Diaphorina is a species-rich genus, native to the tropics and subtropics of the Old World, particularly of more arid regions. One of the species, Diaphorina citri, is the economically most important pest of citrus. Diaphorina species are morphologically similar which makes their identification difficult. In this study, the accuracy of DNA barcoding, using mitochondrial cytochrome c oxidase subunit 1 (COI), geometric morphometrics of the forewing and digital image processing methods were tested for identification of the three Diaphorina species: D. chobauti, D. citri and D. zygophylli. Moreover, the published COI sequences of D. citri, D. communis and D. lycii obtained from Genbank were used for cluster analyses. DNA barcodes for D. chobauti and D. zygophylli are deposited in Genbank for the first time. The results of the molecular and geometric morphometric analyses are congruent and place D. chobauti as the sister taxon of the other Diaphorina species. The geometric morphometric analysis shows that in D. zygophylli the fore margin is slightly curved proximally and sharply bent distally, while in D. chobauti and D. citri it is straight proximally and weakly bent distally. The results of digital image processing show that the distribution of the dark pattern differs consistently in the three studied species.

Plants not only respond to herbivorous damage but adjust their defense system after egg deposition by pest insects. Thereby, parasitoids use oviposition-induced plant volatiles to locate their hosts. We investigated the olfactory behavioral responses of Trichogramma pretiosum Riley, 1879 (Hymenoptera: Trichogrammatidae) to volatile blends emitted by maize (Zea mays L.) with singular and stacked events after oviposition by Spodoptera frugiperda Smith, 1797 (Hymenoptera: Trichogrammatidae) moths. Additionally, we examined possible variations in gene expression and on oviposition-induced volatiles. We used a Y-tube olfactometer to test for the wasp responses to volatiles released by maize plants oviposited by S. frugiperda and not-oviposited plants. Using the real-time PCR technique (qRT-PCR), we analyzed the expression of lipoxygenase and three terpene synthases genes, which are enzymes involved in the synthesis of volatile compounds that attract parasitoids of S. frugiperda. Olfactometer tests showed that T. pretiosum is strongly attracted by volatiles from transgenic maize emitted by S. frugiperda oviposition (VTPRO 3, more than 75% individuals were attracted). The relative expression of genes TPS10, LOX e STC was higher in transgenic hybrids than in the conventional (isogenic line) hybrids. The GC-MS analysis revealed that some volatile compounds are released exclusively by transgenic maize. This study provides evidence that transgenic hybrids enhanced chemical cues under oviposition-induction and helped to increase T. pretiosum efficiency in S. frugiperda control. This finding shows that among the evaluated hybrids, genetically modified hybrids can improve the biological control programs, since they potentialize the egg parasitoid foraging, integrating pest management.

Primary pests such as Rhyzoperta dominica may increase the contents of dockage, dust, and frass in grain mass. Although it has been suggested that frass can affect the population growth of stored product pests and ecological interactions among primary and secondary pests in stored grain, this has not been validated experimentally. Therefore, this work experimentally tested the hypothesis that R. dominica wheat frass may support population increases in secondary pests such as Tribolium confusum, T. castaneum, and Oryzaephilus surinamensis for the first time. The effect of frass on secondary pest performance was compared with the effects of various physical qualities of wheat grain (i.e., intact grain kernels, grain fragments, flour, grain + frass) and an artificially enriched control diet (milled wheat kernels, oat flakes, and yeast). The results showed that the clean intact grain kernels did not support the population growth of any tested species, and the nutrient-rich control diet provided the best support. Frass was a significantly better food medium for O. surinamensis and T. castaneum than flour or cracked grain, while T. confusum performed equally well on flour and frass. Our results showed that in terms of food quality and suitability for the tested species, frass occupied an intermediate position between the optimized breeding diet and simple uniform cereal diets such as cracked grain or flour. The results suggest that (i) the wheat frass of primary pest R. dominica is a riskier food source for the development of the tested secondary pests than intact or cracked wheat grain or flour; (ii) frass has the potential to positively influence interspecific interactions between R. dominica and the tested secondary pests; and (iii) wheat grain should be cleaned if increases in R. dominica populations and/or accumulated frass are detected.

