Insects, Volume 12, Issue 8 (August 2021) – 100 articles

Each fall, monarch butterflies in eastern North America undergo an extraordinary long-distance migration to wintering areas in central Mexico, where they remain until returning northward in the spring. Migrants survive the overwintering period by metabolizing lipid reserves accumulated exclusively though floral nectar; however, there is little known about how individuals maximize foraging efficiency in the face of floral environments that constantly change in complex and unpredictable ways along their migratory route. Here, a proboscis extension paradigm is used to investigate the role of cognition during the foraging phase of monarch migration. Male and female migratory butterflies were consecutively trained to discriminate between two color and odor cues and then tested for their ability to simultaneously retain the information on the reward value of each cue in memory without reinforcement over a period of 7 days. To gain further insight into cognitive abilities of monarchs as a migratory species, a second set of captive-reared males and females were tested under harnessed conditions at the same time as wild-caught fall migrants. Results showed that male and female migrants can learn the reward properties of color and odor cues with over 75% accuracy after less than 40 s of exposure and can simultaneously retain visual and olfactory information predicting the availability of floral rewards in memory without reinforcement for at least 7 days. Captive-reared male butterflies also showed the ability to retain visual and olfactory information in long-term memory for 7 days; however, 80% of captive-reared females could not retain color cues in long-term memory for more than 24 h. These novel findings are consistent with the view that monarch butterflies, as a migratory species, have enhancements to long-term memory that enable them to minimize the amount of time and energy wasted searching for suitable nectar sources during their annual fall migration, thereby optimizing migratory performance and increasing the chance of overwinter survival. The possibility that female monarchs undergo a seasonal change in visual long-term memory warrants further empirical investigation. Full article

We examined the contents of nutritional importance, i.e., amino acids, fatty acids and minerals of different developmental stages of drones of two honey bee subspecies, namely Apis mellifera carnica and A. m. mellifera. The results revealed that, in general, individual amino acid amounts and therefore the total protein increased along with the developmental stages of the drones. No statistically significant differences were found between the same developmental stages of the two subspecies. The reverse, i.e., a decrease with developmental stage occurred in relation to the fatty acid composition. Most of the minerals were higher at advanced developmental stages. Overall, the high protein content (31.4–43.4%), small amount of fat (9.5–11.5%) and abundance of minerals such asiron and zinc, make drones a suitable nutritional resource. Even though nutrient content, especially protein, was higher in the pupae than the prepupae, we propose prepupae also as a commercial product based on their higher biomass production. Provided standard production protocols maintaining hygiene and safety will be adhered to, we propose that drone honey bees can be utilized as human food or animal feed. Full article

Fall armyworm (FAW), Spodoptera frugiperda, is a major pest of maize in the Americas and recently invaded the Eastern hemisphere. It was first detected in India in 2018 and is considered a major threat to maize production. FAW control largely relies on the application of chemical insecticides and transgenic crops expressing Bacillus thuringiensis insecticidal proteins. Assessing FAW resistance and insecticide susceptibility is a cornerstone to develop sustainable resistance management strategies. In this study, we conducted more than 400 bioassays to assess the efficacy of nine insecticides from seven mode-of-action classes against 47 FAW populations collected in 2019 and 2020 across various geographical areas in India. The resistance status of the field-collected populations was compared to an Indian population sampled in 2018, and an insecticide susceptible reference population collected in 2005 in Brazil. Low to moderate resistance levels were observed for thiodicarb, chlorpyriphos, deltamethrin, chlorantraniliprole and flubendiamide in several populations (including the reference population collected in 2018). The highest resistance ratios were observed for deltamethrin which likely compromises recommended label rates for pyrethroid insecticides in general. Our data provide a useful baseline for future FAW resistance monitoring initiatives and highlight the need to implement insecticide resistance management strategies. Full article

To determine the significance of mitochondrial genome characteristics in revealing phylogenetic relationships and to shed light on the molecular evolution of the Coeliadinae species, the complete mitochondrial genomes (mitogenomes) of five Coeliadinae species were newly sequenced and analyzed, including Hasora schoenherr, Burara miracula, B. oedipodea, B. harisa, and Badamia exclamationis. The results show that all five mitogenomes are double-strand circular DNA molecules, with lengths of 15,340 bp, 15,295 bp, 15,304 bp, 15,295 bp, and 15,289 bp, respectively, and contain the typical 37 genes and a control region. Most protein-coding genes (PCGs) begin with ATN, with 3 types of stop codons including TAA, TAG, and an incomplete codon T-; most of the genes terminate with TAA. All of the transfer RNA genes (tRNAs) present the typical cloverleaf secondary structure except for the trnS1. Several conserved structural elements are found in the AT-rich region. Phylogenetic analyses based on three datasets (PCGs, PRT, and 12PRT) and using maximum likelihood (ML) and Bayesian inference (BI) methods show strong support for the monophyly of Coeliadinae, and the relationships of the five species are (B. exclamationis + ((B. harisa + (B. oedipodea + B. miracula)) + H. schoenherr)). Full article

