Rhynchophorus ferrugineus, known as the Red Palm Weevil (RPW), is reported as a pest of up to 40 palm species. However, the susceptibility degree and the defense mechanisms of these species against this weevil are still poorly known. In Europe, the RPW is a major pest of Phoenix canariensis while other palm species, including the congeneric Phoenix dactylifera, seem to be less suitable hosts for this insect. The aim of our study was to compare the defensive response of P. dactylifera, Chamaerops humilis and Washingtonia filifera against R. ferrugineus and try to define the mechanisms of resistance that characterize these species. Bioassays were carried out to evaluate the mortality induced on RPW larvae by extracts from the leaf rachis of the studied palm species. Tests at semi-field scale were also conducted, based either on forced palm infestation, with larvae of RPW, or on natural infestation, with adult females. Rachis extracts from C. humilis and W. filifera caused 100% larval mortality after 2 days of exposure, while extracts of P. dactylifera did not impair larval survival. Independently of the effect of the leaf extracts, the weevils were unable to naturally infest the three palm species, although larval survival was high after forced infestation of the plants. We concluded that the observed lack of infestation of P. dactylifera by RPW is due to factors other than antibiosis. In W. filifera and C. humilis, although the presence of antixenosis mechanisms cannot be excluded, resistance to R. ferrugineus seems to rely on the presence of antibiosis compounds.

The effect of cover plants on arthropod functional biodiversity was investigated in a vineyard in Northern Italy, through a 3-year field experiment. The following six ground cover plants were tested: Sweet Alyssum; Phacelia; Buckwheat; Faba Bean; Vetch and Oat; control. Arthropods were sampled using different techniques, including collection of leaves, vacuum sampling and sweeping net. Ground cover plant management significantly affected arthropod fauna, including beneficial groups providing ecosystem services like biological control against pests. Many beneficial groups were attracted by ground cover treatments in comparison with control, showing an aggregative numerical response in the plots managed with some of the selected plant species. Alyssum, Buckwheat and ‘Vetch and Oat’ mixture showed attractiveness on some Hymenoptera parasitoid families, which represented 72.3% of the insects collected by sweeping net and 45.7 by vacuum sampling. Phytoseiidae mites showed a significant increase on leaves of the vineyard plots managed with ground covers, in comparison with control, although they did not show any difference among the treatments. In general, the tested ground cover treatments did not increase dangerous Homoptera populations in comparison with control, with the exception of Alyssum. The potential of ground cover plant management in Italian vineyards is discussed: the overall lack of potential negative effects of the plants tested, combined with an aggregative numerical response for many beneficials, seems to show a potential for their use in Northern Italy vineyards.

The southern harvester termite, Microhodotermes viator, is ecologically important due to its nutrient cycling activities and trophic interactions. Additionally, M. viator appears to have very long-lived colonies, which amplifies their effect on the environment. In order to estimate the longevity of a colony it is necessary to understand colony genetic structure. However, intra- and intercolonial genetic structure and levels of relatedness have not yet been examined in this species, likely due to a lack of microsatellite markers that effectively hybridize in this species. Here we describe the identification and characterization of seven microsatellite loci for M. viator, using an enriched approach and a preliminary test of their suitability for studies of fine-scale population genetic structure. Seven polymorphic loci were identified, none of which deviated from Hardy–Weinberg equilibrium. The loci had an average of 5.8 alleles per locus (range: 2–14) and an overall mean heterozygosity of 0.51 ± 0.3. Across all loci, population level pairwise FST values showed significant genetic differentiation. The loci described and preliminary genetic data presented here provide an invaluable tool for future studies of population structure and longevity in M. viator colonies.

