In different parts of the world, aphid populations and their natural enemies are influenced by landscapes and climate. In the Neotropical region, few long-term studies have been conducted, maintaining a gap for comprehension of the effect of meteorological variables on aphid population patterns and their parasitoids in field conditions. This study describes the general patterns of oscillation in cereal winged aphids and their parasitoids, selecting meteorological variables and evaluating their effects on these insects. Aphids exhibit two annual peaks, one in summer–fall transition and the other in winter-spring transition. For parasitoids, the highest annual peak takes place during winter and a second peak occurs in winter–spring transition. Temperature was the principal meteorological regulator of population fluctuation in winged aphids and parasitoids during the year. The favorable temperature range is not the same for aphids and parasitoids. For aphids, temperature increase resulted in population growth, with maximum positive effect at 25°C. Temperature also positively influenced parasitoid populations, but the growth was asymptotic around 20°C. Although rainfall showed no regulatory function on aphid seasonality, it influenced the final number of insects over the year. The response of aphids and parasitoids to temperature has implications for trophic compatibility and regulation of their populations. Such functions should be taken into account in predictive models.

The coffee white stem borer, Xylotrechus quadripes Chevrolat (Coleoptera: Cerambycidae), is a major destructive pest of Coffea arabica L. (Gentianales: Rubiaceae), widely planted in many Asian countries, including China. Quantitative real-time polymerase chain reaction (qRT-PCR) is a common method for quantitative analysis of gene transcription levels. To obtain accurate and reliable qRT-PCR results, it is necessary to select suitable reference genes to different experimental conditions for normalizing the target gene expression. However, the stability of the expression of reference genes in X. quadripes has rarely been studied. In this study, the expression stability of nine candidate reference genes were investigated under biotic and abiotic conditions for use in qRT-PCR's normalization. By integrating the results of four algorithms of NormFinder, BestKeeper, geNorm, and RefFinder, the optimal reference gene combinations in different experimental conditions were performed as follows: RPL10a and EIF3D were the optimal reference genes for developmental stage samples, EIF4E, RPL10a, and RPS27a for tissue samples, V-ATP and EF1α for the sex samples, EIF3D and V-ATP for temperature treatment, RPS27a and RPL10a for insecticide stress, and RPL10a, RPS27a, and EF1α for all the samples. This study will help to obtain the stable internal reference genes under biotic and abiotic conditions and lay the foundation for in-depth functional research of target genes or genomics on olfactory molecular mechanisms, temperature adaptability, and insecticide resistance in X. quadripes.

Natural enemies that impact pest populations must be understood in order to build integrated pest control strategies and to understand the most important aspects affecting pest dynamics. Haloxylon ammodendron (C. A. Mey.) Bunge is an important perennial plant species extensively used in sand stabilization and wind prevention in arid areas. This study aimed to determine the main defoliators that damage H. ammodendron and the parasitoid complex associated with them. Twelve species of defoliators were found in Northern Xinjiang, and Teia dubia (Tauscher) (Lepidoptera: Lymantriidae), Scrobipalpa sp. (Lepidoptera: Gelechiidae), and Eucharia festiva Hüfnagel (Lepidoptera: Arctiidae) were the dominant pests. T. dubia is the predominant defoliator with three generations a year. Northwest China, Central Asia, and the Mediterranean region are potentially suitable habitats for T. dubia in the world, while Xinjiang is the primary distribution area in China. Parasitoids belonging to seven species and four families were reared from the larvae of T. dubia, they were all endoparasitoids and koinobiont. Cotesia sp. (Hymenoptera: Braconidae) is the dominant parasitoid and prefer to parasitic in the 3rd–5th instar larvae. The present study provides the basis for understanding the species composition and natural enemies of lepidopteran defoliators. It will be an effective tool for the integrated pest management programs of H. ammodendron forest.

