Published by MDPI
Online ISSN: 2075-4450
The Asian tiger mosquito, Aedes albopictus, is a competent vector for arboviruses and recently was implicated as the vector of the first autochthonous cases of dengue and chikungunya in southern Europe. The objective of this study was to analyze the flight performance of female Ae. albopictus of different ages that were starved, sugar-fed, or sugar-fed and blood-fed, using flight mills. After three days of starvation post emergence, females flew an average distance of 0.7 ± 0.5 km in 1.9 ± 1.5 h during a 16 h trial period, whereas sugar- or sugar- and blood-fed females of this age covered a significantly higher distance of around 3 km with a mean total flight time of around 6 h. The age of females (up to four weeks) had no effect on performance. The average of maximal continuous flight segments of sugar-fed (2.14 ± 0.69 h) and blood-fed (3.17 ± 0.82 h) females was distinctly higher than of starved females (0.38 ± 0.15 h) of which most flyers (83%) performed maximal flight segments that lasted no longer than 0.5 h. Overall, the results for the laboratory monitored flight performance of Ae. albopictus confirm their ability to disperse a few kilometres between breeding site and host.
Paralytic effects of ingested Bt on various lepidopteran families and species. 
This review focuses on common insecticidal virulence factors from entomopathogenic bacteria with special emphasis on two insect pathogenic bacteria Photorhabdus (Proteobacteria: Enterobacteriaceae) and Bacillus (Firmicutes: Bacillaceae). Insect pathogenic bacteria of diverse taxonomic groups and phylogenetic origin have been shown to have striking similarities in the virulence factors they produce. It has been suggested that the detection of phage elements surrounding toxin genes, horizontal and lateral gene transfer events, and plasmid shuffling occurrences may be some of the reasons that virulence factor genes have so many analogs throughout the bacterial kingdom. Comparison of virulence factors of Photorhabdus, and Bacillus, two bacteria with dissimilar life styles opens the possibility of re-examining newly discovered toxins for novel tissue targets. For example, nematodes residing in the hemolymph may release bacteria with virulence factors targeting neurons or neuromuscular junctions. The first section of this review focuses on toxins and their context in agriculture. The second describes the mode of action of toxins from common entomopathogens and the third draws comparisons between Gram positive and Gram negative bacteria. The fourth section reviews the implications of the nervous system in biocontrol.
MON 810 maize was developed against Ostrinia nubilalis and is suggested to indirectly decrease Fusarium spp. infestation in maize ears. To evaluate this effect, co-occurrence of insect and fungal pests on MON 810 maize was studied. During 2009, exceptionally high maize ear infestation occurred in Julianna-major (Hungary). From investigation of some thousands of maize ears, the majority of the larval damage originated from Helicoverpa armigera larvae, while O. nubilalis larvae contributed significant damage only at a single plot. Fusarium verticillioides infection appeared only in a small portion (~20–30%) of the insect damaged cobs. H. armigera and O. nubilalis larvae feeding on F. verticillioides mycelia can distribute its conidia with their fecal pellets. MON 810 maize showed 100% efficacy against O. nubilalis in the stem, but lower efficacy against O. nubilalis and H. armigera in maize ears. The ~Cry1Ab toxin content of maize silk, the entry site of H. armigera, was lower than that in the leaves/stem/husk leaves of MON 810. Fusarium-infected MON 810 cobs are rarely found and only after larval damage by O. nubilalis. H. armigera larvae could not tolerate well F. verticillioides infected food and attempted to move out from the infected cobs. For further feeding they re-entered the maize ears through the 8–12 husk leaves, but in the case of the MON 810 variety, they usually could not reach the kernels. Apical damage on cobs resulted in only a minor (about one-tenth of the cob) decrease in yield.
Conventional thought would suggest that there are sufficient venues for the publication of entomological scientific inquiries and begs the question: “Why begin another outlet for insect-related scientific comment?” The open access journal Insects is a response to the pressing global thirst for information and an acknowledgement of the potential of electronic media. Dissemination of information is critical to growing the global knowledge base through easy and open access. This new journal is a celebration of the diversity of insects, other arthropods, and their relationship with humans in our shared environment. [...]
Map showing the different Antarctic zones and locations referred to in the text. Each zone is indicated by colour: yellow corresponds to sub-Antarctica, white to maritime Antarctica, and grey to continental Antarctica. In addition, the ‘Gressitt Line’ is shown at the base of the Antarctic Peninsula(see text). 