The sexual performance of Anastrepha ludens males of the Tapachula-7 genetic sexing strain, produced via selection based on mating success, was compared with that of males produced without selection in competition with wild males. Mating competition, development time, survival, mass-rearing quality parameters and pheromone production were compared. The results showed that selection based on mating competitiveness significantly improved the sexual performance of offspring. Development time, survival of larvae, pupae and adults, and weights of larvae and pupae increased with each selection cycle. Differences in the relative quantity of the pheromone compounds (Z)-3-nonenol and anastrephin were observed when comparing the parental males with the F4 and wild males. The implications of this colony management method on the sterile insect technique are discussed.

Chitinases have an indispensable function in chitin metabolism and are well characterized in numerous insect species. Although the diamondback moth (DBM) Plutella xylostella, which has a high reproductive potential, short generation time, and characteristic adaptation to adverse environments, has become one of the most serious pests of cruciferous plants worldwide, the information on the chitinases of the moth is presently limited. In the present study, using degenerated polymerase chain reaction (PCR) and rapid amplification of cDNA ends-PCR strategies, four chitinase genes of P. xylostella were cloned, and an exhaustive search was conducted for chitinase-like sequences from the P. xylostella genome and transcriptomic database. Based on the domain analysis of the deduced amino acid sequences and the phylogenetic analysis of the catalytic domain sequences, we identified 15 chitinase genes from P. xylostella. Two of the gut-specific chitinases did not cluster with any of the known phylogenetic groups of chitinases and might be in a new group of the chitinase family. Moreover, in our study, group VIII chitinase was not identified. The structures, classifications and expression patterns of the chitinases of P. xylostella were further delineated, and with this information, further investigations on the functions of chitinase genes in DBM could be facilitated.

Developmental rates are highly variable, both within and between genotypes and populations. But the rationale for two differential (slow and fast) developmental rates within same cohort under varying prey supply has yet not been explored. For this purpose, we investigated the effect of scarce and abundant prey supply on slow and fast development at 27°C in two aphidophagous ladybirds, Menochilus sexmaculatus (Fabricius) and Propylea dissecta (Mulsant) and its effect on their body mass and reproductive attributes. The ladybirds were provided with scarce and abundant supply of Aphis craccivora Koch under standardized abiotic conditions in the laboratory. A clear bimodal (two peaks, where the first peak represented the fast developing individuals and the second peak slow developing individuals) pattern of distribution for both prey supplies was obtained, which got skewed with change in prey supply. On abundant prey supply, more fast developing individuals (139 M. sexmaculatus and 123 P. dissecta) were found and less (46 M. sexmaculatus and 36 P. dissecta) on scarce prey supply. Slow developing individuals had female biased sex ratio, higher longevity and lower body mass. Fast developing females laid higher number of eggs with higher egg viability. Results of the study are indicative of occurrence and constancy of the slow and fast developing individuals in the egg batch.

Phenol, also known as carbolic acid or phenic acid, is a priority pollutant in aquatic ecosystems. The present study has investigated metabolic activities and transcription profiles of cytochrome P450 enzymes in Chironomus kiinensis under phenol stress. Exposure of C. kiinensis larvae to three sublethal doses of phenol (1, 10 and 100 µM) inhibited cytochrome P450 enzyme activity during the 96 h exposure period. The P450 activity measured after the 24 h exposure to phenol stress could be used to assess the level (low or high) of phenol contamination in the environment. To investigate the potential of cytochrome P450 genes as molecular biomarkers to monitor phenol contamination, the cDNA of ten CYP6 genes from the transcriptome of C. kiinensis were identified and sequenced. The open reading frames of the CYP6 genes ranged from 1266 to 1587 bp, encoding deduced polypeptides composed of between 421 and 528 amino acids, with predicted molecular masses from 49.01 to 61.94 kDa and isoelectric points (PI) from 6.01 to 8.89. Among the CYP6 genes, the mRNA expression levels of the CYP6EW3, CYP6EV9, CYP6FV1 and CYP6FV2 genes significantly altered in response to phenol exposure; therefore, these genes could potentially serve as biomarkers in the environment. This study shows that P450 activity combined with one or multiple CYP6 genes could be used to monitor phenol pollution.