Cytochrome P450 monooxygenases play a key role in pest resistance to insecticides by detoxification. Four new P450 genes, CYP6AS160, CYP6AS161, CYP4AB73 and CYP4G232 were identified from Solenopsis invicta. CYP6AS160 was highly expressed in the abdomen and its expression could be induced significantly with exposure to fipronil, whereas CYP4AB73 was not highly expressed in the abdomen and its expression could not be significantly induced following exposure to fipronil. Expression levels of CYP6AS160 and CYP4AB73 in workers were significantly higher than that in queens. RNA interference-mediated gene silencing by feeding on double-stranded RNA (dsRNA) found that the levels of this transcript decreased (by maximum to 64.6%) when they fed on CYP6AS160-specific dsRNA. Workers fed dsCYP6AS160 had significantly higher mortality after 24 h of exposure to fipronil compared to controls. Workers fed dsCYP6AS160 had reduced total P450 activity of microsomal preparations toward model substrates p-nitroanisole. However, the knockdown of a non-overexpressed P450 gene, CYP4AB73 did not lead to an increase of mortality or a decrease of total P450 activity. The knockdown effects of CYP6AS160 on worker susceptibility to fipronil, combined with our other findings, indicate that CYP6AS160 is responsible for detoxification of fipronil. Feeding insects dsRNA may be a general strategy to trigger RNA interference and may find applications in entomological research and in the control of insect pests in the field.

The Mormon cricket (MC), Anabrus simplex Haldeman, 1852 (Orthoptera: Tettigoniidae), has a long and negative history with agriculture in Utah and other western states of the USA. Most A. simplex populations migrate in large groups, and their feeding can cause significant damage to forage plants and cultivated crops. Chemical pesticides are often applied, but some settings (e.g. habitats of threatened and endangered species) call for non-chemical control measures. Studies in Africa, South America, and Australia have assessed certain isolates of Metarhizium acridum as very promising pathogens for Orthoptera: Acrididae (locust) biocontrol. In the current study, two isolates of Metarhizium robertsii, one isolate of Metarhizium brunneum, one isolate of Metarhizium guizhouense, and three isolates of M. acridum were tested for infectivity to MC nymphs and adults of either sex. Based on the speed of mortality, M. robertsii (ARSEF 23 and ARSEF 2575) and M. brunneum (ARSEF 7711) were the most virulent to instars 2 to 5 MC nymphs. M. guizhouense (ARSEF 7847) from Arizona was intermediate and the M. acridum isolates (ARSEF 324, 3341, and 3609) were the slowest killers. ARSEF 2575 was also the most virulent to instar 6 and 7 nymphs and adults of MC. All of the isolates at the conidial concentration of 1 × 107 conidia ml−1 induced approximately 100% mortality by 6 days post application of fungal conidia. In conclusion, isolates ARSEF 23, ARSEF 2575, and ARSEF 7711 acted most rapidly to kill MC under laboratory conditions. The M. acridum isolates, however, have much higher tolerance to heat and UV-B radiation, which may be critical to their successful use in field application.

Diadegma semiclausum is an important parasitoid wasp and widely used in the biological control of the diamondback moth, Plutella xylostella, one of the most destructive pests of cruciferous plants. Insect olfactory system is critical in guiding behaviors including feeding, mating, and oviposition, in which odorant binding proteins (OBPs) and odorant receptors (ORs) are two key components. However, limited attention has been paid to D. semiclausum olfactory genes. In this study, a transcriptome sequencing was performed on the RNA samples extracted from D. semiclausum male and female adult antennae. A total of 17 putative OBP and 67 OR genes were annotated and further compared to OBPs and ORs from P. xylostella, and other hemipteran parasitoid species. The expression patterns of D. semiclausum OBPs between male and female antennae were examined using reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR. Six OBPs (DsemOBP 6, 7, 8, 9, 10, and 14) demonstrated significantly higher expression levels in females than in males, which may assist in female D. semiclausum host-seeking and oviposition behaviors. This study advances our understanding of the olfactory system of D. semiclausum at the molecular level and paves the way for future functional studies aiming at increasing the efficacy to control P. xylostella by using D. semiclausum.

