Chemical insecticides have been widely used to control insect pests, leading to the selection of resistant populations. To date, several single nucleotide polymorphisms (SNPs) have already been associated with insecticide resistance, causing reduced sensitivity to many classes of products. Monitoring and detection of target-site resistance is currently one of the most important factors for insect pest management strategies. Several methods are available for this purpose: automated and high-throughput techniques (i.e. TaqMan or pyrosequencing) are very costly; cheaper alternatives (i.e. RFLP or PASA–PCRs) are time-consuming and limited by the necessity of a final visualization step. This work presents a new approach (QSGG, Qualitative Sybr Green Genotyping) which combines the specificity of PASA–PCR with the rapidity of real-time PCR analysis. The specific real-time detection of Cq values of wild-type or mutant alleles (amplified used allele-specific primers) allows the calculation of ΔCqW–M values and the consequent identification of the genotypes of unknown samples, on the basis of ranges previously defined with reference clones. The methodology is applied here to characterize mutations described in Myzus persicae and Musca domestica and we demonstrate it represents a valid, rapid and cost-effective technique that can be adopted for monitoring target-site resistance in field populations of these and other insect species.

In tritrophic ‘plants–herbivores–natural enemies’ systems, there are relatively few reports concerning the role(s) of kairomones in pupal parasitism. Chouioia cunea Yang (Hymenoptera: Eulophidae), an endoparasitic chalcid wasp, parasitizes pupae of the fall webworm (Hyphantria cunea Drury). The role of host-related kairomones was investigated using electroantennogram (EAG) and behavioral techniques. Chemicals from some host stages (pupae) and host by-products (frass), induced arrestment behavior of female parasitoids, while chemicals from prepupae, were inactive. Gas chromatography–mass spectrometry analysis of volatiles collected from pupae, frass and prepupae using solid-phase microextration revealed seven compounds with carbon chain lengths ranging from C4 to C20. All of the chemicals elicited significant EAG responses in C. cunea. Y-tube olfactometer bioassays demonstrated a significant positive response of mated female C. cunea to 1-dodecene. These data provide a better understanding of the host location mechanisms of pupal parasitoid.

In urban landscapes, gardens provide refuges for bee diversity, but conservation potential may depend on local and landscape features. Foraging and population persistence of bee species, as well as overall pollinator community structure, may be supported by the abundance, richness, and spatial distribution of floral resources. Floral resources strongly differ in urban gardens. Using hand netting and pan traps to survey bees, we examined whether abundance, richness, and spatial distribution of floral resources, as well as ground cover and garden landscape surroundings influence bee abundance, species richness, and diversity on the central coast of California. Differences in floral abundance and spatial distribution, as well as urban cover in the landscape, predicted different bee community variables. Abundance of all bees and of honeybees (Apis mellifera) was lower in sites with more urban land cover surrounding the gardens. Honeybee abundance was higher in sites with patchy floral resources, whereas bee species richness and bee diversity was higher in sites with more clustered floral resources. Surprisingly, bee species richness and bee diversity was lower in sites with very high floral abundance, possibly due to interactions with honeybees. Other studies have documented the importance of floral abundance and landscape surroundings for bees in urban gardens, but this study is the first to document that the spatial arrangement of flowers strongly predicts bee abundance and richness. Based on these findings, it is likely that garden managers may promote bee conservation by managing for floral connectivity and abundance within these ubiquitous urban habitats.

The endoparasitoid Microplitis prodeniae Rao and Chandry is an important potential augmentative biological control agent for lepidopteran pests of vegetables and tobacco. However, cold storage of pupae is required to ensure that sufficient parasitoids are available when they are needed in the field. In this study, pupae were maintained at 0, 4 or 10°C for 5–50 days after which the adults were evaluated for emergence, pre-emergence period, sex ratio, female longevity, oviposition period, and fecundity. Cold storage did not affect the pre-emergence period or proportion of females; however, there was a significant reduction in emergence, female longevity, oviposition period, and fecundity with increased exposure to cold. The pre-emergence period was approximately 5 days, and approximately 50% of the emergent parasitoids were females. A cold storage regime of 10 days at 10°C had no effect on the parasitoids and adult emergence was greater than 50% even after 20 days at 10°C. There was no carryover of the cold treatment from parental to F1 and F2 generations. Thus, M. prodeniae can be stockpiled for field release by exposing the pupae to a cold regime and subsequently holding them for adult emergence at 28°C.