Diamondback moth or DBM is the major pest of Brassica vegetable production worldwide. Control has relied on insecticides, and DBM resistance to these compounds has evolved rapidly. We review and summarize data on DBM population dynamics across a large latitudinal gradient from southwest to northeast China: DBM is, on average, more common in southern locations than in northern locations. The species' phenology is consistent: in southern and central locations there is a decline during hot summer months, while in the north, the species can only exist in the summer following migrations from the south. A cohort-based discrete-time model, driven by daily maximum and minimum temperatures and rainfall, which was built using the DYMEX modelling software, captures the age-structured population dynamics of DBM at representative locations, with year round cropping and threshold-based insecticide applications. The scale of the simulated pest problem varies with cropping practices. Local production breaks and strict post-harvest crop hygiene are associated with lower DBM populations. Biological control appears to improve the management of DBM. Of the management strategies explored, non-threshold based applications of insecticides with reduced spray efficacy (due to poor application or resistance) appear the least effective. The model simulates the phenology and abundance patterns in the population dynamics across the climatic gradient in China reasonably well. With planned improvements, and backed by a system of field sampling and weather inputs, it should serve well as a platform for a local pest forecast system, spanning the range of DBM in China, and perhaps elsewhere.

Pheromone-binding proteins (PBPs) are believed to be involved in the recognition of semiochemicals. In the present study, western blot analysis, fluorescence-binding characteristics and immunolocalization of CmedPBP4 from the rice leaffolder, Cnaphalocrocis medinalis, were investigated. Western blot analysis revealed that CmedPBP4 showed obvious antenna-specific expression patterns in female and male antenna, and made a clearly different sex-biased expression. Immunocytochemical labeling revealed that CmedPBP4 showed specific expression in the trichoid sensilla. Competitive fluorescence binding assays indicated that CmedPBP4 could selectively recognize three sex pheromone components (Z13-18:Ac, Z11-16:Al and Z13-18:OH) and eleven rice plant volatiles, including cyclohexanol, nerolidol, cedrol, dodecanal, ionone, (−)-α-cedrene, (Z)-farnesene, β-myrcene, R-(+)-limonene, (−)-limonene, and (+)-3-carene. Meanwhile the CmedPBP4 detection of sex pheromones and host odorants was pH-dependent. Our results, for the first time, provide further evidence that trichoid sensilla might be play an important role in detecting sex pheromones and host plant volatiles in the C. medinalis moth. Our systematic studies provided further detailed evidence for the function of trichoid sensilla in insect semiochemical perception.

The Asian long-horned beetle (ALB), Anoplophora glabripennis (Motschulsky), is a highly polyphagous invasive pest with a broad range of host species, but showing relevant differences between infestation areas. Host preference and host colonization (female fecundity, egg and larval survival) were assessed in a population in Northern Italy by choice and no-choice experiments conducted in both field and laboratory conditions. During 5 years of field observations, ALB was found to infest seven genera of trees: Acer, Aesculus, Betula, Populus, Prunus, Salix and Ulmus. However, Acer, Betula, Ulmus and Salix resulted to be the preferred hosts corresponding to 97.5% (1112) of the 1140 infested trees. In both laboratory and field trials carried out on these four host genera, no-choice experiments recorded the highest host colonization of A. glabripennis on Acer trees, with the highest number of laid eggs and the lowest egg and larval mortality. Ulmus and Salix showed a lower number of laid eggs during laboratory choice test, but egg and larval mortality had mean values similar to Acer. On the contrary, despite the high number of Betula trees felled during the eradication plan carried out in the infestation area, this tree species showed the lowest beetle suitability in terms of number of laid eggs and insect survival. An overestimation of the number of infested Betula occurring during the tree survey may explain the discordance between high number of infested Betula and low beetle suitability. Instead, the large number of infested Acer recorded in the field was probably due to the high abundance of these trees occurring in parks and gardens within the infestation area and to the low adult dispersal of A. glabripennis. Overall, results from this study confirm that host species affects both beetle colonization and breeding performance. The study shows ALB host preference and host suitability varying between tree species, suggesting an ALB acceptance even of sub-optimal hosts.