The introduction of alien species is one of the main problems in conservation. Many successful invaders cause severe economic and ecological damage. Such is the case of Leptoglossus occidentalis, a phytophagous true bug native to North America, which has become a pest in Europe, Asia, Africa and South America. Within the genus, another species whose distributional range is expanding toward the east of North America is Leptoglossus clypealis. As climate determines the successful establishment of insects, the identification of climatically suitable areas for invasive species based on ecological niche models (ENMs) offers an excellent opportunity for preventing invasions. In this study, ENMs were built for both species and their native climatic niches were compared. Their niche breath was also measured. The climatic niches of both species are identical and the niche breadth of L. clypealis is broader than that of L. occidentalis. In view of the great ecological resemblance between these two species, we believe that L. clypealis could became a major pest thus it should be carefully monitored. The results of the present worldwide ENMs showed numerous regions with suitable conditions for the establishment of both species. The future ENMs exhibited a retraction in the suitable areas in North America, Europe and Asia.

Insecticide application and augmentative parasitoid releases are often considered incompatible. However, pesticide applications and parasitoid releases can be integrated into a pest management scheme if there is careful time scheduling of these interventions. In this study, we assessed the influence of commonly used insecticides (chlorpyrifos-methyl, deltamethrin, pyriproxyfen, thiamethoxam) in olive agroecosystems to two currently present Trichogramma parasitoids in the Mediterranean basin. Exposure to insecticides in relation to parasitoid's development was also tested. Both, insecticide type and application time influenced parasitism and the emergence rates of the two parasitoid species. Chlorpyrifos-methyl had the strongest impact on parasitoids resulting in low numbers of emerged adults followed by deltamethrin. The two parasitoids also exhibited different levels of susceptibility to the insecticides used. Potential integration of insecticides to integrated pest management using Trichogramma parasitoids is discussed.

Western flower thrips (WFT), Frankliniella occidentalis, is one of the most destructive pests of vegetables, fruits and ornamental crops worldwide, causing extensive damage by direct feeding of the crop and transmitting economically important viruses. Despite the successes of biocontrol agents to control WFT, more efficient and cost-effective ways must be found to encourage grower adoption of integrated pest management. A sustainable fungal treatment was developed to preserve fungal inoculum in potting soil and reduce thrips populations. Combining cooked, oven-dried millet with BotaniGard® (a commercial form of Beauveria bassiana strain GHA) to potting soil increased spore production and persistence of the fungus in the soil. In treated pots with millet, spore concentrations were 3–4 times greater after 30 days compared with spore yields at 10 days. The number of WFT adults was significantly lower in the marigold pots treated with GHA mix + millet than untreated controls, 12% and 10% in treated pots and 70% and 68% in untreated pots in sterile and non-sterile soil, respectively. Incorporation of millet in the potting mix enhanced the effect of the fungal treatments by providing a nutritive substrate on which the fungus could become established. This method is relatively inexpensive and easy for growers to use in greenhouses because granular formulations of B. bassiana are not commercially available.

Sexual interaction is an important activity that determines the reproductive schedule of organisms and can ultimately influence the fitness traits of both sexes. Although the influence of sexual interaction on the fitness of females has been extensively determined, little is known about the effects on males, which often have different mating strategies and optimal mating regimes from those of females. To understand how mating regimes (timing and frequency) modulate the fitness in both sexes, we used spider mite (Tetranychus urticae) to investigate the influence of delayed mating and repeated mating on the fitness of male and female. For females, the unmated and the delayed mating females outlived those mated immediately after adult emergence. The repeated mating shortened the lifespan of females that mated at 1-day-old, but not that mated 7-day-old. However, no significant variation in lifespan was observed for males across different mating regimes. We found although delayed mating significantly reduced the daily reproductive rate of the females, there was no significant difference in lifetime reproduction of females across treatments because the delayed mating females increased their reproductive lifespan as a compensation. Our study highlighted that the time and frequency of sexual interaction showed a sex-specific consequence on male and female spider mites, indicating that sexual interaction incurs a higher cost to females which have a much lower optimal mating frequency than males.