Cerambyx cerdo (Cc) is a protected saproxylic beetle in Europe, although it is increasingly reported as an oak ‘pest’. Cc ecological features are relatively well known, but, its reproductive biology is still poorly understood. Hence, we investigated the reproductive traits of Cc under laboratory conditions. In females, body length was 44.1 ± 0.9 mm, 28–53 (mean ± SE, range); fecundity 143 ± 11 eggs, 33–347; fertility 78 ± 1%, 65–93; oviposition period 44 ± 3 days, 13–128 and longevity 59 ± 5 days, 16–157. Fecundity was positively correlated with female size, longevity and oviposition period. Daily fecundity was 3.5 ± 0.2 eggs/day, 0.9–6.5 showing a fluctuating synovigenic pattern with a slight decreasing trend over time. Egg length was 3.74 ± 0.01 mm, 2.3–6.0 and egg volume 5.45 ± 0.04 mm3, 2.4–9.6. Egg size was correlated with female size, but, the relative size of eggs was larger in smaller females. Incubation time was 13.5 ± 0.1 days, 7–28. Hatching was superior in larger eggs and neonate size was positively correlated to egg volume. Females were polyandrous (up to 19 matings), but, multiple mating did not enhance fecundity or fertility. In males, body length was 41.8 ± 0.8 mm, 29–53 and longevity 49 ± 3 days, 9–124. Male longevity was unrelated to body size. Males were polygynous (up to 16 matings) and mating number did not affect male longevity. Overall, females were larger and lived longer than males. Cc reproductive traits are compared with those other Cerambycidae, especially with the congeneric pest Cerambyx welensii. Our data may be valuable to improve the protection/management measures of Cc in dehesa woodlands and other oak forests.

The oriental army worm Mythimna separata (Lepidoptera: Noctuidae) is a migratory pest in Eastern Asia and China. Seasonal high temperatures in Southern China and low temperatures in Northern China are pressures favouring the annual migration of this species, while cold tolerance determines the northern limit of its overwintering range. A number of physiological stress responses occur in insects as a result of variations in temperature. One reaction to thermal stress is the generation of reactive oxygen species (ROS), which can be harmful by causing oxidative damage. The time-related effects (durations of 1, 4 and 7 h) of thermal stress treatments of M. separata at comparatively low (5, 10, 15 and 20°C) and high (30, 35, 40 and 45°C) temperatures on the activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), peroxidase (POX) and glutathione S-transferases (GSTs), and total antioxidant capacity (T-AOC) were determined. Thermal stress resulted in significant elevation of the activities of SOD, CAT and GSTs, indicating that these enzymes contribute to defence mechanisms counteracting oxidative damage caused by an increase in ROS. However, at high-temperatures, POX and T-AOC were also found to contribute to scavenging ROS. Our results also indicate that extreme temperatures lead to elevated ROS production in M. separata. The present study confirms that thermal stress can be responsible for oxidative damage. To overcome such stress, antioxidant enzymes play key roles in diminishing oxidative damage in M. separata.

In this study, we investigated thermotolerance, several physiological responses and damage to reproductive cells in chlorpyrifos-resistant (Rc) and -susceptible (Sm) strains of the diamondback moth, Plutella xylostella subjected to heat stress. The chlorpyrifos resistance of these strains was mediated by a modified acetylcholinesterase encoded by an allele, ace1R, of the ace1 gene. Adults of the Rc strain were less heat resistant than those of the Sm strain; they also had lower levels of enzymatic activity against oxidative damage, higher reactive oxygen species contents, weaker upregulation of two heat shock protein (hsp) genes (hsp69s and hsp20), and stronger upregulation of two apoptotic genes (caspase-7 and -9). The damage to sperm and ovary cells was greater in Rc adults than in Sm adults and was temperature sensitive. The lower fitness of the resistant strain, compared with the susceptible strain, is probably due to higher levels of oxidative stress and apoptosis, which also have deleterious effects on several life history traits. The greater injury observed in conditions of heat stress may be due to both the stronger upregulation of caspase genes and weaker upregulation of hsp genes in resistant than in susceptible individuals.