Secondary metabolites are central to understanding the evolution of plant–animal interactions. Direct effects on phytophagous animals are well-known, but how secondary consumers adjust their behavioural and physiological responses to the herbivore's diet remains more scarcely explored for some metabolites. Caffeine is a neuroactive compound that affects both the behaviour and physiology of several animal species, from humans to insects. It is an alkaloid present in nectar, leaves and even sap of numerous species of plants where it plays a role in chemical defences against herbivores and pathogens. Caffeine effects have been overlooked in generalist herbivores that are not specialized in coffee or tea plants. Using a host–parasitoid system, we show that caffeine intake at a relatively low dose affects longevity and fecundity of the primary consumer, but also indirectly of the secondary one, suggesting that this alkaloid and/or its effects can be transmitted through trophic levels and persist in the food chain. Parasitism success was lowered by ≈16% on hosts fed with caffeine, and parasitoids of the next generation that have developed in hosts fed on caffeine showed a reduced longevity, but no differences in mass and size were found. This study helps at better understanding how plant secondary metabolites, such as caffeine involved in plant–animal interactions, could affect primary consumers, could have knock-on effects on upper trophic levels over generations, and could modify interspecific interactions in multitrophic systems.

Two Polygonaceae weeds, Rumex dentatus L. and Polygonum glabrum Willd. are abundant in wheat- and rice-fields, respectively, in India. Galerucella placida Baly (Coleoptera: Chrysomelidae) is a biocontrol agent of these two weeds. The importance of long-chain alkanes and free fatty acids present in leaf surface waxes of these weeds was assessed as short-range attractant and ovipositional stimulant in G. placida females. Extraction, TLC, GC-MS and GC-FID analyses demonstrated 19 n-alkanes from n-C14 to n-C35 and 14 free fatty acids from C12:0 to C22:0 in leaf surface waxes. Hentriacontane was predominant among alkanes in both weeds, while oleic acid and docosanoic acid were predominant among free fatty acids in R. dentatus and P. glabrum, respectively. Females of G. placida were attracted toward one leaf equivalent surface wax of both weeds against the control solvent (petroleum ether) in a short Y-tube olfactometer bioassay. But, the insect could not differentiate between one leaf equivalent surface wax of R. dentatus and P. glabrum, indicating that both weed leaves were equally attractive in females. A synthetic blend of either 2.44, 35.57 and 23.58 μg ml−1 of octadecane, heptacosane and nonacosane, respectively, resembling the amounts present in one leaf equivalent surface wax of R. dentatus or 4.08, 19.54 and 23.58 μg ml−1 of octadecane, palmitoleic acid and docosanoic acid, respectively, resembling the amounts present in one leaf equivalent surface wax of P. glabrum acted as short-range attractant and ovipositional stimulant in G. placida. These results could be a basis for host plant specificity of the biocontrol agent.

Embryo development in eggs of the spittlebug Mahanarva spectabilis (Distant) (Hemiptera: Cercopidae) passes through four phases (known as S1 to S4) being stopped at S2 during diapause. Studies about the molecular basis of diapause in spittlebugs are nonexistent. Here, we analyzed proteins from non-diapausing (ND), diapausing (D) and post-diapausing (PD) eggs of the spittlebug M. spectabilis. In total, we identified 87 proteins where 12 were in common among the developmental and diapause phases and 19 remained as uncharacterized. Non-diapausing eggs (S2ND and S4ND) showed more proteins involved in information storage and processing than the diapausing ones (S2D). Eggs in post-diapausing (S4PD) had a higher number of proteins associated with metabolism than S2D. The network of protein interactions and metabolic processes allowed the identification of different sets of molecular interactions for each developmental and diapause phases. Two heat shock proteins (Hsp65 and Hsp70) along with two proteins associated with intracellular signaling (MAP4K and a serine/threonine-protein phosphatase) were found only in diapausing and/or post-diapausing eggs and are interesting targets to be explored in future experiments. These results shine a light on one key biological process for spittlebug survival and represent the first search for proteins linked to diapause in this important group of insects.