Flavonoids detected in soybean Glycine max (L.) Merr. (Fabaceae) cause various alterations in the metabolism, behavior, and development of insect herbivores. The pea aphid Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae) poses potential threat to soybeans, but the effect of individual flavonoids on its feeding-associated behavior is relatively unknown. We monitored probing behavior (stylet penetration activities) of A. pisum on its preferred host plant, Pisum sativum L. untreated (control) and treated with 0.1% ethanolic solutions of flavonoids apigenin, daidzein, genistein, and kaempferol. We applied the electrical penetration graph (electropenetrography, EPG) technique, which visualizes the movements of aphid stylets within plant tissues. None of the applied flavonoids affected the propensity to probe the plants by A. pisum. However, apigenin enhanced the duration of probes in non-phloem tissues, which caused an increase in the frequency and duration of stylet mechanics derailment and xylem sap ingestion but limited the ingestion of phloem sap. Daidzein caused a delay in reaching phloem vessels and limited sap ingestion. Kaempferol caused a reduction in the frequency and duration of the phloem phase. Genistein did not affect aphid probing behavior. Our findings provide information for selective breeding programs of resistant plant cultivars to A. pisum. Full article

Cheatgrass is an annual grass species from Eurasia that has become invasive in much of western North America. It has been implicated in recent increases in the frequency, size, and intensity of wildfires, contributing to severe economic, environmental, and social destruction. In order to reduce this damage, the USDA-ARS established a classical biological control program against cheatgrass. In 2018 and 2019, adult gall midges were collected emerging from cheatgrass seed heads collected at several sites in Bulgaria and Greece; this is the first gall midge ever recorded from cheatgrass. Morphological comparisons with related midge species recorded from other plant hosts revealed that this midge from cheatgrass is a new species, described here as Stenodiplosis tectori n. sp. This status was supported by sequence comparisons of a barcode region of the gene encoding the mitochondrial cytochrome c subunit I (CO1) protein in Stenodiplosis tectori n. sp. and three congeners. The present study is the first to report MT-CO1 data in the genus Stenodiplosis. The ingroup Stenodiplosis tectori n. sp. collected in the Balkans grouped in one phylogenetic supported clade, with an average K2P-distance from its closest related congener, S. sorghicola, of 7.73% (SD = 1.10). The findings indicated relatively high year-to-year within-population diversity. Implications for this gall midge’s utility as a biological control agent of cheatgrass are discussed. Full article

In this study, the complete mitochondrial genomes (mitogenomes) of Hestina persimilis and Hestinalis nama (Nymphalidae: Apaturinae) were acquired. The mitogenomes of H. persimilis and H. nama are 15,252 bp and 15,208 bp in length, respectively. These two mitogenomes have the typical composition, including 37 genes and a control region. The start codons of the protein-coding genes (PCGs) in the two mitogenomes are the typical codon pattern ATN, except CGA in the cox1 gene. Twenty-one tRNA genes show a typical clover leaf structure, however, trnS1(AGN) lacks the dihydrouridine (DHU) stem. The secondary structures of rrnL and rrnS of two species were predicted, and there are several new stem loops near the 5′ of rrnL secondary structure. Based on comparative genomic analysis, four similar conservative structures can be found in the control regions of these two mitogenomes. The phylogenetic analyses were performed on mitogenomes of Nymphalidae. The phylogenetic trees show that the relationships among Nymphalidae are generally identical to previous studies, as follows: Libytheinae\Danainae + ((Calinaginae + Satyrinae) + Danainae\Libytheinae + ((Heliconiinae + Limenitidinae) + (Nymphalinae + (Apaturinae + Biblidinae)))). Hestinalisnama is apart from Hestina, and closely related to Apatura, forming monophyly. Full article