Summary of phylogeographic analyses performed to date on springtails in
Figure showing the phylogeographic sampling that has been achieved for four Antarctic springtail species across their distributional ranges (as indicated by vertical lines beneath/above species names). The codes used in the two inset boxes are location codes and refer to potential refugial locations for each species as referred to in the respective publications and Table 1. Dashed line in the inset boxes indicate heavy or fine biogeographic breaks among regions and in many cases also represent major glacial systems. See text for further details. 
ArcGIS Map of terrestrial invertebrate occurrence records and ice-free terrain [104,105]. Areas shaded in black represent the ice-free terrain where no terrestrial invertebrate collections have been made; red triangles illustrate all springtail records while green circles illustrate all other terrestrial invertebrate records. 
We review current phylogeographic knowledge from across the Antarctic terrestrial landscape with a focus on springtail taxa. We describe consistent patterns of high genetic diversity and structure among populations which have persisted in glacial refugia across Antarctica over both short (10 Mya) timescales. Despite a general concordance of results among species, we explain why location is important in determining population genetic patterns within bioregions. We complete our review by drawing attention to the main limitations in the field of Antarctic phylogeography, namely that the scope of geographic focus is often lacking within studies, and that large gaps remain in our phylogeographic knowledge for most terrestrial groups.
(a) Parapatric distribution of chromosomal races of the viatica species group in southeastern Australia proposed by White et al. [21,23]. An inset shows distribution of three races on Kangaroo Island. A 100 m isobath is indicated as a proxy of an ancient coastline at glacial maxima during the Pleistocene. Karyotypes of each race ( ♂/♀) are: viatica 19, 2n = 19/20, XO/XX; viatica 17, 2n = 17/18, XO/XX; P24(XO), 2n = 17/18, XO/XX; P24(XY), 2n = 16, XY/XX; P24(XY)-Translocation, 2n = 16, XY/XX; P25(XO), 2n = 19/20, XO/XX; P25(XY), 2n = 18, XY/XX; P45b(XO), 2n = 19/20, XO/XX; P45b(XY), 2n = 18, XY/XX; P50, 2n = 19/20, XO/XX; V. pichirichi , 2n = 19/20, XO/XX. (b) Thirteen genetic clusters resolved by the Bayesian clustering analysis using 35 allozyme loci, superimposed on a distribution map. Red circles with solid line indicate clusters shared among multiple chromosomal races. Four taxa [P24(XY), P24(XY)-Translocation, P45c, and V. pichirichi ] comprise exclusive genetic clusters (blue circles with dashed line). 
Estimated allele frequency clines of chromosomal (dotted red line), nuclear (solid black line) and mitochondrial markers (dashed blue line) across a hybrid zone between P24(XY) (to the left side) and viatica 17 (to the right side) on Kangaroo Island based on maximum likelihood models. Best fit models are the sigmoid model for the chromosomal and mitochondrial markers and asymmetrical stepped model for the nuclear markers. Circles, squares, and triangles represent observed allele frequencies of chromosomal, nuclear (average of 10 autosomal loci), and mitochondrial markers, respectively. Distance is expressed relative to the center (= 0 km) of the average nuclear cline. 
Chromosomal rearrangements can alter the rate and patterns of gene flow within or between species through a reduction in the fitness of chromosomal hybrids or by reducing recombination rates in rearranged areas of the genome. This concept, together with the observation that many species have structural variation in chromosomes, has led to the theory that the rearrangements may play a direct role in promoting speciation. Australian morabine grasshoppers (genus Vandiemenella, viatica species group) are an excellent model for studying the role of chromosomal rearrangement in speciation because they show extensive chromosomal variation, parapatric distribution patterns, and narrow hybrid zones at their boundaries. This species group stimulated development of one of the classic chromosomal speciation models, the stasipatric speciation model proposed by White in 1968. Our population genetic and phylogeographic analyses revealed extensive non-monophyly of chromosomal races along with historical and on-going gene introgression between them. These findings suggest that geographical isolation leading to the fixation of chromosomal variants in different geographic regions, followed by secondary contact, resulted in the present day parapatric distributions of chromosomal races. The significance of chromosomal rearrangements in the diversification of the viatica species group can be explored by comparing patterns of genetic differentiation between rearranged and co-linear parts of the genome.