The alien cynipid wasp Dryocosmus kuriphilus Yasumatsu, 1951 is a serious pest of chestnuts (Castanea spp.) in Japan, North America and Europe, causing fruit losses while inducing galls in buds. While D. kuriphilus galls have a recognizable and roughly invariable globular shape, their size varies, reaching up to 4 cm in diameter. Among other factors, such variation may depend on different climatic conditions in different attacked areas. Here, we sampled and measured 375 D. kuriphilus galls from 25 localities throughout the Iberian Peninsula, including both cold and rainy northern (Eurosiberian) areas and warm and dry central-southern (Mediterranean) areas, to test the effects of climate and geographical location on gall morphology. The analyses indicate that gall mass and volume follow a pattern that can be associated with a climatic cline. In particular, the Eurosiberian galls were smaller than the Mediterranean galls according to differences in climatic conditions. In the southern areas, the greater insolation regime does not allow the chestnut trees to be distributed at lower altitudes, but the high rainfall and humidity regime of the mountain enclaves allow their presence. These conditions of insolation and precipitation seem to influence the morphological characteristics of the galls of D. kuriphilus.

A southern population (S) from Xiushui County (29°1′N, 114°4′E) and a northern population (N) from Shenyang city (41°48′N, 123°23′E) of the cabbage beetle, Colaphellus bowringi vary greatly in their life-history traits, and may serve as an excellent model with which to study the inheritance of life-history traits. In the present study, we performed intraspecific hybridization using the two populations, comparing the key life-history traits (fecundity, development time, body weight, growth rate, and sexual size dimorphism (SDD)) between the two populations (S♀ × S♂ and N♀ × N♂) and their two hybrid populations (S♀ × N♂ and N♀ × S♂ populations) at 19, 22, 25, and 28°C. Our results showed that there were significant differences in life-history traits between the two parental populations, with the S population having a significantly higher fecundity, shorter larval development time, larger body weight, higher growth rate, and greater weight loss during metamorphosis than the N population at almost all temperatures. However, these life-history traits in the two hybrid populations were intermediate between those of their parents. The life-history traits in the S × N and N × S populations more closely resembled those of the maternal S population and N population, respectively, showing maternal effects. Weight loss for both sexes was highest in the S population, followed by the S × N, N × S, and N populations at all temperatures, suggesting that larger pupae lost more weight during metamorphosis. The changes in SSD with temperature were similar between the S and the S × N populations and between the N and the N × S populations, also suggesting a maternal effect. Overall, our results showed no drastic effect of hybridization on C. bowringi, being neither negative (hybrid inferiority) nor positive (heterosis). Rather, the phenotypes of hybrids were intermediate between the phenotypes of their parents.

The vegetation community affects the composition and diversity of the insect community in grasslands. To explore the effects of vegetation management on insect community abundance and diversity, regular mowing of the vegetation was conducted, and tall fescue (Festuca arundinacea) and ryegrass (Lolium perenne) were exclusively planted at Tianjin Binhai International Airport. A total of 1886 insects were collected, representing 8 orders, 23 families, and 29 species; Acrididae (Orthoptera), Coccinellidae (Coleoptera), and Chironomidae (Diptera) were the dominant taxa. The abundance and biomass of insects in the turf areas were significantly lower than those in the control area and were reduced by 45.8 and 48.5% in the ryegrass area, respectively. In all areas, insect abundance and biomass peaked in summer, and the abundance of individuals and taxa decreased as the temperature decreased. Greater diversity and richness were found in summer compared with the other two seasons, and the turf areas had lower diversity and richness indices than the control areas in spring and summer. Our results suggest that the abundance, biomass and diversity of insects can be effectively decreased by artificial regulation of grassland vegetation at the airport, the planting of a single turfgrass – specifically ryegrass had the greatest effect. The present study provides a theoretical basis for the ecological control of insects at the airport.

Inbreeding costs can be high in haplodiploid hymenopterans due to their particular mechanism of sex determination (i.e., single-locus complementary sex-determination system, sl-CSD), as it can lead to the production of sterile males. Therefore, mechanisms contributing to reduced inbred matings can be beneficial. In this sense, asynchronous nest departure of sibling drones and gynes could reduce kin encounters in social hymenopterans. Using six observation colonies, we determined under field conditions the nest departure behaviour of sibling reproductives of the social wasp Vespula germanica (Hymenoptera: Vespidae). We determined that sexuals leave the nests definitively and detected asynchronous departure not fixed to a particular caste at a seasonal scale in some colonies, as gynes or drones delayed their departure as a function of the departure of the opposite sex, depending on the colony. At a higher temporal resolution (i.e., within a day), we discovered that drones consistently began to leave nests 1 h before gynes and this difference was driven by those individuals that left on the same day as did the opposite-sex kin. Even though other mechanisms such as polyandry and differential dispersal could also be important at reducing inbred matings in the species, the observed departure patterns (i.e., in some colonies actually leave together with the opposite caste, while in others temporal segregation seems to occur) from nests could be complementary to the former and be important at reducing the negative effects of inbreeding in this invasive species.