Studies were carried out in the laboratory to understand the optimum environmental conditions at which the ectoparasitoid, Habrobracon hebetor Say (Hymenoptera: Braconidae), can paralyze and lay eggs when reared on the larvae of the stored product pest, Plodia interpunctella Hübner (Lepidoptera: Pyralidae). At the four temperatures investigated (20, 25, 30, and 35 °C), optimum temperatures for oviposition were found to be 25 and 30 °C, while 35 °C was the least favorable temperature. No significant differences were found between the percentages of diapausing and non-diapausing larvae paralyzed by the wasp at the temperatures of 20, 25, 30, 35 °C within 5 days. However, in another experiment that investigated the effect of photoperiods at different temperatures that included 15, 19 and 28 °C, the number of paralyzed larvae was highly reduced at low temperatures (15 °C) but photoperiods had no significant impact on the number of host larvae paralyzed. In addition, observations at short time intervals also showed that lower temperatures slowed down host larvae paralysis. The results suggest that H. hebetor can paralyze host larvae of P. interpunctella more efficiently and deposit more eggs at temperatures within the range of 20–30 °C. Full article

Aedes aegypti is a significant vector for many tropical and subtropical flavivirus diseases. Only the female mosquito transmits pathogens, while the male plays a vital role in mating and species continuity. This study explored the total proteomes of females and males based on the physiological and genetic differences of female and male mosquitoes. Protein extracts from mosquitoes were analysed using LC–ESI–MS/MS for protein identification, protein interaction network analysis, functional ontology enrichment, and differential protein abundance analyses. Protein identification revealed 422 and 682 proteins exclusive to males and females, respectively, with 608 common proteins found in both sexes. The most significant PPIs (<1.0 × 10⁻¹⁶) were for common proteins, followed by proteins exclusive to females (<1.0 × 10⁻¹⁶) and males (1.58 × 10⁻¹²). Significant functional enrichments were observed in the biological process, molecular function, and cellular component for the male and female proteins. The abundance of the proteins differed, with one protein showing an increase (elongation factor 1 α, EF1α) and two showing reductions (actin family) in females versus males. Overall, the study verified the total proteomes differences between male and female Ae. aegypti based on protein identification and interactions, functional ontologies, and differentially abundant proteins. Some of the identified proteins merit further investigation to elucidate their roles in blocking viral transmission. Full article

Cuticular hydrocarbons (CHCs) cover insects’ bodies and play important roles in chemical communication, including nestmate recognition, for social insects. To enter colonies of a social host species, parasites may acquire host-specific CHCs or covertly maintain their own CHC profile by lowering its quantity. However, the chemical profile of small hive beetles (SHBs), Aethina tumida, which are parasites of honey bee, Apis mellifera, colonies, and other bee nests, is currently unknown. Here, adults of SHB and honey bee host workers were collected from the same field colonies and their CHC profiles were analysed using GC-MS. The chemical profiles of field-sampled SHBs were also compared with those of host-naive beetles reared in the laboratory. Laboratory-reared SHBs differed in their CHC profiles from field-sampled ones, which showed a more similar, but ten-fold lower, generic host CHC profile compared to host workers. While the data confirm colony-specific CHCs of honey bee workers, the profile of field-collected SHBs was not colony-specific. Adult SHBs often commute between different host colonies, thereby possibly preventing the acquisition of a colony-specific CHC profiles. An ester was exclusive to both groups of SHBs and might constitute an intraspecific recognition cue. Our data suggest that SHBs do not use any finely tuned chemical strategy to conceal their presence inside host colonies and instead probably rely on their hard exoskeleton and defence behaviours. Full article

In Quebec (Canada), viticulture has experienced steady growth in the last 35 years in terms of surfaces cultivated and value, although it is practiced in climatic conditions at the edge of what is considered a cool-climate area. This case study documents biodiversity studies conducted at the l’Orpailleur vineyard (Dunham, QC, Canada) from 1997 to 2021. In a first phase starting in 1997, the biodiversity of insecticide-free and insecticide-treated plots was determined for the taxa Scarabaeidae, Curculionidae, Chrysomelidae, Cicadellidae, Acari and Aranae. This step provided a baseline allowing to identify key arthropods. In a second phase starting in 2004, entomological issues were addressed on an ad hoc basis. In 2014, a third phase began with a perspective of sustainability and management of plant diversity in the vineyard to conserve natural enemies. Because of increased Japanese beetle (Popillia japonica-Scarabaeidae) populations and threats to vineyards, a biocontrol program based on the parasitoid Istocheta aldrichi (Tachinidae) was initiated. The unusually fast development of grapevines during the growing season, selection of flowering species, as well as selected arthropods associated with these flowering species, will be illustrated. Periodic update of protection programs will be required to address future challenges associated with climate change scenarios and world trade. Full article

