Wild habitats adjoining farmland are potentially valuable sources of natural enemies, but also of pests. Here we tested the utility of birds as ‘sampling devices’, to identify the diversity of prey available to predators and particularly to screen for pests and natural enemies using natural ecosystems as refugia. Here we used PCR to amplify prey DNA from three sympatric songbirds foraging on small invertebrates in Phragmites reedbed ecosystems, namely the Reed Warbler (Acrocephalus scirpaceus), Sedge Warbler (Acrocephalus schoenobaenus) and Cetti's Warbler (Cettia cetti). A recently described general invertebrate primer pair was used for the first time to analyse diets. Amplicons were cloned and sequenced, then identified by reference to the Barcoding of Life Database and to our own sequences obtained from fresh invertebrates. Forty-five distinct prey DNA sequences were obtained from 11 faecal samples, of which 39 could be identified to species or genus. Targeting three warbler species ensured that species-specific differences in prey choice broadened the range of prey taken. Amongst the prey found in reedbeds were major pests (including the tomato moth Lacanobia oleracea) as well as many potentially valuable natural enemies including aphidophagous hoverflies and braconid wasps. Given the mobility of birds, this approach provides a practical way of sampling a whole habitat at once, providing growers with information on possible invasion by locally resident pests and the colonization potential of natural enemies from local natural habitats.

This is the first report of microsatellite markers (simple sequence repeats, SSR) for fall webworm, Hyphantria cunea (Drury) (Lepidoptera: Arctiidae), an important quarantine pest in some European and Asian countries. Here, we developed 48 microsatellite markers for H. cunea from SSR enrichment libraries. Sequences isolated from libraries were sorted into four categories and analyzed. Our results suggest that sequences classified as Grouped should not be used for microsatellite primer design. The genetic diversity of microsatellite loci was assessed in 72 individuals from three populations. The number of alleles per locus ranged from 2 to 5 with an average of 3. The observed and expected heterozygosities of loci ranged from 0 to 0.958 and 0 to 0.773, respectively. A total of 18 out of 153 locus/population combinations deviated significantly from Hardy–Weinberg equilibrium. Moreover, significant linkage disequilibrium was detected in one pair of loci (1275 pairs in total). In the neutral test, two loci were grouped into the candidate category for positive selection and the remainder into the neutral category. In addition, a complex mutation pattern was observed for these loci, and FST performed better than did RST for the estimation of population differentiation in different mutation patterns. The results of the present study can be used for population genetic studies of H. cunea.

The presence of the yellow-necked drywood termite, Kalotermes flavicollis Fabr., has been reported along most of the Mediterranean coasts of Europe, Africa and Asia. While morphological and genetic data exist on European K. flavicollis populations from western and central Mediterranean regions, data on eastern Mediterranean Kalotermes are scarce, and no genetic data exist to date. In this study, we analyzed 17 Kalotermes sp. colonies from 11 localities in the Levant (Cyprus, Lebanon, and Israel), in order to characterize genetically (mitochondrial DNA: COII, 16S, and control region) these populations. We found that samples from the Levant are genetically different from K. flavicollis, with distance values falling in the range of interspecific distances. In the phylogeny of European Kalotermes populations, samples from the Levant form a clade of their own, sister to a clade including K. flavicollis and Kalotermes italicus. Inside the eastern Mediterranean clade, all the samples from Cyprus are included in a well-supported subclade, suggesting that the colonization of the island might have occurred in a single event. These findings show that the populations we examined do not belong to the species K. flavicollis, but to a new species peculiar to the Levant, that we describe as Kalotermes phoeniciae sp. nov. It is possible that previous reports of K. flavicollis in this region can be attributed to K. phoeniciae.

Mortality estimates are central to understanding tsetse fly population dynamics, but are difficult to acquire from wild populations. They can be obtained from age distribution data but, with limited data, it is unclear whether the assumptions required to make the estimates are satisfied and, if not, how violations affect the estimates. We evaluate the assumptions required for existing mortality estimation techniques using long-term longitudinal ovarian dissection data from 144,106 female tsetse, Glossina pallidipes Austen, captured in Zimbabwe between 1988 and 1999. At the end of the hot-dry season each year, mean ovarian ages peaked, and maximum-likelihood mortality estimates declined to low levels, contrary to mark-recapture estimates, suggesting violations of the assumptions underlying the estimation technique. We demonstrate that age distributions are seldom stable for G. pallidipes at our study site, and hypothesize that this is a consequence of a disproportionate increase in the mortality of pupae and young adults at the hottest times of the year. Assumptions of age-independent mortality and capture probability are also violated, the latter bias varying with capture method and with pregnancy and nutritional status. As a consequence, mortality estimates obtained from ovarian dissection data are unreliable. To overcome these problems we suggest simulating female tsetse populations, using dynamical modelling techniques that make no assumptions about the stability of the age distribution.