The reproductive traits of the mymarid wasp Cosmocomoidea annulicornis (Ogloblin) (Hymenoptera: Mymaridae) attacking eggs of the sharpshooter Tapajosa rubromarginata (Signoret) (Hemiptera: Cicadellidae) were evaluated under laboratory conditions. Bioassays were carried out to estimate the realized fecundity and egg load of females. The ovigeny index was calculated and different biological traits, such as body size, oöcyte length, gaster length and wing length, were analysed to assess significant associations between these traits and the species fitness. In addition, the effect of host availability and feeding on longevity and potential fecundity throughout life and the effect of female age on egg maturation dynamics were assessed. The results showed that C. annulicornis is a strongly synovigenic species. A positive correlation was found between fecundity and longevity of the females and between body size and oöcyte length. Contrary to expected, body size was not related to fecundity and longevity. Females lived significantly longer in the presence of hosts and honey than when they were host-deprived and honey-fed or both host and honey deprived. Host availability had a significant effect on the amount of eggs laid by C. annulicornis females. Female age was negatively associated with oöcyte length. Furthermore, females were able to mature additional eggs as they aged, nevertheless, when host-deprived, senescent females presented significantly less mature eggs than younger ones, suggesting a possible egg oosorption. These results might contribute to a better understanding of the reproductive potential of this species as a biocontrol agent.

Invasive Tephritid fruit flies are a global threat to both agriculture and horticulture industries. Biosecurity has played a critical role in reducing their damage but becomes more and more challenging after several key chemical pesticides were banned or withdrawn for health or environmental reasons. This has led to non-chemical approaches including heat and cold treatments being broadly utilized to get rid of fruit fly infestation. However, the molecular mechanisms to kill the flies underlying these stressors are not clear yet. This knowledge will certainly help refine current post-harvest treatment strategies and develop more efficient, cost-effective and environmentally friendly approaches for fruit fly management. Previously, the molecular response of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) to heat was examined thoroughly, in which 31 key genes were identified with significant changes in expression levels and their high-resolution expression timeline was constructed across 11 timepoints. However, whether these candidate genes respond to cold in the same way was unknown. Here, a temperature bioassay was conducted and the expression profiles of these genes were investigated across the same 11 timepoints using cold treatment. The results showed that most of candidate genes exhibited divergent expression profiles compared to heat treatment, suggesting that the fly molecular response to cold may be different from those to heat. This study provides new knowledge of Tephritid fruit fly response to cold at a molecular level, which could aid in improving current fruit fly management and facilitate the development of new strategies to control this serious horticultural insect pest.

To explore the characteristics of the mitochondrial genome (mitogenome) of the squeaking silkmoths Rhodinia, a genus of wild silkmoths in the family Saturniidae of Lepidoptera, and reveal phylogenetic relationships, the mitogenome of Rhodinia fugax Butler was determined. This wild silkmoth spins a green cocoon that has potential significance in sericulture, and exhibits a unique feature that its larvae can squeak loudly when touched. The mitogenome of R. fugax is a circular molecule of 15,334 bp long and comprises 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and an A + T-rich region, consistent with previous observations of Saturniidae species. The 370-bp A + T-rich region of R. fugax contains no tandem repeat elements and harbors several features common to the Bombycidea insects, but microsatellite AT repeat sequence preceded by the ATTTA motif is not present. Mitogenome-based phylogenetic analysis shows that R. fugax belongs to Attacini, instead of Saturniini. This study presents the first mitogenome for Rhodinia genus.