Blood-feeding insects have evolved resistance to various insecticides (organochlorines, pyrethroids, carbamates, etc.) through gene mutations and increased metabolism. Bed bugs (Cimex lectularius) are hematophagous ectoparasites that are poised to become one of the major pests in households throughout the United States. Currently, C. lectularius has attained a high global impact status due to its sudden and rampant resurgence. Resistance to pesticides is one factor implicated in this phenomenon. Although much emphasis has been placed on target sensitivity, little to no knowledge is available on the role of key metabolic players (e.g., cytochrome P450s and glutathione S-transferases) towards pesticide resistance in C. lectularius. In this review, we discuss different modes of resistance (target sensitivity, penetration resistance, behavioral resistance, and metabolic resistance) with more emphasis on metabolic resistance.
The time-lapse photography system and other equipment used to track the behavioral movement of the bed bugs throughout the experimental arena. 
Sample walking-path of one bed bug, within the experimental arena (91.44 × 91.44 cm), demonstrating characteristic host-seeking behavior in the (a) presence and (b) absence of host breath. Tracks began (white cross) at the harborage in the center of the arena. In both instances, the breathing tube (black dot) was located in the upper left corner of the arena. The vertical and horizontal axes are measured in centimeters.
Orientation response of bed bugs in the four treatment groups: females in the presence of host breath (a), females in the absence of host breath (b), males in the presence of host breath (c), males in the absence of host breath (d). The rose diagram is a circular histogram, measured in degrees, progressing clockwise from the location of the breathing tube at 0° . Dark bars radiating from the center outward indicate the weighted mean vector, α W , of the bugs, with longer bar indicating a higher frequency of bugs and shorter bars a lower frequency of bugs. If Rayleigh test showed significant deviation from circular uniformity (P < 0.05), then the average α W ( white line ) of the treatment group is shown 
The reemergence of the bed bug, Cimex lectularius Linnaeus, has recently spawned a frenzy of public, media, and academic attention. In response to the growing rate of infestation, considerable work has been focused on identifying the various host cues utilized by the bed bug in search of a meal. Most of these behavioral studies examine movement within a confined environment, such as a Petri dish. This has prevented a more complete understanding of the insect’s host-seeking process. This work describes a novel method for studying host-seeking behavior, using various movement parameters, in a time-lapse photography system. With the use of human breath as an attractant, we qualitatively and quantitatively assessed how bed bugs navigate their environment between its harborage and the host. Levels of behavioral activity varied dramatically between bed bugs in the presence and absence of host odor. Bed bugs demonstrated not simply activation, but attraction to the chemical components of breath. Localized, stop-start host-seeking behavior or alternating periods of movement and pause were observed among bed bugs placed in the environment void of human breath, while those exposed to human breath demonstrated long range, stop-start host-seeking behavior. A more comprehensive understanding of bed bug host-seeking can lead to the development of traps and monitors that account for unique subtleties in their behavior. The time-lapse photography system uses a large, artificial environment and could also be employed to study other aspects of the insect’s behavioral patterns.
White pine weevil (Pissodes strobi, Peck.) is a native forest insect pest in the Pacific Northwest of North America that attacks species of spruce (Picea spp.) and pine (Pinus spp.). Young Sitka spruce [Picea sitchensis (Bong.) Carr.] trees are particularly susceptible to weevil attack. Pockets of naturally occurring Sitka spruce resistance have been identified in high weevil hazard areas in coastal British Columbia. In this study, we characterize behavioral, physiological and reproductive responses of weevils to an extremely resistant Sitka spruce genotype (H898) in comparison to a highly susceptible genotype (Q903). The experiments relied on a large number of three-year-old clonally propagated trees and were therefore restricted to two contrasting Sitka spruce genotypes. When exposed to resistant trees, both male and female weevils were deterred during host selection and mating, females showed delayed or reduced ovary development, and successful reproduction of weevils was prevented on resistant trees.
Use of the digital microscope for in situ measurement of maize pollen in the field. The microscope is powered via the USB hub, the labtop monitor serves to verify the images taken. 
Image of upper leaf side of Centaurea scabiosa taken in situ with Dino-Lite microscope (200×). Maize pollen can be distinguished from other pollen species by its relatively large size between 80–120 μm, color, shape and structure of the exine. Measurements are given by Dino Capture software. 
Images taken in situ with Dino-Lite microscope (200×), upper leaf side. ( a ) Zea mays 540 pollen/cm 2 ; ( b ) Chenopodium album , 2800 pollen/cm 2 ; ( c ) Rumex spp., 
Images taken in situ with Dino-Lite microscope. ( a , b ) Examples of pollen accumulation on Urtica dioica on upper leaf side (50×). ( c ) Leaf underside of Urtica dioica (50×); ( d ) Sideways view on leaf of Succisa scabiosa (200×). 