The European winter moth, Operophtera brumata, is a non-native pest in the Northeastern USA causing defoliation of forest trees and crops such as apples and blueberries. This species is known to hybridize with O. bruceata, the Bruce spanworm, a native species across North America, although it is not known if there are hybrid generations beyond F1. To study winter moth population genetics and hybridization with Bruce spanworm, we developed two sets of genetic markers, single nucleotide polymorphisms (SNPs) and microsatellites, using genomic approaches. Both types of markers were validated using samples from the two species and their hybrids. We identified 1216 SNPs and 24 variable microsatellite loci. From them we developed a subset of 95 species-diagnostic SNPs and ten microsatellite loci that could be used for hybrid identification. We further validated the ten microsatellite loci by screening field collected samples of both species and putative hybrids. In addition to confirming the presence of F1 hybrids reported in previous studies, we found evidence for multi-generation asymmetric hybridization, as suggested by the occurrence of hybrid backcrosses with the winter month, but not with the Bruce spanworm. Laboratory crosses between winter moth females and Bruce spanworm males resulted in a higher proportion of viable eggs than the reciprocal cross, supporting this pattern. We discuss the possible roles of population demographics, sex chromosome genetic incompatibility, and bacterial symbionts as causes of this asymmetrical hybridization and the utility of the developed markers for future studies.

The importance of leaf surface wax compounds from the rice-field weed Ludwigia octovalvis (Jacq.) Raven (Onagraceae) was determined in the flea beetle Altica cyanea (Weber) (Coleoptera: Chrysomelidae). Extraction, thin layer chromatography and GC-MS and GC-FID analyses of surface waxes of young, mature and senescent leaves revealed 20, 19 and 19 n-alkanes between n-C15 and n-C35, respectively; whereas 14, 14 and 12 free fatty acids between C12:0 and C22:0 fatty acids were identified in young, mature and senescent leaves, respectively. Tricosane was predominant n-alkane in young and mature leaves, whilst eicosane predominated in senescent leaves. Heneicosanoic acid, palmitic acid and docosanoic acid were the most abundant free fatty acids in young, mature and senescent leaves, respectively. A. cyanea females showed attraction to 0.25 mature leaf equivalent surface waxes compared with young or senescent leaves in a short glass Y-tube olfactometer bioassay. The insects were attracted to a synthetic blend of 0.90, 1.86, 1.83, 1.95, 0.50 and 0.18 µg ml−1 petroleum ether of hexadecane, octadecane, eicosane, tricosane, palmitic acid and alpha-linolenic acid, respectively, comparable with the proportions as present in 0.25 mature leaf equivalent surface waxes. A. cyanea also laid eggs on a filter paper moistened with 0.25 mature leaf equivalent surface waxes or a synthetic blend of 0.90, 1.86, 1.83, 1.95, 0.50 and 0.18 µg ml−1 petroleum ether of hexadecane, octadecane, eicosane, tricosane, palmitic acid and alpha-linolenic acid, respectively. This finding could provide a basis for monitoring of the potential biocontrol agent in the field.

Changes in environmental conditions influence the performance of organisms in every aspect of their life. Being capable of accurately sensing these changes allow organisms to better adapt. The detection of environmental conditions involves different sensory modalities. There are many studies on the morphology of different sensory structures but not so many studies showing their function. Here we studied the morphology of different sensory structures in the larva of a dipteran parasitoid. We occluded the putative sensory structures coupling the morphology with their function. First, we could develop a non-invasive method in which we occluded the putative sensorial structures annulling their function temporarily. Regarding their functionality, we found that larvae of Mallophora ruficauda require simultaneously of the sensilla found both in the antennae and those of the maxillary palps in order to orient to its host. When either both antennae or both maxillary palps were occluded, no orientation to the host was observed. We also found that these structures are not involved in the acceptance of the host because high and similar proportion of parasitized hosts was found in host acceptance experiments. We propose that other sensilla could be involved in host acceptance and discuss how the different sensilla in the antennae and maxillary palps complement each other to provide larvae with the information for locating its host.