The high fecundity of the most destructive pest Helicoverpa armigera and its great resistance risk to insecticides and Bt crops make the reproductive-destruction-based control of this pest extremely appealing. To find suitable targets for disruption of its reproduction, we observed the testis and ovary development of H. armigera and conducted deep sequencing of the ovary and testis small RNAs of H. armigera and quantitative RT-PCR (RT-qPCR) validation to identify reproduction-related micro RNAs (miRNAs). A total of 7,592,150 and 8,815,237 clean reads were obtained from the testis and ovary tissue, respectively. After further analysis, we obtained 173 novel and 74 known miRNAs from the two libraries. Among the 74 known miRNAs, 60 miRNAs existed in the ovary and 72 existed in the testis. Further RT-qPCR validation of 5 miRNAs from the ovary and 6 miRNAs from the testis confirmed 8 of them were indeed ovary- (miR-989a, miR-263-5p, miR-34) or testis-biased (miR-2763, miR-998, miR-2c, miR-2765, miR-252a-5p). The 8 ovary- or testis-biased miRNAs had a total of 30,172 putative non-redundant target transcripts, as predicted by miRanda and RNAhybrid. Many of these target transcripts are assigned to reproduction-related GO terms (e.g., oocyte maturation, vitellogenesis, spermatogenesis) and are members of multiple reproduction-related KEGG pathways, such as the JAK-STAT signaling pathway, oocyte meiosis, the insulin signaling pathway, and insect hormone biosynthesis. These results suggest that the 8 gonad-biased miRNAs play important roles in reproduction and may be used as the targets for the development of reproductive-destruction-based control of H. armigera and, possibly, other lepidopteran pests. Full article

Genome sequencing of a diverse array of arthropod genomes is already underway, and these genomes will be used to study human health, agriculture, biodiversity, and ecology. These new genomes are intended to serve as community resources and provide the foundational information required to apply ‘omics technologies to a more diverse set of species. However, biologists require genome annotation to use these genomes and derive a better understanding of complex biological systems. Genome annotation incorporates two related, but distinct, processes: Demarcating genes and other elements present in genome sequences (structural annotation); and associating a function with genetic elements (functional annotation). While there are well-established and freely available workflows for structural annotation of gene identification in newly assembled genomes, workflows for providing the functional annotation required to support functional genomics studies are less well understood. Genome-scale functional annotation is required for functional modeling (enrichment, networks, etc.). A first-pass genome-wide functional annotation effort can rapidly identify under-represented gene sets for focused community annotation efforts. We present an open-source, open access, and containerized pipeline for genome-scale functional annotation of insect proteomes and apply it to various arthropod species. We show that the performance of the predictions is consistent across a set of arthropod genomes with varying assembly and annotation quality. Full article

Tick cell culture facilitates research on the biology of ticks and their role as vectors of pathogens that affect humans, domestic animals, and wildlife. Because two-dimensional cell culture doesn’t promote the development of multicellular tissue-like composites, we hypothesized that culturing tick cells in a three-dimensional (3-D) configuration would form spheroids or tissue-like organoids. In this study, the cell line BmVIII-SCC obtained from the cattle fever tick, Rhipicephalus (Boophilus) microplus (Canestrini, 1888), was cultured in different synthetic scaffold systems. Growth of the tick cells on macrogelatinous beads in rotating continuous culture system bioreactors enabled cellular attachment, organization, and development into spheroid-like aggregates, with evidence of tight cellular junctions between adjacent cells and secretion of an extracellular matrix. At least three cell morphologies were identified within the aggregates: fibroblast-like cells, small endothelial-like cells, and larger cells exhibiting multiple cytoplasmic endosomes and granular vesicles. These observations suggest that BmVIII-SCC cells adapted to 3-D culture retain pluripotency. Additional studies involving genomic analyses are needed to determine if BmVIII-SCC cells in 3-D culture mimic tick organs. Applications of 3-D culture to cattle fever tick research are discussed. Full article