For parasitoids, the host represents the sole source of nutrients for the developing immature. Subsequently, host quality is an important factor affecting immature development and the resulting fitness of the emerging parasitoid, with impacts on fecundity, longevity and offspring sex ratio. Host age is an integral component of host quality and a key factor in host selection by the female parasitoid. The current study aimed to investigate the effect of decreasing host quality (determined by increasing host age) on adult life history traits (size, wing loading, longevity, and fecundity) and nutritional reserves (protein, lipid and glycogen concentrations) of the parasitoid Trichogramma brassicae. Higher quality hosts resulted in the production of larger offspring with increased resource reserves and enhanced mobility. One-day-old eggs contained significantly more protein and triglyceride than 25- and 45–day-old eggs. Quality of host and fitness of reared wasps decreased due to host aging. Parasitoids reared on 1-day-old hosts were larger, with greater fecundity and longevity, a reduced wind loading index, and produced a higher proportion of female offspring when compared with those reared on 25- and 45-day-old hosts. In addition, wasps reared on 1-day-old hosts contained higher energy resources, as determined by triglyceride, glycogen and protein reserves, which are essential to successful offspring production. One-day-old hosts can therefore be considered as the best age for producing wasps with greater fitness, since they contain the highest amount of protein, glycogen, and triglyceride. This has implications for the mass rearing of T. brassicae and enhancing the efficacy of this biological control agent.

The sugarcane stem weevil, Acrotomopus atropunctellus (Boheman) (Curculionidae: Molytinae: Cholini) is an important economic pest from the Northwestern region of Argentina. Analyses of the headspace volatiles produced by separated males and females revealed one male-specific compound. Its structural identification is reported here in using gas chromatography coupled with mass spectroscopy analysis and chemical micro-reactions. Besides, two laboratory olfactometry assays allowed us to propose 6-methyl-5-hepten-2-one (sulcatone) as an aggregation pheromone for this insect, being attractive to both conspecific males and females. This compound is reported for the first time as involved in the Curculionidae family communication.

For optimal application of Nesidiocoris tenuis as a biological control agent, adequate field management and programmed mass rearing are essential. Mathematical models are useful tools for predicting the temperature-dependent developmental rate of the predator. In this study, the linear model and nonlinear models Logan type III, Lactin and Brière were estimated at constant temperatures and validated at alternating temperatures and under field conditions. N. tenuis achieved complete development from egg to adult at constant temperatures between 15 and 35°C with high survivorship (>80%) in the range 18–32°C. The total developmental time decreased from a maximum at 15°C (76.74 d) to a minimum at 33°C (12.67 d) and after that, increased to 35°C (13.98 d). Linear and nonlinear developmental models all had high accuracy (Ra2 >0.86). The maximum developmental rate was obtained between 31.9°C (Logan type III and Brière model for N1) and 35.6°C (for the egg stage in the Brière model). Optimal survival and the highest developmental rate fell within the range 27–30°C. The field validation revealed that the Logan type III and Lactin models offered the best predictions (95.0 and 94.5%, respectively). The data obtained on developmental time and mortality at different temperatures are useful for mass rearing this predator, and the developmental models are valuable for using N. tenuis as a biological control agent.