In laboratories, the parasitism rate of Ostrinia furnacalis (Güenée) (Lepidoptera: Crambidae) eggs by Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae) is low; however, efforts to control O. furnacalis with T. dendrolimi in the field have been successful. In this study, the effects of the number of attacks by T. dendrolimi against O. furnacalis eggs and diet of O. furnacalis larva on wasp development were investigated. The results indicated that more attacks increased significantly not only the successful parasitism rate of O. furnacalis eggs by T. dendrolimi, but also the percentage of host eggs that failed to develop into either O. furnacalis larvae or T. dendrolimi. Both the size and female proportion of T. dendrolimi offspring decreased as the number of attacks increased. The number of T. dendrolimi eggs laid in per host egg increased significantly as the ratio of wasps to host eggs increased from 1:5 to 3:5. Host diet also significantly affected the developmental time of immaturity and the emergence rate of adults of T. dendrolimi. These results illustrate how inundative releases of T. dendrolimi can successfully control O. furnacalis despite the fact that pest parasitism by the subsequent wasp generation decreases sharply in the field. The suitability of O. furnacalis eggs to T. dendrolimi and the superparasitism effects on offspring of T. dendrolimi are discussed.

Traps designed to capture insects during normal movement/dispersal, or via attraction to non-specific (plant) volatile lures, yield by-catch that carries valuable information about patterns of community diversity and composition. In order to identify potential native/introduced pests and detect predictors of colonization of non-native pines, we examined beetle assemblages captured in intercept panel traps baited with kairomone lures used during a national monitoring of the woodwasp, Sirex noctilio, in Southern Africa. We identified 50 families and 436 morphospecies of beetles from nine sites sampled in both 2008 and 2009 and six areas in 2007 (trap catch pooled by region) across a latitudinal and elevational gradient. The most diverse groups were mainly those strongly associated with trees, known to include damaging pests. While native species dominated the samples in terms of richness, the dominant species was the introduced bark beetle Orthotomicus erosus (Curculionidae: Scolytinae) (22 ± 34 individuals/site). Four Scolytinae species without previous records in South Africa, namely Coccotrypes niger, Hypocryphalus robustus (formerly Hypocryphalus mangiferae), Hypothenemus birmanus and Xyleborus perforans, were captured in low abundances. Communities showed temporal stability within sites and strong biogeographic patterns across the landscape. The strongest single predictors of community composition were potential evaporation, latitude and maximum relative humidity, while the strongest multifactor model contained elevation, potential evaporation and maximum relative humidity. Temperature, land use variables and distance to natural areas did not significantly correlate with community composition. Non-phytophagous beetles were also captured and were highly diverse (32 families) perhaps representing important beneficial insects.

The occurrence of pesticidal pollution in the environment and the resistance in the mosquito species makes an urge for the safer and an effective pesticide. Permethrin, a poorly water-soluble pyrethroid pesticide, was formulated into a hydrodispersible nanopowder through rapid solvent evaporation of pesticide-loaded oil in water microemulsion. Stability studies confirmed that the nanopermethrin dispersion was stable in paddy field water for 5 days with the mean particle sizes of 175.3 ± 0.75 nm and zeta potential of −30.6 ± 0.62 mV. The instability rate of the nanopermethrin particles was greater in alkaline (pH 10) medium when compared with the neutral (pH 7) and acidic (pH 4) dispersion medium. The colloidal dispersion at 45°C was found to be less stable compared with the dispersions at 25 and 5°C. The 12- and 24-h lethal indices (LC50) for nanopermethrin were found to be 0.057 and 0.014 mg l−1, respectively. These results were corroborative with the severity of damages observed in the mosquito larvae manifested in epithelial cells and the evacuation of the midgut contents. Further, the results were substantiated by the decrease in cellular biomolecules and biomarker enzyme activity in nanopermethrin treated larvae when compared to bulk and control treatment.

