Since 1980, mosquito breeding habitats in the Upper Rhine Valley were routinely treated with Bacillus thuringiensis var. israelensis (Bti). Bti is considered to significantly reduce the number of mosquitoes, and – especially when used in higher dosages – to be toxic to other Nematocera species, e.g. Chironomidae, which could be food sources for aerial feeding predators. To investigate direct and indirect effects of routine Bti treatment on food sources for aerial feeding predators, the availability of flying insects in treated and untreated areas was compared. A car trap was used for insect collection, which allowed their exact spatiotemporal assignment. The statistical analysis revealed that insect taxa abundance was influenced differently by the factors season, temperature and time of day. Nematocera (Diptera) were the most frequently collected insects in all areas. Chironomidae were the predominant aquatic Nematocera. The comparison of treated and untreated sites did not show significant differences that would indicate any direct or indirect effect of routine Bti treatment on the availability of flying insects. Additional to food availability, food selection must be considered when investigating food resources for aerial feeding predators. In this study, food selection of Delichon urbicum (House Martin) as an example was investigated with the help of neck ring samples. The preferred prey of the investigated D. urbicum colony consisted of diurnal insects with terrestrial larvae (Aphidina, Brachycera, Coleoptera). Chironomidae were consumed, but not preferred.

Macdunnoughia crassisigna Warren (Lepidoptera: Noctuidae) is a highly destructive herbivore that poses a serious risk to cotton, maize, soybean, and cruciferous vegetables in East Asia. Examining the effects of various biotic and abiotic factors on the flight performance of M. crassisigna is crucial for a better understanding of its trans-regional migration. In this study, the flight activity of M. crassisignai moths of different ages, under different temperatures and relative humidity (RH) levels, was evaluated by tethering individuals to computerized flight mills for a 24-h trial period. The results showed that M. crassisignai had the capacity for sustained flight and the flight ability was strongest in 3-day-old individuals, and then their flight performance decreased significantly in older moths. For both sexes, temperature had a significant effect on their flight performance, and the flight activity was relatively higher at 24–28°C than other temperatures. There was a significant effect of RH on all flight parameters of the tested moths, and the flight activity was relatively higher at RH of 60–75% than other RH levels. For 3-day-old moths under the optimum conditions (24°C and 75% RH) throughout the 24 h scotophase, their mean flight distance reached 66 km, and the mean flight duration reached 13.5 h, suggesting M. crassisigna possess strong potential to undertake long-distance migration. These findings will be helpful for developing sound forecasting systems of this pest species.

Adulticides applied against mosquitoes can reduce vector populations during times of high arbovirus transmission. However, impacts of these insecticides on pollinators and other non-target organisms are of concern to mosquito control professionals, beekeepers and others. We evaluated mortality of Culex quinquefasciatus and Apis mellifera when caged insects were exposed to low and high label rates of four common adulticides (Aqua-Pursuit™ [permethrin], Duet® [prallethrin + sumithrin], Fyfanon® [malathion] and Scourge® [resmethrin]) at six distances up to 91.4 m from a truck-mounted ultra-low-volume sprayer. Honey bee mortality was both absolutely low (<10%) and low relative to mosquito mortality for most products, distances, and application rates. Exceptions were at the high rate of Fyfanon (honey bee mortality of 22–100% at distances ≤61 m) and the low rate of Scourge (mortality <10% for both insects). The greatest ratios of mosquito-to-honey bee mortality were found for the low rate of Fyfanon (30× greater) and the high rate of Duet (50× greater). Aqua-Pursuit and Fyfanon tended to increase mortality of both species at closer distances and at higher application rate; this was related to increased number and size of spray droplets. Wind speed and temperature had inconsistent effects on mortality of mosquitoes only. In this bioassay designed to have insects directly intercept insecticide droplets, mosquito adulticides applied at low rates and at >61 m had limited impacts on honey bee mortality while providing effective mosquito control.