Amino acids play a vital role in several biological processes in organisms and are mainly acquired through diet by most insects. The amino acid auxin permease (AAAP) transporter family is an important amino acid transporter gene family in insects for the transportation of amino acids into and out of cells across the plasma membrane. Here, we identified 21 putative AAAP family members in the genome of the brown planthopper (BPH), Nilaparvata lugens, a devastating pest that feeds only on the phloem sap of rice plants. Molecular characteristic analysis indicated large variations in protein features and amino acid sequences among the predicted AAAP family members in BPH. Phylogenetic analysis clustered these AAAP transporters into three subgroups, with the members in the same group sharing a similar pattern of conserved motif distribution. Through ortholog gene recognition and spatiotemporal gene expression analysis, the AAAP gene NlAAAP07, which was predicted to regulate BPH larval growth and female fecundity, was identified. RNA interference (RNAi)-mediated suppression of NlAAAP07 significantly postponed the duration of 3rd instar nymphs developing into adults from 7.4 days to 9.0 days, and decreased the oviposition amount and egg hatching rate of females by 30.7% and 11.0%, respectively. Our results provide a foundation for further functional analysis of AAAP transporters in BPH. Full article

In insects, neuropeptides and their receptors not only play a critical role in insect physiology and behavior but also are the potential targets for novel pesticide discoveries. Aphidius gifuensis is one of the most important and widespread aphid parasitoids, and has been successfully used to control aphid. In the present work, we systematically identified neuropeptides and their receptors from the genome and head transcriptome of A. gifuensis. A total of 35 neuropeptide precursors and 49 corresponding receptors were identified. The phylogenetic analyses demonstrated that 35 of these receptors belong to family-A, four belong to family-B, two belong to leucine-rich repeat-containing GPCRs, four belong to receptor guanylyl cyclases, and four belong to receptor tyrosine kinases. Oral ingestion of imidacloprid significantly up-regulated five neuropeptide precursors and four receptors whereas three neuropeptide precursors and eight receptors were significantly down-regulated, which indicated that these neuropeptides and their receptors are potential targets of some commercial insecticides. The RT-qPCR results showed that dopamine receptor 1, dopamine receptor 2, octopamine receptor, allatostatin-A receptor, neuropeptides capa receptor, SIFamide receptor, FMRFamide receptor, tyramine receptor and short neuropeptide F predominantly were expressed in the head whilst the expression of ion transport peptide showed widespread distribution in various tissues. The high expression levels of these genes suggest their important roles in the central nervous system. Taken together, our study provides fundamental information that may further our understanding of neuropeptidergic signaling systems in the regulation of the physiology and behavior of solitary wasps. Furthermore, this information could also aid in the design and discovery of specific and environment-friendly insecticides. Full article

The reproduction of aphids depends to a great extent on their host plants, an integration that impacts on the successful expansion of overwintering populations. Therefore, a survey was conducted to evaluate the globally distributed Capsella bursa-pastoris as an overwintering host of economically important aphid species, their parasitoids and hyperparasitoids in the southern and western regions of Turkey from November to March in 2006 to 2013. During this survey, 395 samples of C. bursa-pastoris were collected with 25 aphid species recorded. Among aphids that feed on this host, Myzus persicae, Aphis gossypii, Rhopalosiphum padi, Aphis fabae, Aphis craccivora, Lipaphis erysimi, and Brevicoryne brassicae were the most frequently recorded. In total, 10,761 individual parasitoids were identified. Binodoxys angelicae, Aphidius colemani, Aphidius matricariae, Diaeretiella rapae, Ephedrus persicae, and Lysiphlebus confusus were the most abundant aphidiines that emerged from the aphids collected from C. bursa-pastoris. Alloxysta spp. (Hymenoptera: Cynipoidea), Chalcidoidea (unidentified at genus level), and Dendrocerus spp. (Hymenoptera: Ceraphronoidea) were identified as hyperparasitoids on the parasitoids. These findings indicate that C. bursa-pastoris is a key non-agricultural plant that significantly contributes to the overwintering of numerous aphids and their parasitoids, which should be given serious consideration when biological control strategies are designed. Full article

β-1,3-glucan recognition proteins (βGRPs) as pattern recognition receptors (PRRs) play an important role in recognizing various pathogens and trigger complicated signaling pathways in insects. In this study, we identified a Bombyx mori β-1,3-glucan recognition protein gene named BmβGRP4, which showed differential expression, from a previous transcriptome database. The full-length cDNA sequence was 1244 bp, containing an open reading frame (ORF) of 1128 bp encoding 375 amino acids. BmβGRP4 was strongly expressed in the larval stages and highly expressed in the midgut of B. mori larvae in particular. After BmNPV infection, the expression of BmβGRP4 was reduced significantly in the midgut. Furthermore, a significant increase in the copy number of BmNPV was observed after the knockdown of BmβGRP4 in 5th instar larvae, while the overexpression of BmβGRP4 suppressed the proliferation of BmNPV in BmN cells. Subsequently, the expression analysis of several apoptosis-related genes and observation of the apoptosis morphology demonstrated that overexpression of BmβGRP4 facilitated apoptosis induced by BmNPV in BmN cells. Moreover, BmβGRP4 positively regulated the phosphatase and tensin homolog gene (BmPTEN), while expression of the inhibitor of apoptosis gene (BmIAP) was negatively regulated by BmβGRP4. Hence, we hypothesize that BmNPV infection might suppress BmPTEN and facilitate BmIAP to inhibit cell apoptosis by downregulating the expression of BmβGRP4 to escape host antiviral defense. Taken together, these results show that BmβGRP4 may play a role in B. mori response to BmNPV infection and lay a foundation for studying its functions. Full article