Two bioassays were conducted in parallel to assess the effects of cattle treated with either 1% ivermectin (IVM) or 3.15% IVM (dosed at 0.2 and 0.63 mg kg−1, respectively) on reproduction and survival of Onthophagus landolti Harold. Adult beetles were exposed 10 days to faeces of treated cattle starting at: one day before treatment (controls), 3, 6, 14, 28 and 35 days post-treatment. Adult survival of O. landolti was not affected by either of the two treatments. Faecal residues of 1% IVM almost completely suppressed fecundity of beetles at 3, 6 and 14 days post-treatment (dPT), and reduced fecundity of O. landolti at 28 dPT ( 38.3%), relative to controls. Meanwhile, IVM residues after treatment with 3.15% IVM almost completely suppressed fecundity of beetles at 3, 6, 14 and 28 dPT, and reduced fecundity of O. landolti at 35 dPT (80.9%), relative to controls. Larval survival was significantly reduced only at 3 dPT with 1% IVM. Meanwhile, treatment with 3.15% IVM significantly reduced larval survival at 6, 14 and 28 dPT. Larval mortality was recorded only in L-I and L-II instars. Moreover, in both bioassays, most of the L-I and L-II specimens that survived showed signs of toxicity. In conclusion, residual IVM in cattle faeces after treatment with injectable IVM has a detrimental effect on the fecundity of adult O. landolti up to 4 weeks post-treatment and on the subsequent larval survival.

The objective of this study was to evaluate the enzymatic activity of homogenates of insects fed on grain of cowpea, Vigna unguiculata (L.), cultivars grown with different nitrogen sources. For the experiment we used aliquots of the homogenate of 100 unsexed adult insects, emerged from 10 g of grain obtained from four cowpea cultivars: ‘BRS Acauã’, ‘BRS Carijó’, ‘BRS Pujante’, and ‘BRS Tapaihum’ grown under different regimes of nitrogen sources: mineral fertilizer, inoculation with strains of diazotrophs (BR 3267, BR 3262, BR 3299; INPA 03–11B, 03–84 UFLA, as well as the control (with soil nitrogen). The parameters evaluated were enzymatic activities of insect protease, amylase and lipase and the starch content of the grains. There were differences in the enzymatic activity of amylase, lipase and protease of insect homogenate according to the food source. A lower activity of the enzyme amylase from C. maculatus homogenate was observed when insects were fed grain of the cultivar BRS Carijó. A lower activity of lipase enzyme from C. maculatus homogenate was observed when the insects fed on grain from the interaction of the cultivar Tapaihum inoculated with BR 3262 diazotrophs. The lowest proteolytic activity was observed in homogenate of insects fed on interaction of ‘BRS Carijó’ inoculated with BR 3262 diazotrophs. Starch content correlated positively with the amylase activity of C. maculatus homogenate. The cultivar BRS Carijó had a different behavior from the other cultivars, according to the cluster analysis.

Oedaleus asiaticus Bey. Bienko is a significant grasshopper pest species occurring in north Asian grasslands. Outbreaks often result in significant loss in grasses and economic losses. Interestingly, we found this grasshopper was mainly restricted to Stipa-dominated grassland. We suspected this may be related to the dominant grasses species, Stipa krylovii Roshev, and hypothesized that S. krylovii contributes to optimal growth performance and population distribution of O. asiaticus. A 4 year investigation showed that O. asiaticus density was positively correlated to the above-ground biomass of S. krylovii and O. asiaticus growth performance variables (survival rate, size, growth rate) were significantly higher in Stipa-dominated grassland. A feeding trial also showed that O. asiaticus had a higher growth performance when feeding exclusively on S. krylovii. In addition, the choice, consumption and the efficiency of conversion of ingested food (ECI) by O. asiaticus was highest for S. krylovii compared with other plant species found in the Asian grasslands. These ecological and biological traits revealed why O. asiaticus is strongly associated with Stipa-dominated grasslands. We concluded that the existence of S. krylovii benefited the growth performance and explained the distribution of O. asiaticus. These results are useful for improved pest management strategies and developing guidelines for the monitoring of grasshopper population dynamics against the background of vegetation succession and changing plant communities in response to activities such as grazing, fire and climate change.