Detrimental effects of ultraviolet (UV) light on living organisms are well understood, little is known about the effects of blue light irradiation. Although a recent study revealed that blue light caused more harmful effects on insects than UV light and blue light irradiation killed insect pests of various orders including Diptera, the effects of blue light on physiology of insects are still largely unknown. Here we studied the effects of blue light irradiation on cuticular melanin in larval and the immune response in adult stage of Bactrocera dorsalis. We also evaluated the effects of blue light exposure in larval stage on various age and mass at metamorphosis and the mediatory role of cuticular melanin in carryover effects of larval stressors across metamorphosis. We found that larvae exposed to blue light decreased melanin contents in their exoskeleton with smaller mass and delayed metamorphosis than insects reared without blue light exposure. Across metamorphosis, lower melanotic encapsulation response and higher susceptibility to Beauveria bassiana was detected in adults that had been exposed to blue light at their larval stage, thereby constituting the first evidence that blue light impaired adult immune function in B. dorsalis as a carryover effect of larval exposure.

Lymantia dispar (L.) (Lepidoptera: Lymantriidae), commonly known as the gypsy moth, is a serious forest pest, and beneficial insects are particularly important for reducing its population numbers. Ooencyrtus kuvanae (Howard) (Hymenoptera: Encyrtidae) is an arrhenotokous, solitary egg parasitoid of L. dispar. In this study, we evaluated a new substitute host, Philosamia ricini (Danovan) (Lepidoptera: Saturniidae) for O. kuvanae. We investigated some of the biological effects of O. kuvanae on P. ricini eggs. In this context, the importance of the age of the female parasitoid (1, 3 or 5 days old), host age (1–2 and 3–4 days old) and host number (40, 60 and 80 host eggs) were examined under laboratory conditions (25 ± 1 °C, 65 ± 5% relative humidity and a 16 : 8 h photoperiod [light : dark]). The highest rate of offspring production (89.90%) occurred with 40 (1–2-day-old) host eggs and 5-day-old females. The mean developmental period ranged from 16.5 ± 0.08 days to 18.7 ± 0.08 days. The mean lifespan of the parasitoid was 51.10 ± 1.1 (n = 60) days with bio-honey and 3.92 ± 0.14 (n = 60) days without food. The mean fecundity was 68.88 ± 3.22 offspring/female. Peak adult emergence occurred between 2 and 9 days. The mean oviposition and mean post-oviposition periods of the female parasitoid were 22.76 ± 1.37 days and 13.64 ± 1.40 days, respectively. O. kuvanae was reared for more than ten generations on the eggs of P. ricini. Based on our findings, P. ricini can be used to rear O. kuvanae for the biological control of L. dispar.

The Asian long-horned beetle (ALB), Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae: Lamiinae), is a wood-borer and polyphagous xylophage that is native to Asia. It infests and seriously harms healthy trees, and therefore is a cause for considerable environmental concern. The analysis of population genetic structure of ALB and sibling species Anoplophora nobilis (Ganglbauer) will not only help to clarify the relationship between environmental variables and mechanisms of speciation, but also will enhance our understanding of evolutionary processes. However, the known genetic markers, particularly microsatellites, are limited for this species. SSRLocator software was used to analyze the distribution and frequencies of genomic simple sequence repeat (SSR), to infer the basic characteristics of repeat motifs, and to design primers. We developed SSR loci of 2–6 repeated units, including 10,650 perfect SSRs, and found 140 types of repeat motifs. A total of 2621 SSR markers were discovered in ALB whole-genome shotgun sequences. 48 pairs of SSR primers were randomly chosen from 2621 SSR markers, and half of these 48 pairs were polymorphic containing 4 di-, 7 tri-, 2 tetra-, and 11-hexamer SSRs. Four populations test the effectiveness of the primers. These results suggest that our method for whole-genome SSR screening is feasible and efficient, and the SSR markers developed in this study are suitable for further population genetics studies of ALB. Moreover, they may also be useful for the development of SSRs for other Coleoptera.

