strategies concentrating on detection of pest insects within a grain bulk; however, the need for effective and statistically defensible surveillance strategies has also been recognised. Interestingly, although surveillance and in-storage sampling have typically been considered independently, many techniques and concepts are common between the two fields of research. This review aims to consider the development of statistically based in-storage sampling and surveillance strategies and to identify methods that may be useful for both surveillance and in-storage sampling. We discuss the utility of new quantitative and qualitative approaches, such as Bayesian statistics, fault trees and more traditional probabilistic methods and show how these methods may be used in both surveillance and in-storage sampling systems.

Larvae of aphidophagous ladybirds leave a cue in their tracks that deters oviposition. The influence of relatedness on this behaviour is for the first time explored in this paper. Two-spot ladybird females (Adalia bipunctata L.) under different conditions (young and naive, young and experienced, and old and naive) were exposed either to (i) clean filter paper, (ii) filter paper contaminated with unrelated larval tracks or (iii) filter paper contaminated with related larval tracks from their offspring. Oviposition time was recorded during nine hours. Oviposition was inhibited by larval tracks, and was more so by related than unrelated ones. Experienced females showed the same behaviour but with lower intensity. With old females, response was not significant with any type of larval tracks. This is the first report of the effect of relatedness on a ladybird's response to larval tracks.

In Guatemala prior to control initiatives, the main vectors of Trypanosoma cruzi, the causative agent of Chagas disease, were Rhodnius prolixus and Triatoma dimidiata. This study conducted in 2006 in the department of Chiquimula recorded a high level of T. dimidiata infestation and an absence of R. prolixus in all surveyed communities. In Guatemala, the presence of T. dimidiata as domestic, peridomestic and sylvatic populations results in control difficulties as houses are re-infested from the surrounding environment. Entomological surveys, the current method used to select houses in need of control efforts, are labour intensive and time consuming. A time- and cost-effective way to prioritize houses for evaluation and subsequent treatment is the stratification of houses based on the risk of triatomine infestation. In the present study, 17 anthropogenic risk factors were evaluated for associations with house infestation of T. dimidiata including: wall, floor and roof type. There was an increased likelihood of domestic infestation with T. dimidiata associated with the presence of dirt floors (18/29; OR 8.075, 95% CI 2.13–30.6), uncoated bajareque walls (12/17; OR 4.80, 95% CI 1.35–17.1) and triatomine-like faeces on walls (16/26; OR 3.89, 95% CI 1.19–12.7). These factors could be used to target control of T. dimidiata to communities with an increased risk of being infested.

Wheat bulb fly (WBF, Delia coarctata, Fallén, Diptera: Anthomyiidae) is a pest of commercial importance in wheat, barley and rye, with attacked crops failing to produce full potential yields. Females do not oviposit in association with a host-plant; therefore, prompt location of a suitable host is critical to the survival of the newly hatched larvae. The objective of this study was to conduct choice test bioassays to assess the attraction of WBF larvae to specific chemical constituents of WBF host-plant root exudates, the hydroxamic acids DIMBOA (2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one) and MBOA (6-methoxy-benzoxazolin-2-one). The larval response to four concentrations of each test compound was assessed in arena bioassays. Analysis using a Rayleigh test of uniformity of the final resting positions of larvae in response to these chemicals indicated attraction. These results go some way to explaining the mechanisms by which WBF larvae locate host plants, giving the potential to develop semiochemical based control strategies.

The black currant-lettuce aphid, Nasonovia ribisnigri, is an important pest of cultivated lettuce, Lactuca sativa. Since 1982, the control of this aphid on lettuce is largely based on host plant resistance, conferred by the Nr gene, introgressed from Lactuca virosa. The resistance mechanism remains to be identified. N. ribisnigri populations virulent on the Nr-based resistance in lettuce have emerged in several locations in Europe since 2007. The objective of this study was to investigate the resistance mechanism mediated by the Nr gene in lettuce by detailed studies of aphid feeding behaviour and performance. Both avirulent (Nr:0) and virulent (Nr:1) biotypes of N. ribisnigri were studied on five resistant and two susceptible near isogenic lines (NILs). In addition, survival and colony development were quantified. Nr:0 aphids showed a strong decrease in sieve element ingestion and took longer to accept a sieve element on resistant NILs compared with susceptible NILs, and no aphids survived on the resistant NIL. Nr:1 aphids fed and performed equally well on the resistant and susceptible NILs. The resistance mechanism against Nr:0 aphids encoded by the Nr gene seems to be located in the phloem, although we also observed differences in feeding behaviour during the pathway phase to the phloem. Nr:1 aphids were highly virulent to the resistance conferred by the Nr gene. The consequences of the appearance of Nr:1 aphids for control of N. ribisnigri are discussed.