Maize is wind pollinated and produces huge amounts of pollen. In consequence, the Cry toxins expressed in the pollen of Bt maize will be dispersed by wind in the surrounding vegetation leading to exposure of non-target organisms (NTO). NTO like lepidopteran larvae may be affected by the uptake of Bt-pollen deposited on their host plants. Although some information is available to estimate pollen deposition on host plants, recorded data are based on indirect measurements such as shaking or washing off pollen, or removing pollen with adhesive tapes. These methods often lack precision and they do not include the necessary information such as the spatial and temporal variation of pollen deposition on the leaves. Here, we present a new method for recording in situ the amount and the distribution of Bt-maize pollen deposited on host plant leaves. The method is based on the use of a mobile digital microscope (Dino-Lite Pro, including DinoCapture software), which can be used in combination with a notebook in the field. The method was evaluated during experiments in 2008 to 2010. Maize pollen could be correctly identified and pollen deposition as well as the spatial heterogeneity of maize pollen deposition was recorded on maize and different lepidopteran host plants (Centaurea scabiosa, Chenopodium album, Rumex spp., Succina pratensis and Urtica dioica) growing adjacent to maize fields.
Bioinformatics analysis of BmβGRP4 cDNA sequence. (A) Complete nucleotide sequence and deduced amino acid sequence of BmβGRP4. Numbers on the left side represent nucleotide and amino acid positions. The initiation codon (ATG) and termination codon (TAA) are indicated in black italics. The signal peptide is represented in the black box and the Glyco_hydro_16 domain is highlighted by a single line. (B) Structural domain of BmβGRP4 predicted using SMART software. The red box indicates the signal peptide and the blue box represents the Glyco_hydro_16 domain. (C) Phylogenetic relationships of BmβGRP4 in different species using the neighbor-joining method with a bootstrap value of 1000. The numbers at each node represent neighbor-joining distances. The red block indicates the BmβGRP4.
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.
Phylogenetic relationships of Ichneumonid wasps. The topology is inferred from amino acid sequences of 13 protein-coding genes in mitochondrial genomes (AA matrices) using BI. The numbers close to the nodes separated by "/" represent the ultrafast bootstrap values and Bayesian posterior probabilities in different analysis, respectively. Additionally, "*" represents the full support and "-" represents nonsupport of the corresponding node by that analysis. The order of value for the corresponding analysis is AA by BI/AA by ML/NU by BI/NU by BI. The large "*" alone stands with full supports of all analyses. The triangles on the left confirm some conflicts nodes on the tree through likelihood mapping analysis. The relationship on the top of each triangle is supported by the colored area.
Ichneumonidae is one of the largest families of insects with a mega-diversity of specialized morphological and biological characteristics. We newly sequenced 92 mitochondrial genomes of ichneumonid wasps and found that they have a conserved base composition and a lower evolutionary rate than that of other families of parasitic Hymenoptera. There are 38 types of gene order in the ichneumonid mitochondrial genome, with 30 novel types identified in 104 ichneumonids. We also found that the rearrangement events occur more frequently in Ophioniformes than in Ichneumoniformes and Pimpliformes. Furthermore, the higher Ophioniformes and their relative lineages shared the transposition of trnL2 to trnI-trnQ-trnM tRNA cluster. We confirmed five higher-level groupings of Ichneumonidae: Brachycyrtiformes, Ichneumoniformes, Ophioniformes, Pimpliformes and Xoridiformes. Two formerly unplaced subfamilies, Eucerotinae and Microleptinae, were placed in Brachycyrtiformes and Ichneumoniformes, respectively. The results will improve our understanding of the diversity and evolution of Ichneumonidae.
Concentration-responses of different genetic populations of H. zea to Cry1A.105 in diet overlay bioassays.
Stone's (D) and effective dominance (D ML ) levels of Cry1A.105 resistance in H. zea.
Testing for monogenic inheritance of Cry1A.105 resistance in Cry1A.105-RR'.