The optimal use of available host by parasitoid insects should be favoured by natural selection. For solitary parasitoids, superparasitism (i.e. the egg-laying of several eggs/host) may represent a detrimental phenomenon both in a biological and an applied sense, but under certain circumstances it may be adaptive. Here, we studied the effects of increasing levels of superparasitism (LSPs, number of parasitoid larvae/host) on fitness-related parameters of the immature and adult stages of Diachasmimorpha longicaudata, a solitary endoparasitoid parasitizing Ceratitis capitata. We investigated the moment when supernumerary parasitoid larvae are eliminated and the effects produced by this process, together with its repercussion on female fecundity, parasitism rate, sex ratio, adult survival, flight ability and body size. Complete elimination of competitors occurred soon after larval hatching, before reaching the second larval stage. Elimination process took longer at higher LSPs, although a normal developmental (egg–adult) time was achieved. For LSPs 1, 2, 3 and 5 the effects on parasitoid emergence were mild, but LSP 10 led to the death of all developing parasitoids. Aside from this, to develop in superparasitized hosts did not significantly affect any of the evaluated parameters, and only a female-biased sex ratio was observed at higher LSPs. However, the effects of superparasitism on the adults may have a different outcome under more variable conditions in the field, once they are released for biological control purposes.

Whitefly, Bemisia tabaci, is one of the most important pests of tomato, Solanum lycopersicum L. The Mi-1 gene mediates tomato resistance to the Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) species of B. tabaci, three species of root-knot nematodes, Meloidogyne spp., and the potato aphid, Macrosiphum euphorbiae. Tomato seedlings bearing the Mi-1 gene are resistant to nematodes soon after germination but resistance to aphids is developmentally regulated; a reliable conclusion about Mi-1 resistance to B. tabaci was not available to date. In the present work, 3-, 5- and 8-week-old plants of the tomato cultivars Motelle and Moneymaker (bearing and lacking the Mi-1 gene, respectively) were simultaneously tested under free-choice (antixenosis) and no-choice (antibiosis) conditions, to assess the real influence of plant age on the Mi-1-mediated resistance to the MED species of B. tabaci. Subsequently, plants of the same age but with different level of development were compared to check whether the plant size can also affect this tomato resistance. Obtained results demonstrated that Mi-1-mediated resistance to B. tabaci is developmentally regulated, as variations in the age of bearing-Mi-1 plants affects most infestation parameters tested. Differences between cultivars with and without the Mi-1 gene were significant for 8- but not for 3-week-old plants. For 5-week-old plants, differences between cultivars were less pronounced than in older plants, expressing an intermediate level of resistance in Motelle. Plant size also influenced whitefly infestation and reproductive activity on the resistant cultivar. However, plant age has more impact than plant size on the Mi-1-mediated resistance of tomato to B. tabaci.

Helicoverpa armigera is a significant agricultural pest and particularly notorious for its resistance to many types of common insecticides. RNA interference (RNAi) is a mechanism of post-transcriptional gene silencing and trigged by double-strand RNA (dsRNA), has become a widely used reverse genetics and potent tool for insect pest control. In this study, the effect of ingestion and injection delivery methods of dsRNA related two important enzyme genes, α-amylase (HaAMY48, Ha-AMY49) and juvenile hormone esterase (Ha-JHE), were examined on growth and development of H. armigera. After 24, 48, 72 and 96 h of feeding bioassay, significant down regulation was observed about; 56, 68, 78, 80.75% for HaAMY48, 60, 70, 86.5 and 96.75%, for Ha-AMY49 and 14, 27.5, 23 and 31.7% for Ha-JHE, respectively. The results for injection assay was 61.5, 71.5, 74 and 95.8% for Ha-AMY48; 70, 88, 91.5 and 97.7% for Ha-AMY49 and 22, 61, 75 and 74% for Ha-JHE after 24, 48 and 72 h of last injecting, respectively. Larvae that treated with dsRNA, fed or injected, lost more than half of their weight. 50% mortality in treated larvae was observed in the case injection bioassay with dsHa-JHE and 59% of larvae that fed of dsRNA-treated cubes survived. DsHa-AMY48 and 49 have significant mortality, but mixing of them is more effective in both bioassays. Injection bioassay has a potent inhibitory effect on α-amylase-specific activity about more than 87% in treated larvae with mix of dsHa-AMY48 and 49. Adult malformation percent was evaluated for feeding (28, 35.5 and 43% for Ha-AMY48, 49 and Ha-JHE, respectively) and injection bioassay (23, 42 and 29% for Ha-AMY48, 49 and Ha-JHE, respectively). All these finding suggest that Ha-AMY48, Ha-AMY49 and Ha-JHE can be new candidates to scheming effective dsRNAs pesticide for H. armigera control.