Dryocosmus kuriphilus (Hymenoptera: Cynipidae) is a global invasive gall wasp and a pest of chestnuts (Castanea spp.). A study of the Chalcidoidea parasitoid community of D. kuriphilus was undertaken over two years, from March 2017 to March 2019, at 15 sites in south and northwest Spain (Málaga and Galicia regions). More than 18,000 galls were collected, and 1153 parasitoids belonging to 22 species of seven chalcidoid families, plus two individuals of an inquiline Cynipidae, Synergus facialis, emerged. Richness was higher in the Málaga region, with 20 species, while 17 parasitoids and one inquiline were identified in Galicia. The parasitism rate of native chalcid parasitoid species in both regions was low. Eupelmus urozonus and Mesopolobus lichtensteini were the most abundant native species. Mesopolobus tibialis was a dominant species in south Spain, while Ormyrus pomaceus was a dominant species in northwest Spain. Our results revealed the existence of a sub-community of univoltine, probably host specialized, parasitoids in south Spain, which overwinter in galls, exhibiting a similar life cycle to Torymus sinensis. These species were Torymus notatus, Aulogymnus bicolor, Aulogymnus obscuripes and Aulogymnus balani. Data on the recovery of T. sinensis after release in the south Spain region show it to be well established, but its numbers are still low in northwest Spain.

The spotted wing drosophila, Drosophila suzukii, is an invasive pest in Europe and North America. Access to resources may be challenging in late fall, winter and early spring and flies may suffer from food deprivation along with cold stress in these periods. Whereas a plethora of studies have been performed on the overwintering capacity of D. suzukii, the effects of starvation on the fly's cold tolerance have not been addressed. In the present study, young D. suzukii adults (reared at 25°C, LD 12:12 h) were deprived of food for various periods (0, 12, 24 and 36 h), after which chill coma recovery time, critical thermal minimum, as well as acute and chronic cold tolerance were assessed. Additionally, the body composition of adults (body mass, water content, total lipid, glycerol, triglycerides, glucose and proteins) before and after starvation periods was analysed to confirm that starvation had detectable effects. Starved adults had a lower body mass, and both lipid and carbohydrate levels decreased with starvation time. Starvation slightly increased critical thermal minimum and affected chill coma recovery time; however, these changes were not gradual with starvation duration. Starvation promoted acute cold tolerance in both sexes. This effect appeared faster in males than in females. Food deprivation also led to enhanced survival to chronic cold stress. Short-term starvation was thus associated with significant changes in body composition in D. suzukii, and these alterations could alter some ecologically relevant traits related to cold tolerance, particularly in females. Our results suggest that food deprivation during short time (<36 h) can promote cold tolerance (especially survival after a cold stress) of D. suzukii flies. Future studies should address the ecological significance of these findings as short food deprivation may occur in the fields on many occasions and seasons.

Cytochrome P450 monooxygenases (P450s) play significant roles in protecting organisms from abiotic stress damage. Here, we report the sequence and characterization of a P450s gene (AccCYP4AV1), isolated from Apis cerana cerana Fabricius. The open reading frame of AccCYP4AV1 is 1506 base pairs long and encodes a predicted protein of 501 amino acids and 57.84 kDa, with an isoelectric point of 8.67. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis indicated that AccCYP4AV1 is more highly expressed in the midgut than in other tissues. In addition, the highest expression occurs in newly emerged adult workers, followed by the first instar of the larval stage. In addition, the expression of the AccCYP4AV1 was upregulated by low temperature (4 °C), ultraviolet radiation, hydrogen peroxide, paraquat, and dichlorvos treatments. In contrast, AccCYP4AV1 transcription was downregulated by other abiotic stress conditions: exposure to increased temperature (44 °C), deltamethrin, cadmium chloride, and mercury (II) chloride. Moreover, when AccCYP4AV1 was knocked-down by RNA interference, the results suggested that multiple antioxidant genes (AccsHSP22.6, AccSOD2, AccTpx1, and AccTpx4) were downregulated and antioxidant genes AccGSTO1 and AccTrx1 were upregulated. The activity levels of peroxidase and catalase were upregulated in the AccCYP4AV1-knocked-down samples, compared with those in the control groups. These findings suggest that the AccCYP4AV1 protein might be involved in the defense against abiotic stress damage.