Wolbachia are intracellular, maternally transmitted bacteria considered the most abundant endosymbionts found in arthropods. They reproductively manipulate their host in order to increase their chances of being transmitted to the offspring, and currently are being used as a tool to control vector-borne diseases. Studies on distribution of Wolbachia among its arthropod hosts are important both for better understanding why this bacterium is so common, as well as for its potential use as a biological control agent. Here, we studied the incidence of Wolbachia in a broad range of insect species, collected from different regions of Brazil, using three genetic markers (16S rRNA, wsp and ftsZ), which varied in terms of their sensitivity to detect this bacterium. The overall incidence of Wolbachia among species belonging to 58 families and 14 orders was 61.9%. The most common positive insect orders were Coleoptera, Diptera, Hemiptera and Hymenoptera, with Diptera and Hemiptera having the highest numbers of Wolbachia-positive families. They included potential human disease vectors whose infection status has never been reported before. Our study further shows the importance of using quantitative polymerase chain reaction for high-throughput and sensitive Wolbachia screening.

The codling moth Cydia pomonella (L.) is an economically important fruit pest and it has been directly targeted by insecticides worldwide. Serious resistance to insecticides has been reported in many countries. As one of the most serious invasive pest, the codling moth has populated several areas in China. However, resistance to insecticides has not been reported in China. We investigated the insecticide-resistance status of four field populations from Northwestern China by applying bioassays, enzyme activities, and mutation detections. Diagnostic concentrations of lambda-cyhalothrin, chlorpyrifos-ethyl, carbaryl, and imidacloprid were determined and used in bioassays. Field populations were less susceptible to chlorpyrifos-ethyl and carbaryl than laboratory strain. Insensitive populations displayed an elevated glutathione S-transferases (GSTs) activity. Reduced carboxylesterase (CarE) activity was observed in some insecticide insensitive populations and reduced acetylcholinesterase activity was observed only in the Wuw population. The cytochrome P450 polysubstrate monooxygenases activities in four field populations were not found to be different from susceptible strains. Neither the known-resistance mutation F399V in the acetylcholinesterase (AChE) gene, ace1, nor mutations in CarE gene CpCE-1 were found in adult individuals from our field populations. Native-PAGE revealed that various CarE isozymes and AChE insensitivity were occurring among Chinese populations. Our results indicate that codling moth populations from Northwestern China were insensitivity to chlorpyrifos-ethyl and carbaryl. Increased GST activity was responsible for insecticides insensitivity. Decreased CarE activity, as well as the presence of CarE and AChE polymorphisms might also be involved in insecticides insensitivity. New management strategies for managing this pest are discussed.

The Asian corn borer, Ostrinia furnacalis enters diapause as fully grown larvae. Owing to geographical variation in photoperiodic control of diapause, the subtropical strain from Hefei city (HF) enters diapause in response to short daylengths, whereas the tropical strain from Ledong county (LD) exhibits almost no diapause under the same conditions. The two strains were used in crosses to study the inheritance of diapause. The HF strain showed a typical long-day response with a critical daylength of approximately14.97 h at 22 °C, 14.60 h at 25 °C and 13.68 h at 28 °C. The LD strain showed weak photoperiodic responses at 22 and 25 °C; and the F1 progeny also showed a long-day response with significantly shorter critical daylength compared with the HF strain. However, the LD × HF (F × M) crosses had significantly longer critical daylengths than HF × LD crosses, indicating a sexual bias in the inheritance of diapause induction, with the male parent having more influence on the F1 progeny. The critical daylength in a backcross to HF was significantly longer than a backcross to LD. Whether the inheritance of diapause fits an additive hypothesis or not depended on photoperiod, with results from different photoperiods showing additive inheritance or incomplete dominance of either diapause or non-diapause. Unlike diapause induction, the duration of diapause for reciprocal crosses was equally influenced by each parent, suggesting that diapause incidence and maintenance are controlled by separate systems in O. furnacalis.

Grasshopper plagues have seriously disturbed grassland ecosystems in Inner Mongolia, China. The accurate prediction of grasshopper infestations and control of grasshopper plagues have become urgent needs. We sampled 234, 342, 335, and 369 plots in Xianghuangqi County of Xilingol League in 2010, 2011, 2012, and 2013, respectively, and measured the density of the most dominant grasshopper species, Oedaleus decorus asiaticus, and the latitude, longitude, and associated relatively stable habitat factors at each plot. We used Excel–GeogDetector software to explore the effects of individual habitat factors and the two-factor interactions on grasshopper density. We estimated the membership of each grasshopper density rank and determined the weights of each habitat category. These results were used to construct a model system evaluating grasshopper habitat suitability. The results showed that our evaluation system was reliable and the fuzzy evaluation scores of grasshopper habitat suitability were good indicators of potential occurrence of grasshoppers. The effects of the two-factor interactions on grasshopper density were greater than the effects of any individual factors. O. d. asiaticus was most likely to be found at elevations of 1300–1400 m, flat terrain or slopes of 4–6°, typical chestnut soil with 70–80% sand content in the top 5 cm of soil, and medium-coverage grassland. The species preferred temperate bunchgrass steppe dominated by Stipa krylovii and Cleistogenes squarrosa. These findings may be used to improve models to predict grasshopper occurrence and to develop management guidelines to control grasshopper plagues by changing habitats.