Egg parasitoid rearing on factitious hosts is an important step to reduce costs and increase availability of the biocontrol agent but it requires quality control to achieve success in field conditions. To this end, this study evaluated the quality of Telenomus remus (Hymenoptera: Platygastridae) reared on Corcyra cephalonica (Lepidoptera: Pyralidae) for until 45 generations. In the first bioassay, we evaluated the body size of the laboratory-produced parasitoids. In the second bioassay, flight activity was examined, measuring the percentage of ‘flyers’, ‘walkers’ and ‘deformed’ parasitoids. The third bioassay assessed parasitism on Spodoptera frugiperda (Lepidoptera: Noctuidae) eggs. Our data indicate that the laboratory-reared parasitoid neither lost its ability to fly nor to parasitize S. frugiperda eggs. In conclusion, quality did not decrease significantly during 45 generations, and therefore rearing of T. remus on C. cephalonica as factitious host promises to be successful.

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.

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.

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.

Treating plants with synthetic jasmonic acid (JA) induces a defensive response similar to herbivore attack, and is a potential strategy for integrated pest management. Despite the importance of sugarcane, its JA-induced defences have not yet been studied. We investigated the effects of JA treatment on the direct and indirect resistance of sugarcane to the key-pest and specialist herbivore Diatraea saccharalis and the generalist Spodoptera frugiperda. Indirect defences were examined by testing the attraction of Cotesia flavipes, a sugarcane-borer parasitoid, to JA-induced volatile. The results showed that JA-treated sugarcane did not affect the weight gain of the two larvae. However, in dual-choice assays, both species preferred to feed on mock rather than JA-treated plants. Leaf colorimetric analyses showed that visual cues are unlikely to be involved in larval preference, whereas results from olfactometric assays revealed that D. saccharalis preferred JA-induced over mock plant volatiles. After 48 h of treatment, JA-treated plants emitted a volatile blend attractive to C. flavipes, comprised mainly of sesquiterpenes. However, the parasitoid did not discriminate JA-treated from host-damaged plant volatiles. When the wasps were given a choice between JA-treated and JA-treated + host-damaged plants, they preferred the latter, which emitted a more complex blend, suggesting that JA treatment likely does not hamper host-finding. We concluded that JA induces the emission of volatiles that are attractive to the sugarcane borer parasitoid, as well as an antixenosis type of resistance in sugarcane against the two pests, although neither volatiles nor visual cues alone are involved in the underlying mechanism.

The E2F transcription factor family is distributed widely in eukaryotes and has been well studied among mammals. In the present study, the E2F transcription factor 4 (E2F4) gene was isolated from fat bodies of Antheraea pernyi and sequenced. E2F4 comprised a 795 bp open reading frame encoding a deduced amino acid sequence of 264 amino acid residues. The recombinant protein was expressed in Escherichia coli (Transetta DE3), and anti-E2F4 antibodies were prepared. The deduced amino acid sequence displayed significant homology to an E2F4-like protein from Bombyx mori L. Quantitative real-time polymerase chain reaction analysis revealed that E2F4 expression was highest in the integument, followed by the fat body, silk glands, and haemocytes. The expression of E2F4 was upregulated in larvae challenged by bacterial (Escherichia coli, Micrococcus luteus), viral (nuclear polyhedrosis virus), and fungal (Beauveria bassiana) pathogens. These observations indicated that E2F4 is an inducible protein in the immune response of A. pernyi and probably in other insects.