The predatory mirid Macrolophus praeclarus is widely distributed throughout the Americas, and is reported to prey upon several horticultural pest species. However, little is known about its biology, thermal requirements, crop odour preferences, phytophagy, and capability to induce defensive responses in plants. When five temperatures studied (20, 25, 30, 33 and 35°C) were tested and Ephestia kuehniella was used as prey, the developmental time from egg to adult on tomato, was longest at 20°C (56.3 d) and shortest at 33°C (22.7 d). The ability of nymphs to develop to adults decreased as the temperature increased, with the highest number of nymphs reaching the adult stage at 20°C (78.0%) and lowest at 35°C (0%). The lower and upper developmental thresholds were estimated at 11.2° and 35.3°C, respectively. The maximum developmental rate occurred at 31.7°C and the thermal constant was 454.0 ± 8.1 degree days. The highest predation rate of E. kuehniella eggs was obtained at 30°C. In Y-tube olfactory choice tests, M. praeclarus selected tomato, sweet pepper and eggplant odours more frequently than no plant control treatment. Macrolophus praeclarus feeding did not damage tomato plants compared to another zoophytophagous mirid, Nesidiocoris tenuis, which caused necrotic rings. The phytophagy of M. praeclarus induced defensive responses in tomato plants through the upregulation of the jasmonic acid metabolic pathway. The implications of the findings for using M. praeclarus in tomato biological control programmes in the Americas are discussed.

Anastrepha fraterculus (Diptera: Tephritidae) is a major barrier to fruit production and exportation. In Brazil, the native parasitoid Aganaspis pelleranoi (Hymenoptera: Figitidae) and the exotic parasitoid Diachasmimorpha longicaudata (Hymenoptera, Braconidae) stand out as biological control agents. Knowledge of the factors that affect interactions among parasitoids, A. fraterculus, and host fruits may enhance the use of these agents in biological control programmes. This study evaluated the chemotaxis and parasitism of A. pelleranoi and D. longicaudata females reared on A. fraterculus larvae and kept on an artificial diet, red guava (Psidium guajava) or apple (Malus domestica). Females of both parasitoid species that emerged from larvae raised on artificial diet, guava or apple, were tested to Y olfactometer choice tests. In the parasitism tests, both parasitoid species were made to choose between A. fraterculus larvae brushed with water, apple pulp or guava pulp. D. longicaudata females from artificial diet (control) did not distinguish between fruit odours; however, females of D. longicaudata from larvae kept in apple or guava directed to the odours of their original fruit. The greatest parasitism for D. longicaudata occurred in the units that contained the pulp in which the larvae grew. A. pelleranoi from artificial diet preferred guava odours, including the females kept in apple. Similar results were observed in the parasitism bioassays. Our results found that A. fraterculus larval feeding influenced search behaviour and parasitism of D. longicaudata, whereas A. pelleranoi rearing experience did not affect its host choices.

The corn leafhopper Dalbulus maidis (Hemiptera: Cicadellidae), a specialist herbivore, is the cause of serious losses in maize yield for its capacity to transmit three important plant pathogens. They are also active phloem feeders, that insert stylets into the plant as they feed. Females place their eggs endophytically, totally inserted in the central midrib or the leaf blades, leaving conspicuous openings in the place where the ovipositor was inserted. In spite of the consequences that feeding and oviposition may have on the water status of the plant and the production of biomass, direct damage caused by the leafhopper has been only scarcely studied. In the present contribution, we measured biomass loss due to direct damage in maize plants under two watering regimes, with water supply ad libitum and with a watering restricted regime, emulating the most frequent field conditions. Moreover, we analyzed the effects of increasing densities of the vector on the biomass loss and plant mortality and the effects of females vs males. We observed that a density of 10 insects is sufficient to cause damage to 10-day-old seedlings, even in an ad libitum watering regime; however, in drought conditions, damage can be significantly greater, causing plant mortality. Also, females cause more damage than males, due to their oviposition habits.