The silverleaf whitefly, Bemisia tabaci (Gennadius, Hemiptera: Aleyrodidae), is a major threat to field and horticultural crops worldwide. Persistent use of insecticides for the management of this pest is a lingering problem. In the present study, the status of sensitivity of B. tabaci to two neonicotinoids, imidacloprid and thiamethoxam, was evaluated. The expression pattern of two cytochrome P450 (cyp) genes and changes in the relative amount of symbionts in insecticide-treated B. tabaci were also assessed. Quantitative PCR (qPCR) studies indicate that the CYP6CM1 and CYP6CX1 genes were always expressed higher in imidacloprid-treated whitefly, suggesting a correlation between gene expression and the insect’s ability to detoxify toxic compounds such as insecticides. In addition, the thiamethoxam-treated population harbored higher Portiera and lower Rickettsia titers, whereas the imidacloprid-treated population harbored more Rickettsia at different time intervals. Interestingly, we also examined that an increase in exposure to both the insecticides resulted in a reduction in the mutualistic partners from their insect host. These differential responses of endosymbionts to insecticide exposure imply the complex interactions among the symbionts inside the host insect. The results also provide a deeper understanding of the molecular mechanism of resistance development that might be useful for formulating effective management strategies to control B. tabaci by manipulating symbionts and detoxifying genes. Full article

Aphid odorant-binding protein 9 is almost exclusively expressed in antennae and is well conserved between different aphid species. In order to investigate its function, we have expressed this protein and measured ligand-binding affinities to a number of common natural compounds. The best ligands are long-chain aldehydes and alcohols, in particular Z9-hexadecenal and Z11-hexadecenal, as well as 1-hexadecanol and Z11-1-hexadecenol. A model of this protein indicated Lys37 as the residue that is likely to establish strong interactions with the ligands, probably a Schiff base with aldehydes and a hydrogen bond with alcohols. Indeed, when we replaced this lysine with a leucine, the mutated protein lost its affinity to both long aldehydes and alcohols, while the binding of other volatiles was unaffected. Long-chain linear alcohols are common products of molds and have been reported as aphid antifeedants. Corresponding aldehydes, instead, are major components of sex pheromones for several species of Lepidoptera. We speculate that aphids might use OBP9 to avoid mold-contaminated plants as well as competition with lepidopteran larvae. Full article

(1) Modern, intensive agricultural practices have been attributed to the loss of insect biodiversity and abundance in agroecosystems for the last 80 years. The aim of this work is to test whether there are statistically significant differences in insect abundance between different zones and over time on the vineyard field. (2) The study was carried out in five intensive wine farms in Spain over a three-year period (2013–2015). Each field was divided into two zones, one where cover plants were planted, and another remained unchanged (without cover). (3) A clear trend to increase the average number of insect species and individuals throughout the years in all farms was observed. Moreover, the zones with cover plants showed a significant difference with respect to the zones without. (4) The use of permanent cover plants allows creating areas of refuge for the insects favouring their conservation and reducing the agriculture impact in the insect decline. Full article

Diapause is one of the survival strategies of insects for confronting adverse environmental conditions. Bombyx mori displays typical embryonic diapause, and offspring diapause depends on the incubation environment of the maternal embryo in the bivoltine strains of the silkworm. However, the molecular mechanisms of the diapause induction process are still poorly understood. In this study, we compared the differentially expressed miRNAs (DEmiRs) in bivoltine silkworm embryos incubated at diapause- (25 °C) and non-diapause (15 °C)-inducing temperatures during the blastokinesis (BK) and head pigmentation (HP) phases using transcriptome sequencing. There were 411 known miRNAs and 71 novel miRNAs identified during the two phases. Among those miRNAs, there were 108 and 74 DEmiRs in the BK and HP groups, respectively. By the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the predicted target genes of the DEmiRs, we found that aside from metabolism, the targets were also enriched in phototransduction-fly and insect hormone biosynthesis in the BK group and the HP group, respectively. Dual luciferase reporter assay illustrated that bmo-miR-6497-3p directly regulated Bmcycle and subsequently regulated the expression of circadian genes. These results imply that microRNAs, as vitally important regulators, respond to different temperatures and participate in the diapause induction process across species. Full article