The brown planthoppers (BPH) Nilaparvata lugens (Stål) and the white-backed planthoppers (WBPH) Sogatella furcifera (Horváth) annually migrate from tropical and subtropical regions to temperate regions in Asia, including Japan, Korea and northern China. To elucidate the genetic divergence based on geography of planthoppers and to estimate their migration route on the basis of molecular data, we analysed a part of their mitochondrial genome sequences. Sequences of cytochrome oxidase subunit I (cox1) – transfer RNA for Leu (trnL2) – cox2 were determined for 579 BPH (1,928 bp) and 464 WBPH (1,927 bp) individuals collected from 31 and 25 locations, respectively, in East and Southeast Asia. Thirty and 20 mitochondrial haplotypes were detected for BPH and WBPH, respectively. Single populations of both planthoppers included multiple haplotypes, and many haplotypes were shared in some populations and areas. The most frequently detected haplotypes accounted for approximately 50% of all BPH and WBPH individuals. To evaluate gene flow among planthoppers in different regions in Asia, pairwise fixation index (Fst) values were calculated. For BPH, high Fst values (0.580–0.926) were shown between planthoppers in Papua New Guinea (PNG) and the other areas and moderate Fst values (0.176–0.362) were observed between those in southern Philippines and other areas. For WBPH, the Fst value was the highest between Taiwan and southern Vietnam (0.236), and low among the other areas. AMOVA indicated no genetic structure among eight areas, excluding southern Philippines and PNG, for BPH, and among ten areas for WBPH. These data indicate that both planthoppers do not show much differentiation of local populations and/or have genetically intermixed Asian populations. These data also indicate that it may be difficult to distinguish regional planthopper populations on the basis of differences in mitochondrial sequences.

Plant sugars are often considered as primary feeding stimuli, conditioning host plant acceptance by herbivorous insects. Of the nine sugars identified from methanolic extracts of seven grass species, only turanose, a sucrose isomer, was negatively correlated with the survival and growth of the noctuid larva of cereal stemborer, Busseola fusca. Sucrose was the most abundant sugar, although it did not vary significantly in concentration among the plant species studied. Using Styrofoam™ cylinders impregnated with increasing concentrations of turanose or sucrose, the two sugars had opposing effects: turanose appeared phagodeterrent while sucrose was phagostimulatory. Electrophysiological studies indicated that B. fusca larvae were able to detect both sugars via their styloconic sensilla located on the mouthparts. The findings indicate that, whereas sucrose is a feeding stimulant and positively influences food choice by B. fusca larvae, turanose negatively contributes to larval food choice. The balance in concentrations of both sugars, however, somehow influences the overall host plant choice made by the larvae. This can partly explain host plant suitability and choice by this caterpillar pest in the field.

The synchrony between emergence of spring-active, insect herbivores and the budburst of their host plants could be affected by warming temperatures with influences on the availability and quality of foliage as it undergoes physical and chemical changes. This can affect the growth and survival of insects. Here, we used sun-exposed and shaded trees to determine whether the synchrony between egg hatch of western tent caterpillar, Malacosoma californicum pluviale Dyar (Lepidoptera: Lasiocampidae) and budburst of its host red alder, Alnus rubra Bongard (Betulaceae) changes with different thermal environments (temperature and light together). To explore the potential outcome of a shift in phenological synchrony, we used laboratory assays of larval growth and survival to determine the effect of variation in young, youthful and mature leaves from sun-exposed and shaded trees. While the average higher temperature of sun-exposed trees advanced the timing of budburst and egg hatch, synchrony was not disrupted. Leaf quality had no significant influence on growth or survival in the laboratory for early instars reared as family groups. Later instar larvae, however, performed best on mature leaves from sun-exposed trees. The robust relationship between leaf and larval development of western tent caterpillars suggests that warming climates may not have a strong negative impact on their success through shifts in phenological synchrony, but might influence other aspects of leaf quality and larval condition.

Two stable infections of Wolbachia pipientis, wMelPop and wMel, now established in Aedes aegypti, are being used in a biocontrol program to suppress the transmission of dengue. Any effects of Wolbachia infection on insecticide resistance of mosquitoes may undermine the success of this program. Bioassays of Ae. aegypti were conducted to test for differences in response to insecticides between Wolbachia infected (wMelPop, wMel) and uninfected lines. Insecticides screened were bifenthrin, the pyrethroid commonly used for adult knockdown, as well as larvicides: Bacillus thuringiensis var. israelensis, the organophosphate, temephos and the insect growth regulator, s-methoprene. While differences in response between lines were detected for some insecticides, no obvious or consistent effects related to presence of Wolbachia infection were observed. Spreading Wolbachia infections are, therefore, unlikely to affect the efficacy of traditional chemical control of mosquito outbreaks.