Tetrastichus giffardianus is a gregarious koinobiont endoparasitoid of tephritids, including Ceratitis capitata, which is one of the most important fruit pests worldwide. The objective of this study was to evaluate the effect of constant temperatures on the biology and development of the egg-adult period of T. giffardianus in larvae/pupae of C. capitata to construct a fertility life table. The study was carried out in climatic chambers at constant temperatures of 15, 20, 25, 30, and 35 ± 1°C, 70 ± 10% relative humidity and a photophase of 12 h. Complete egg and larval development occurred only at temperatures of 20, 25, and 30°C. The mean longevity of males and females was inversely proportional to temperature. The time of development of the pre-imaginal period was influenced by temperature and ranged from 41 days at 20°C to 11 days at 25°C. The best T. giffardianus performance occurred at 25°C when the intrinsic rate of increase was 0.21, the mean generation time was 20.43 days, the time required to double the population was 3.33 days, and the finite rate of population increase (λ) was 1.23. The results of this study provided information for the establishment of mass rearing and an evaluation of the adaptability of T. giffardianus to different environments, which is necessary for the use of this parasitoid in the biological control of tephritid pests, particularly C. capitata.

Henosepilachna vigintioctopunctata is one of the most serious insect pests to a large number of nightshades and cucurbits. RNA interference (RNAi) triggered by double-stranded RNA (dsRNA) offers a reduced risk approach to control the beetle. Identification of amenable target genes and determination of appropriate life stage for dsRNA treatment are two critical steps in order to improve RNAi efficiency. In the present paper, we identified three vATPase genes, namely HvvATPaseC, HvvATPaseE and HvvATPaseH. We found that the three transcripts were widely expressed in the eggs, first- to fourth-instar larvae, prepupae, pupae and adults. They were abundantly transcribed in the hindgut and Malpighian tubules, in contrast to the epidermis and fat body. Three days' ingestion of dsvATPaseC, dsvATPaseE and dsvATPaseH by the fourth-instar larvae significantly decreased corresponding transcript level by 90.1, 88.9 and 97.2%, greatly reduced larval fresh weight by 28.0, 29.9 and 28.0%, and caused 66.7, 100 and 78.7% larval lethality respectively. Comparably, 3 days' exposure of the third-instar larvae to dsvATPaseC significantly reduced HvvATPaseC mRNA level by 89.5%, decreased approximately 80% of the larval fresh weight, and killed 100% of the treated larvae. Therefore, the three vATPase genes, especially HvvATPaseE, are potential amenable target genes and young larvae are more susceptible to dsRNA. Our findings will enable the development of the dsRNA-based pesticide to control H. vigintioctopunctata.

In Argentina, the mosquito Aedes aegypti (L.) (Diptera: Culicidae) is distributed from subtropical to temperate climates. Here, we hypothesized that the expansion of Ae. aegypti into colder regions is favoured by high-phenotypic plasticity and an adaptive inhibition of egg hatching at low temperatures. Thus, we investigated the hatching response of eggs of three populations: one from a subtropical region (Resistencia) and two from temperate regions (Buenos Aires City and San Bernardo) of Argentina. Eggs collected in the field were raised in three experimental colonies. F1 eggs were acclimated for 7 days prior to immersion at 7.6 or 22°C (control eggs). Five immersion temperatures were tested: 7.6, 10.3, 11.8, 14.1 and 16°C (range of mean winter temperatures of the three localities). A second immersion at 22°C was performed 2 weeks later to assess the inhibition to hatch under favourable conditions. After the first immersion, we compared the proportions of hatched eggs and dead larvae among treatment levels, whereas after the second immersion we compared the hatching response among the three populations. The factors that most influenced the egg hatching response were the geographical origin of the populations and the immersion temperature, but not the acclimation temperature. The proportions of hatching and larval mortality at low temperatures were higher for Resistencia than for Buenos Aires and San Bernardo, whereas the hatching response at ambient temperature was lower for San Bernardo than for Buenos Aires and Resistencia. The results support the hypothesis that populations from colder regions show an adaptive inhibition of egg hatching.