Innate immune responses are essential to maintaining insect and tick health and are the primary defense against pathogenic viruses, bacteria, and fungi. Cell line research is a powerful method for understanding how invertebrates mount defenses against pathogenic organisms and testing hypotheses on how these responses occur. In particular, immortal arthropod cell lines are valuable tools, providing a tractable, high-throughput, cost-effective, and consistent platform to investigate the mechanisms underpinning insect and tick immune responses. The research results inform the controls of medically and agriculturally important insects and ticks. This review presents several examples of how cell lines have facilitated research into multiple aspects of the invertebrate immune response to pathogens and other foreign agents, as well as comments on possible future research directions in these robust systems. Full article

Background: The destructive insect pest Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae) is a polyphagous species targeting many economically important plants. The extensive and arbitrary use of insecticides has resulted in the build-up of insecticide resistance and pesticide residues accumulating in food. Therefore, it is becoming evident that alternative pest management tools are needed to reduce risks to humans, the environment, and non-target organisms, and at the same time, they should be used in field application at the lowest cost. Methods: In view of this objective, the present study demonstrates the toxicity of lemongrass (Cymbopogon citratus (DC.) Stapf) essential oil (EO), against the black cutworm A. ipsilon under controlled laboratory conditions in terms of measuring the activity of peroxidase and detoxification enzymes. The chemical components of the EO were analyzed using GC–MS. Results: The results show that after 96 h post treatment, the LC₁₅ and LC₅₀ values were 427.67 and 2623.06 mg/L, respectively, of C. citratus EO on second-instar larvae of A. ipsilon. A slight significance in elongation of the larval duration with LC₁₅ and LC₅₀ value was found with control. By GC–MS analysis, the main compounds identified in the EO were α-citral and β-citral with percentages of 35.91%, and 35%, respectively. The oxidative stress indicates a significant increase in CAT and lipid peroxidase enzyme activity after 96 h post treatment at the LC₁₅ and LC₅₀. Conversely, the detoxification enzyme activity shows an inhibition of CarE and GST enzymes of larvae exposed to LC₁₅ and LC₅₀ values in response to C. citratus EO. Conclusions: The present data show that lemongrass EO has insecticidal activity against the black cutworm, A. ipsilon. Full article

Tetranychus urticae is an important agricultural pest that feeds on more than 1100 plant species. To investigate gene expression network in development process of deutonymph, a comprehensive transcriptome analysis of different developmental time points of deutonymph in T. urticae was performed. Comparing with expression profile of 7 h, 309, 876, 2736, and 3432 differential expression genes were detected at time points 14 h, 21 h, 28 h, and 35 h, respectively. The expression dynamic analysis indicated that genes in hormone- (ecdysteroid and juvenile hormone) and cuticle- (chitin and cuticle proteins) related pathways were indispensable for development process in deutonymph. Among hormone related pathway genes, the ecdysteroid biosynthesis pathway genes were highly expressed at the growth period of development process, which is opposite to the expression patterns of juvenile hormone biosynthesis pathway genes. For cuticle related pathway genes, 13 chitinase genes were identified in the genome of T. urticae, and 8 chitinase genes were highly expressed in different time points of developmental process in the deutonymph of T. urticae. Additionally, 59 cuticle protein genes were identified from genome, and most of the cuticle protein genes were expressed in the molting period of developmental process in deutonymph. This study reveals critical genes involved in the development process of deutonymph and also provides comprehensive development transcriptome information for finding more molecular targets to control this pest. Full article