In insect, fat body plays major roles in insect innate immunity. Phenoloxidase (PO) is an important component in insect innate immunity and is necessary for acclimatization. In our study, two prophenoloxidase (PPO) subunits were obtained from fat body of Catantops pinguis (Stål). The full-length cDNA sequence of one PPO (CpPPO1) consisted of 2347 bp with an open reading frame (ORF) of 2187 bp encoding 728 amino acids, while the other subunit (CpPPO2) had a full length of 2445 bp, encoding 691 amino acids. Both the PPO gene products are predicted to possess all the structural features of other PPO members, including two putative tyrosinase copper-binding motifs with six highly conserved histidine residues and a thiolester-like motif. Tissue distribution analysis showed that both PPO mRNAs were abundantly expressed in the fat body among 11 tissues examined, and they were transiently up-regulated after Escherichia coli infection, consistent with them being immune-responsive genes. Total levels of CpPPO1 and CpPPO2 mRNA transcripts were much higher in first instar larvae and adults. A much higher transcript level of CpPPO1 was detected in several months, while there were extremely high mRNA expression levels of CpPPO2 in January, July, October, and December. The above results suggested that PPO from fat body might also bring significant function during the processes of development and acclimatization for C. pinguis.

Ceratovacuna and Pseudoregma are important groups in Cerataphidini (Hemiptera, Hormaphidinae) that not only produce soldier aphids in galls on the primary hosts but also produce horned soldiers on the herbaceous secondary hosts. However, due to sampling bias in previous studies, the phylogenetic relationships of these two genera remain inconclusive. In this study, based on more extensive sampling and examination of both mitochondrial (cytochrome c oxidase subunit I (COI); cytochrome b (Cytb)) and nuclear (elongation factor-1α (EF-1α); long-wavelength opsin (LWO)) genes, we reconstructed the phylogenetic relationships of Ceratovacuna and Pseudoregma. Phylogenetic analyses, along with morphological evidence, suggested that these two genera belong to the paraphyletic groups with species clustered into three main groups. The monophyly of Ceratovacuna and Pseudoregma as a whole was generally supported by all analyses. Monophyly of Pseudoregma was also supported. The estimated divergence times demonstrated that diversification of Ceratovacuna and Pseudoregma occurred approximately at 10 mya. The relatively low resolution for the basal relationships of the three main clades may indicate that these two genera have experienced a rapid radiation along with speciation burst of their secondary hosts during the late Tertiary.

Aphids are major pests in apple orchards, debilitating the crop and spreading disease. We investigated whether early-season predation by canopy spiders may be effectively controlling aphid numbers in three organic orchards. For this purpose, we monitored the aphid population dynamics from the winter eggs to colony stages and compared this to spider abundances and rates of predation on aphids detected by diagnostic polymerase chain reaction. For the latter, we applied existing general aphid primers. We found that spiders ate colony fundatrices and that aphid numbers were negatively related to spider abundance. Spiders were the main active predators within the orchards when the first colony fundatrices were present, indicating their importance in the early control of aphid populations.

Parasitoids of herbivorous insects have frequently evolved specialized lineages exploiting hosts occurring on different plants. This study investigated whether host specialization is also observed when closely related parasitoids exploit herbivorous hosts sharing the same host plant. The question was addressed in economically relevant aphid parasitoids of the Lysiphlebus fabarum group. They exploit two aphid species (Aphis fabae cirsiiacanthoides and Brachycaudus cardui), co-occurring in mixed colonies (syntopy) on the spear thistle (Cirsium vulgare). Two morphologically distinguishable parasitoid lineages of the genus Lysiphlebus were observed and each showed virtually perfect host specialization on one of the two aphid species in this system. From A. f. cirsiiacanthoides, only females emerged that morphologically belonged to Lysiphlebus cardui, while males and females belonging to L. fabarum hatched from B. cardui. Microsatellite analyses indicated clear genetic differentiation of L. fabarum and L. cardui. L. cardui comprised only two distinct asexual lineages, one of which predominated throughout the area investigated. Population genetic analysis of sexual L. fabarum showed evidence for relatively strong spatial structuring and limited dispersal ability. Hyperparasitoids emerged from a large proportion of aphid mummies. One species, Pachyneuron aphidis, was significantly associated with B. cardui/L. fabarum mummies, indicating that host specialization may even extend to the trophic level above parasitoids.

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.