The robber fly Mallophora ruficauda is one of the most important apicultural pests in the Pampas region of Argentina. This species is a parasitoid of scarab beetle larvae. Females lay eggs away from the host, and the larvae perform active search behaviour toward Cyclocephala signaticollis third instar larvae, parasitoid's preferred host. This behaviour is mediated by host-related chemical cues produced in hosts’ fermentation chamber. Also, C. signaticollis larvae are attracted to fermentation chamber extracts. As scarab larvae have microbe-rich fermentation chamber, it has been suggested that microorganisms could be involved in the production of these semiochemicals. The aims of this work were first to ascertain the presence of microorganisms in the fermentation chamber of C. signaticollis larvae and second to determine the role of microorganisms in the orientation response of parasitoid and host larvae. We found that microorganisms-free C. signaticollis larvae showed deterioration in their development and did not produce the attractive semiochemicals. Therefore, we isolated fermentation chamber microorganisms of host larvae by means of different cultures media, and then, assayed different microorganisms’ stimuli by binary choice tests. We were able to isolate microorganisms and determine that M. ruficauda larvae are attracted to semiochemicals from protein degradation in the fermentation chamber. However, C. signaticollis larvae were not attracted to any semiochemicals associated with microorganisms’ activity in the fermentation chamber. Although we were unable to elucidate the exact role of gut microorganisms in host behaviour, we discuss their relevance in parasitoid host-seeking behaviour and host conspecific interaction in M. ruficauda–C. signaticollis system.

Insect defensins, are cationic peptides that play an important role in immunity against microbial infection. In the present study, an anionic defensin from Plutella xylostella, (designated as PxDef) was first cloned and characterized. Amino acid sequence analysis showed that the mature peptide owned characteristic six-cysteine motifs with predicted isoelectric point of 5.57, indicating an anionic defensin. Quantitative real-time polymerase chain reaction analysis showed that PxDef was significantly induced in epidermis, fat body, midgut and hemocytes after injection of heat-inactivated Bacillus thuringiensis, while such an induction was delayed by the injection of live B. thuringiensis in the 4th instar larvae of P. xylostella. Knocking down the expression of nuclear transcription factor Dorsal in P. xylostella by RNA interference significantly decreased the mRNA level of PxDef, and increased the sensitivity of P. xylostella larvae to the infection by live B. thuringiensis. The purified recombinant mature peptide (PxDef) showed higher activity against Gram-positive bacteria, with the minimum inhibition concentrations of 1.6 and 2.6 µM against B. thuringiensis and Bacillus subtilis, respectively. To our knowledge, this is the first report about an anionic PxDef, which may play an important role in the immune system of P. xylostella against B. thuringiensis.

MicroRNAs (miRNAs) are now recognized as key post-transcriptional regulators in regulation of phenotypic diversity. Qinlingacris elaeodes is a species of the alpine grasshopper, which is endemic to China. Adult individuals have three wing forms: wingless, unilateral-winged and short-winged. This is an ideal species to investigate the phenotypic plasticity, development and evolution of insect wings because of its case of unilateral wing form in both the sexes. We sequenced a small RNA library prepared from mesothoraxes of the adult grasshoppers using the Illumina deep sequencing technology. Approximately 12,792,458 raw reads were generated, of which the 854,580 high-quality reads were used only for miRNA identification. In this study, we identified 49 conserved miRNAs belonging to 41 families and 69 species-specific miRNAs. Moreover, seven miRNA*s were detected both for conserved miRNAs and species-specific miRNAs, which were supported by hairpin forming precursors based on polymerase chain reaction. This is the first description of miRNAs in alpine grasshoppers. The results provide a useful resource for further studies on molecular regulation and evolution of miRNAs in grasshoppers. These findings not only enrich the miRNAs for insects but also lay the groundwork for the study of post-transcriptional regulation of wing forms.

There is an ongoing relationship between host plants and herbivores. The nutrient substances and secondary compounds found in the host plant can not only impact the growth and development process of herbivores, but, more importantly, may also affect their survival and reproductive fitness. Vitellogenesis is the core process of reproductive regulation and is generally considered as a reliable indicator for evaluating the degree of ovarian development in females. Vitellogenin (Vg) plays a critical role in the synthesis and secretion of yolk protein. In this study, the full-length cDNA of the Vg gene in an alien invasive species, the nipa palm hispid beetle Octodonta nipae Maulik (Coleoptera: Chrysomelidae) (OnVg) was cloned and, the effect of host plant on the OnVg expression level and ovarian development was investigated. The results revealed that the OnVg was highly and exclusively expressed in adult females, but barely detectable in larvae, pupae and adult males. The relative expression level of OnVg and egg hatchability were much higher in females fed on Phoenix canariensis (their preferred host) than those fed on Phoenix roebelenii. A positive correlation relationship between OnVg expression and egg hatchability was also detected. Additionally, the anatomy of the female reproductive system showed that the ovaries of individuals fed on P. canariensis were considerably more developed than in females fed on P. roebelenii. The results may be applicable to many pest management situations through reproductive disturbance by alternating host plant species or varieties or by reproductive regulation through vitellogenesis mediated by specific endocrine hormones.