The Mediterranean corn borer, Sesamia nonagrioides, occurs sympatrically in the northeast of Spain with other lepidopteran pests such as Ostrinia nubilalis and Mythimna unipuncta. In this study, we evaluated the electrophysiological and behavioural response of mated and unmated females and males of S. nonagrioides to their own complete pheromone blend, to its own four components separately, and to the pheromone components of the sympatric species O. nubilalis and M. unipuncta. Results of the electroantennogram recordings revealed that females of S. nonagrioides can detect their own pheromone blend and its individual components. Moreover, our results show that unmated females and males of S. nonagrioides are more sensitive to the female pheromone, showing higher electrophysiological response than the mated females and males. Electroantennogram recordings showed that males and females can detect the major sexual pheromone component of O. nubilalis (Z)-11-tetradecenyl acetate and the minor component of the pheromone of M. unipuncta (Z)-9-hexadecenyl acetate. When the sex pheromone stimulus was presented in the dual-choice assays, gravid females of S. nonagrioides were attracted to both their own complete pheromone blend and one of their own minor pheromone components, (Z)-11-hexadecenal, but the major sexual pheromone component of O. nubilalis acts as a behavioural antagonist to the females.

The egg parasitoid Trissolcus vassilievi (Mayr) (Hymenoptera: Scelionidae) is a significant natural enemy of the sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae), the most important pest of wheat in Iran. This study examined the developmental time and egg-to-adult survival of two geographically separate populations of T. vassilievi on two corresponding host populations at five constant temperatures ranging from 15.0 to 35.0±1°C. No wasps of either population emerged at 15.0°C and the temperature threshold for development was similar between populations, ranging from 13.1±0.3 to 13.8±0.4°C for males and 12.2±0.1 to 12.6±0.1°C for females, but the thermal constant varied with gender and parasitoid population. Development of wasps from the colder Tabriz location was slower, with thermal constants for males and females of 172.6±3.1 and 204.1±1.2 degree-days, respectively, compared to Varamin wasps with 164.7±3.0 and 195.6±1.3 degree-days, respectively. Based on genetic inheritance patterns, reciprocal crosses between the two populations were expected to result in females with thermal phenotypes intermediate to their parental populations, and males that resembled their mothers. However, female progeny of crosses more closely resembled their maternal population, indicating a maternal effect on thermal phenotype. Furthermore, the magnitude of the maternal effect on the thermal constant was asymmetric and was more strongly expressed by Varmin than Tabriz females. These results suggest the possibility of using selective crosses between wasp populations, in combination with artificial selection in the laboratory, to tune the thermal phenotype of parasitoids to specific regions prior to augmentative releases.

Drosophila suzukii (Spotted Wing Drosophila) has recently become a serious invasive pest of fruit crops in the USA, Canada, and Europe, leading to substantial economic losses. D. suzukii is a direct pest, ovipositing directly into ripe or ripening fruits; in contrast, other Drosophilids utilize decaying or blemished fruits and are nuisance pests at worst. Immature stages of D. suzukii are difficult to differentiate from other Drosophilids, posing problems for research and for meeting quarantine restrictions designed to prevent the spread of this pest in fruit exports. Here we used a combined phylogenetic and bioinformatic approach to discover genetic markers suitable for a species diagnostic protocol of this agricultural pest. We describe a molecular diagnostic for rapid identification of single D. suzukii larva using multiplex polymerase chain reaction. Our molecular diagnostic was validated using nine different species of Drosophila for specificity and 19 populations of D. suzukii from different geographical regions to ensure utility within species.

Cultivated orchids are the most abundantly attacked by polyphagous mealybugs. This study documented how different density of mealybug Pseudococcus longispinus (Targioni Tozzetti) infestation is associated with a response of antioxidative systems of Phalaenopsis × hybridum ‘Innocence’. The degree of cell damage, estimated by electrolyte leakage measurement and the level of thiobarbituric acid reactive substances (TBARS), the content of pigments as well as the activity of antioxidative enzymes and proline level, as measurements of stress and stress compensation in moth orchid were examined. The highest electrolyte leakage (EL) value among samples from colonized plants was found in the orchids from series III (50 individuals/plant), whereas the lowest in the plants from series II (20 individuals/plant). The TBARS content reached the highest level at the lowest number of feeding insects (series I). Peroxidase activity toward guaiacol was significantly increased in series I (5 individuals/plant). The highest catalase activity was recorded in plants colonized by the highest number of scale insects (series III). Whereas, the highest value of proline was in series II. The content of individual photosynthetic pigments (chlorophyll a, chlorophyll b and carotenoids) in plant tissues did not vary significantly between control and colonized orchids. The results have not confirmed hypothesis that the increasing number of mealybugs occurring on plant enhanced plant physiological response. The degree of longtailed mealybug infestation on plants was positively correlated only with electrolyte leakage and catalase activity in leaf tissues.