Knowledge on reproductive strategies and host use in insect parasitoids is fundamental for biological control purposes. Superparasitism and multiparasitism, oviposition in a previously parasitized host by a female of the same or different species, respectively, may impact pest management decisions. Our objective was to determine the occurrence of superparasitism and multiparasitism in three species of native larval−pupal solitary endoparasitoids that attack Anastrepha Shiner species (Diptera: Tephritidae) in the Neotropical region, and the possible effect on offspring fitness parameters. Doryctobracon crawfordi (Viereck), Utetes anastrephae (Viereck), and Opius hirtus (Fischer) occur in sympatry in Mexico, and are currently under consideration for use as biocontrol agents. Experiments were conducted under laboratory conditions with females acting alone (self-superparasitism), females in groups of the same species (conspecific superparasitism), and females in mixed groups (multiparasitism). Our results showed that self-superparasitism is an uncommon strategy in the three native species and is rare under conditions of intraspecific competition. In the case of multiparasitism, a higher number of immature stages of U. anastrephae was observed, compared to those of D. crawfordi and O. hirtus. However, it is not clear yet if this was due to some adult female trait or to the competitive ability of the larvae. We conclude that most females of the native species studied appeared to avoid superparasitism, specifically when acting alone, suggesting a high discrimination ability, which is probably a result of a close relationship and evolutionary history with Anastrepha hosts.

Honeybees cannot synthesize arachidonic acid (ARA) themselves, only obtain it from food. Most pollen is deficient or contains a small amount of ARA. The necessity of supplementary ARA in bees’ diet has not been studied. The objective of this study was to investigate the effects of dietary ARA levels on the growth and immunity of Apis mellifera ligustica. A total of 25 honeybee colonies were randomly assigned to five dietary groups which were fed basic diets supplemented with 0, 2, 4, 6, and 8% of ARA. The diet with 4% ARA improved the body weight of newly emerged worker bees compared with the control group. Supplement of ARA in honeybee diets changed the fatty acid composition of honeybee body. SFA and MUFA contents of bees’ body declined, and PUFA content rised in the ARA group. Compared with the control group, the supplement of ARA in honeybee diets increased the contents of ARA, C22:6n-3 (DHA) and C18:3n-6 in bees’ body significantly, but decreased the contents of C16:1 and C18:3n-3. The diet supplied with 4% ARA reduced the mortality rate of honeybee infected with Escherichia coli. The activity of immune enzymes (phenoloxidase, antitrypsin, and lysozyme) and the mRNA expression levels of immune genes (defensin-2, toll, myd88, and dorsal) were improved by ARA diets to varying degrees depending on the ARA levels, especially 4% ARA. These results suggested that dietary ARA could improve the growth, survival, and immune functions of honeybees. Supplement of ARA in bees’ diet would be valuable for the fitness of honeybees.

Fall armyworm, Spodoptera frugiperda (J. E. Smith) is a polyphagous and highly destructive invasive insect pest of many crops. It was recently introduced into India and widely reported in almost all parts of India. Development of a temperature-based phenology model for predicting its rate of development and distribution will help in understanding the establishment and further spread of introduced invasive insect pests. Development, survival and reproduction parameters of S. frugiperda at six constant temperature conditions (15, 20, 25, 27, 30 and 35°C) were investigated and further validated with data generated under fluctuating temperature conditions. The estimated lower developmental threshold temperatures were 12.1°C for eggs, 11°C for larvae, 12.2°C for pupae, 15.13°C for males and 12.66°C for females. Degree-day (DD) requirements for the development of the different stages of S. frugiperda were 50, 250 and 200 DD for egg, larva and pupa, respectively. The best-fitted functions were compiled for each life stage to yield a phenology model, which was stochastically simulated to estimate the life table parameters. The developed phenology model predicted temperature ranges between 27 and 30°C as favourable for S. frugiperda development, survival and reproduction. The results revealed that maximum net reproductive rate (215.66 females/female/generation) and total fecundity (981.08 individuals/female/generation) were attained at 30°C constant temperature. The mean length of generations decreased from 74.29 days at 15°C to 38.74 days at 30°C. The maximum intrinsic rate of increase (0.138 females/female/day) and shortest doubling time (4.9 days) were also observed at 30°C. Results of simulated life table parameters showed high temperature-dependent development of S. frugiperda and complete development within all the tested constant temperature ranges (15–35°C). Simulated life table parameters for predicting risk indices of S. frugiperda in India indicated a significant increase in activity indices and establishment risk indices with a higher number of generations during future (2050 and 2070) climatic change scenarios compared to present conditions. Our results indicate that India will be highly suitable for the establishment and survival of S. frugiperda in future time periods.