Citrus tristeza virus (CTV) is one of the most important citrus tree viruses: a graft-transmissible virus that can be vectored by several aphid species. Diaphorina citri is the insect vector of “Candidatus Liberibacter spp.”, a bacterium associated with citrus Huanglongbing (HLB). However, no detailed description of the relationship between CTV and D. citri has been reported. In this study, D. citri adults collected from CTV-infected “Shatangju” mandarin, “Newhall” sweet orange, and “fingered citron” trees in different orchards yielded CTV-positive rates of 40%, 65%, and 95%, respectively, upon detection by conventional PCR. Illumina HiSeq sequencing followed by de novo assembly recovered the primary full CTV genome from the RNA of 30 D. citri adults sampled from CTV-positive citrus plants. Molting and adult emergence did not affect the presence or titers of CTV within the D. citri; however, the persistence of CTV in psyllids varied among different host plant species. Groups of 10 D. citri (from a population 85% CTV-positive) were shown to potentially transmit CTV to two citrus species, “Shatangju” mandarin and “Eureka” lemon, yielding 58.33% and 83.33% CTV-positive plants, respectively. No transmission of CTV to orange jasmine plants occurred. Thus, this study reports on the ability of D. citri to acquire and transmit CTV, making D. citri as a vector of two important citrus pathogens, warranting further attention and investigation. Full article

Nocturnal light pollution from anthropogenic origin is increasing worldwide and is recognised as a major threat for nocturnal biodiversity. We studied the impact of artificial light on the mate attraction success of female common glow-worms (Lampyris noctiluca L.) by daily monitoring their glowing status in the field, acting as a proxy for mating status throughout the mating season. We found that females in dark surroundings typically stopped glowing after one night, indicating that they had mated, while females in illuminated areas glowed for significantly more nights, in some cases up to 15 nights. Our study confirms previous findings and hypotheses that females exposed to artificial light suffer from a reduced mate attraction success with a negative impact on populations. Full article

The attraction range of YATLORf pheromone traps to adults of four species of Agriotes (A. brevis, A. sordidus, A. litigiosus, and A. ustulatus) was studied to provide additional information about the most harmful Agriotes species in Europe. Male click beetles were marked and released at different distances from a pheromone trap. The recapture rate was calculated and analyzed using analysis of variance. The recapture rate was significantly affected by distance, species, and wind direction. The recapture rate decreased as distance increased. The majority of beetles were caught from short distances (up to 10 m) within the first five days. A. brevis, a mainly crawling species, showed the lowest recapture rate. The wind direction affected the recovery rate, with a significantly lower number of beetles moving downwind from the release points. Maximum sampling ranges and effective sampling areas were calculated. The obtained estimations were low (53 to 86 m and 509 to 2602 m², respectively) for all the considered Agriotes species, suggesting that they were unsuitable for use as mass trapping instruments to disrupt mating. However, it seems possible to use the traps not only as monitoring tools, but also as attract-and-kill strategies for most beetle populations. Full article

Investigating the distributions of invasive species in marginal habitats can give clues to the factors constraining invasive spread. Vespula germanica is the most widely distributed of all the invasive Vespids, which in the Southern Hemisphere typically have large extensive invasive populations. In contrast, the invasion into South Africa has been slow and is still confined to a small geographic area. Here we analyse the distribution of all recent nest records in South Africa (n = 405). The distance to main rivers, mean annual rainfall, summer normalised difference moisture index (NDMI) values, and mean annual temperatures (average, minimum, maximum, and summer maximum temperature) was measured for every nest. We find that value ranges of these variables are different between the value ranges recorded for nests, the general distribution area of the wasp, and the area of absence. Optimised Hot Spot Analysis was used to quantify spatial structure in the measured climatic variables. Generally, factors related to moisture stress set the environmental limits of V. germanica’s landscape distribution. Due to the strong preference of nesting sites close to river courses, for higher rainfall conditions, medium to medium-high NDMI values, and lower mean annual temperatures, it is unlikely that V. germanica will be able to spread uniformly where it is currently found in South Africa. Full article

Varroa destructor is considered one of the most devastating parasites of the honey bee, Apis mellifera, and a major problem for the beekeeping industry. Currently, the main method to control Varroa mites is the application of drugs that contain different acaricides as active ingredients. The pyrethroid tau-fluvalinate is one of the acaricides most widely used in beekeeping due to its efficacy and low toxicity to bees. However, the intensive and repetitive application of this compound produces a selective pressure that, when maintained over time, contributes to the emergence of resistant mites in the honey bee colonies, compromising the acaricidal treatments efficacy. Here we studied the presence of tau-fluvalinate residues in hives and the evolution of genetic resistance to this acaricide in Varroa mites from honey bee colonies that received no pyrethroid treatment in the previous four years. Our data revealed the widespread and persistent tau-fluvalinate contamination of beeswax and beebread in hives, an overall increase of the pyrethroid resistance allele frequency and a generalized excess of resistant mites relative to Hardy–Weinberg equilibrium expectations. These results suggest that tau-fluvalinate contamination in the hives may seriously compromise the efficacy of pyrethroid-based mite control methods. Full article