Although some novel antimicrobial peptides (AMP) have been successfully isolated from Bactrocera dorsalis Hendel, the mechanisms underlying the induction of these peptides are still elusive. The homolog of NF-κB transcription factor Relish, designated as BdRelish, was cloned from B. dorsalis. The full length cDNA of BdRelish is 3954 bp with an open reading frame that encodes 1013 amino acids. Similar to Drosophila Relish and the mammalian p100, it is a compound protein containing a conserved Rel homology domain, an IPT (Ig-like, plexins, transcription factors) domain and an IκB-like domain (four ankyrin repeats), the nuclear localization signal RKRRR is also detected at the residues 449–453, suggesting that it has homology to Relish and it is a member of the Rel family of transcription activator proteins. Reverse transcription quantitative polymerase chain reaction analysis reveals that BdRelish mRNAs are detected in different quantities from various tissues and the highest transcription level of BdRelish is determined in fat body. The injection challenge of Escherichia coli and Staphylococcus aureas significantly upregulated the expression of BdRelish. The injection of BdRelish dsRNA markedly reduced the expression of BdRelish and decreased the transcription magnitude of antimicrobial peptides. Individuals injected BdRelish dsRNA died at a significantly faster rate compared with the control groups. Therefore, BdRelish is vital for the transcription of AMPs to attack the invading bacteria.

The aim of this study was to evaluate the effects of silicon application and administration of the phytohormone gibberellic acid on resistance of the corn plants to the fall armyworm (FAW), Spodoptera frugiperda, and their vegetative characteristics. We evaluated larval and pupal duration, survival and biomass, and adult longevity, malformation and fecundity of S. frugiperda after feeding on plant matter treated with silicon and/or gibberellic acid. The feeding preference of FAW first-instar larvae, the total leaf area consumed by the insects, and the vegetative parameters of corn plants were also evaluated. No significant differences were observed in the measured parameters of larval and pupal stages of S. frugiperda in response to silicon or gibberellic acid. In adult stage insects, the number of eggs per female was significantly reduced in insects derived from larvae fed plants treated with silicon or gibberellic acid. In a non-preference test, 48 h after release, caterpillars preferred control untreated plants and consumed less matter from plants that had received hormonal treatment (gibberellic acid). Gibberellic acid also altered the vegetative characteristics of plants, by increasing their height, shoot fresh and dry mass, and silicon content. We conclude that gibberellic acid can alter the vegetative characteristics and silicon uptake of corn plants, leading to a reduction in their consumption by S. frugiperda larvae and a decrease in female insect oviposition.

Cosmopolitan pests such as Brevicoryne brassicae, Lipaphis pseudobrassicae, and Myzus persicae (Aphididae) cause significant damage to Brassicaceae crops. Assessment of the important biotic and abiotic factors that regulate these pests is an essential step in the development of effective Integrated Pest Management programs for these aphids. This study evaluated the influence of leaf position, precipitation, temperature, and parasitism on populations of L. pseudobrassicae, M. persicae, and B. brassicae in collard greens fields in the Triângulo Mineiro region (Minas Gerais state), Brazil. Similar numbers of B. brassicae were found on all parts of the collard green plants, whereas M. persicae and L. pseudobrassicae were found in greatest numbers on the middle and lower parts of the plant. While temperature and precipitation were positively related to aphid population size, their effects were not accumulative, as indicated by a negative interaction term. Although Diaeretiella rapae was the main parasitoid of these aphids, hyperparasitism was dominant; the main hyperparasitoid species recovered from plant samples was Alloxysta fuscicornis. Parasitoids seem to have similar distributions on plants as their hosts. These results may help predict aphid outbreaks and gives clues for specific intra-plant locations when searching for and monitoring aphid populations.

Heteroplasmy is the existence of multiple mitochondrial DNA haplotypes within the cell. Although the number of reports of heteroplasmy is increasing for arthropods, the occurrence, number of variants, and origins are not well studied. In this research, the occurrence of heteroplasmy was investigated in Thrips tabaci, a putative species complex whose lineages can be distinguished by their mitochondrial DNA haplotypes. The results from this study showed that heteroplasmy was due to the occurrence of mitochondrial cytochrome oxydase I (mtCOI) haplotypes from two different T. tabaci lineages. An assay using flow cytometry and quantitative real-time PCR was then used to quantify the per cell copy number of the two mtCOI haplotypes present in individuals exhibiting heteroplasmy from nine geographically distant populations in India. All of the T. tabaci individuals in this study were found to exhibit heteroplasmy, and in every individual the per cell copy number of mtCOI from lineage 3 comprised 75–98% of the haplotypes detected and was variable among individuals tested. There was no evidence to suggest that the presense of lineage-specific haplotypes was due to nuclear introgression; however, further studies are needed to investigate nuclear introgression and paternal leakage during rare interbreeding between individuals from lineages 2 and 3.