The study aimed to quantify the influence of common plant polyamines and tyramine on probing behaviour in the bird cherry-oat aphid (Rhopalosiphum padi L.). Electrical penetration graphs (DC) were used to monitor the probing and feeding behaviour of R. padi exposed to the amines agmatine, cadaverine, putrescine, spermidine, spermine and tyramine. The study results showed that the analyzed amines tended to shorten the stylet activity of aphids in the gels (as indicated by the g-C pattern), prolong the duration of non-probing behaviour (g-np pattern) and decrease salivation into the gels (g-E1pattern) and ingestion from the gels (g-G pattern). The 10 mM concentration of the studied amines, especially cadaverine, reduced or completely inhibited aphid ingestion. The obtained results demonstrate that plant amines participate in plant defence responses to R. padi through disturbance of its probing behaviour and the intensity of such effects is concentration dependent.

Necremnus tutae is native to the Mediterranean region where it has been observed in greenhouses parasitizing the invasive Tuta absoluta on tomato. The objective of the present study was to determine whether augmentative releases of N. tutae can improve existing biological control of T. absoluta based on predation by Nesidicoris tenuis. Two experiments were carried out, of which the first evaluated different N. tutae release rates (1 and 2 N. tutae m−2 week−1). The parasitoid reduced plant and fruit damage, especially at the higher rate. However, such reduction was considered insufficient given the large numbers of parasitoids needed and still unacceptable level of fruit damage. The second experiment focused on combining the most efficient rate of N. tutae of those evaluated during the first experiment, with the pre- and post-planting release of N. tenuis and supplemental additions of Ephestia kuehniella eggs. Addition of N. tutae decreased leaf damage by T. absoluta regardless the release method for N. tenuis, but the pre-plant release of N. tenuis alone was sufficient to prevent fruit damage by T. absoluta. This suggested that the addition of N. tutae may not be necessary to obtain satisfactory control of T. absoluta following pre-plant application of N. tenuis, although different options for using N. tutae in commercial crops may still be possible.

A generalist predator's ability to contribute to biological control is influenced by the decisions it makes during foraging. Predators often use flexible foraging tactics, which allows them to pursue specific types of prey at the cost of reducing the likelihood of capturing other types of prey. When a pest insect has low nutritional quality or palatability for a predator, the predator is likely to reject that prey in favour of pursuing alternative, non-pest prey. This is often thought to limit the effectiveness of generalist predators in consuming aphids, which are of low nutritional quality for many generalist predators. Here, we report behavioural assays that test the hypothesis that the generalist predator, Grammonota inornata (Araneae: Linyphiidae), preferentially forages for a non-pest prey with high nutritional quality (springtails), and rejects a pest prey with low nutritional quality (aphids). In no-choice assays, molecular gut-content analysis revealed that spiders continued to feed on the low-quality aphids at high rates, even when high-quality springtails were readily available. When provided a choice between aphids and springtails in two-way choice tests, spiders did not show the expected preference for springtails. Decision-making by spiders during foraging therefore appears to be sub-optimal, possibly because of attraction to the less frequently encountered of two preys as part of a dietary diversification strategy. These results indicate that behavioural preferences alone do not necessarily compromise the pest-suppression capacity of natural enemies: even nutritionally sub-optimal pest prey can potentially be subject to predation and suppression by natural enemies.