Cry1A.105 is a bioengineered Bacillus thuringiensis (Bt) insecticidal protein consisting of three domains derived from Cry1Ac, Cry1Ab, and Cry1F. It is one of the two pyramided Bt toxins expressed in the MON 89034 event, a commonly planted Bt maize trait in the Americas. Recent studies have documented that field resistance of the corn earworm, Helicoverpa zea (Boddie), to the Cry1A.105 toxin in maize plants has become widespread in the United States. To investigate the inheritance of resistance to Cry1A.105 in H. zea, two independent tests, each with various genetic crosses among susceptible and Cry1A.105-resistant populations, were performed. The responses of these susceptible, resistant, F1, F2, and backcrossed insect populations to Cry1A.105 were assayed using a diet overlay method. The bioassays showed that the resistance to Cry1A.105 in H. zea was inherited as a single, autosomal, nonrecessive gene. The nonrecessive nature of the resistance could be an important factor contributing to the widespread resistance of maize hybrids containing Cry1A.105 in the United States. The results indicate that resistance management strategies for Bt crops need to be refined to ensure that they are effective in delaying resistance evolution for nonrecessive resistance (nonhigh dose).
Toxicity of 11 Bt toxins against the black cutworm (BCW), Agrotis ipsilon (Hufnagel).
Black cutworm (BCW), Agrotis ipsilon (Hufnagel), is an occasional pest of maize that can cause considerable economic loss and injury to corn seedlings. This research mainly assessed the susceptibility of BCW neonates to 11 Bt toxins (Cry1Ab, Cry1Ac, Cry1Ah, Cry1F, Cry1Ie, Cry1B, Cry2Aa, Vip3_ch1, Vip3_ch4, Vip3Ca2, Vip3Aa19) by exposing neonates to an artificial diet containing Bt toxins and evaluated the efficacy of three transgenic maize events (C008, C009, C010) expressing Vip3Aa19 toxin against BCW. The toxin-diet bioassay data indicated that Vip3Aa19 protein (LC50 = 0.43 μg/g) was the most active against BCW. Chimeric protein Vip3_ch1 (LC50 = 5.53 μg/g), Cry1F (LC50 = 83.62 μg/g) and Cry1Ac (LC50 = 184.77 μg/g) were less toxic. BCW was very tolerant to the other Bt toxins tested, with LC50 values more than 200 μg/g. Greenhouse studies were conducted with artificial infestations at the whorl stage by placing second-instar BCW larvae into whorl leaf and the fourth-instar larvae at the base of maize seedings. These results suggest that these transgenic maize events expressing Vip3Aa19 can provide effective control for BCW.
Circular map of the mitochondrial genome of Shaddai sp., Sobrala sp., Dikraneura (Dikraneura) zlata, Dikraneurini sp., Alebroides salici and Empoasca serrata.
Relative synonymous codon usage (RSCU) in the mitogenomes of Shaddai sp., Sobrala sp., Alebroides salicis, Empoasca serrata, Empoasca onukii and Dikraneura (D.) zlata.
Predicted secondary cloverleaf structure for the tRNAs of Alebroides salicis. Predicted secondary cloverleaf structure for the tRNAs of Alebroides salicis.
Classification and origins of the mitogenomic sequences used in this study.
To explore the characteristics of mitogenomes and discuss the phylogenetic relationships and molecular evolution of the six tribes within Typhlocybinae, 11 complete mitogenomes are newly sequenced and comparatively analyzed. In all of these complete mitogenomes, the number and order of the genes are highly conserved in overall organization. The PCGs initiate with ATN/TTG/GTG and terminate with TAA/TAG/T. Almost all tRNAs are folded into the typical clover-leaf secondary structure. The control region is always variable in length and in numbers of multiple tandem repeat units. The atp8 and nad2 exhibits the highest evolution rate among all the PCGs. Phylogenetic analyses based on whole mitogenome sequences, with three different datasets, using both maximum likelihood and Bayesian methods, indicate the monophyly of Typhlocybinae and its inner tribes, respectively, except for Typhlocybini and Zyginellini that are paraphyletic. Finally, we confirm that Erythroneurini is a subtribe of Dikraneurini.
Biodiversity of Coccinellidae in some regions of the Caucasus.
Studies of the history of regional insect fauna are important for understanding the changes in ecosystems. We analyzed the dynamics of ladybird fauna at the main sea resort of Russia over a period of 120 years to determine the following: (1) what species disappeared and what species appeared during landscape transformation; (2) what alien species introduced for pest control have been observed to date; and (3) whether the establishment of the global invader Harmonia axyridis (Coccinellidae) caused the extinction of some ladybird species in the Caucasus. We examined specimens collected by us and 54 other collectors including specimens from old museum collections and detected 62 species, 50 of which were collected in recent years (2011–2020). Landscape transformation and recreational use have caused not a decrease but an increase in ladybird biodiversity. Twenty-nine of 34 species recorded before 1930 have been observed in the region to date. Twenty-three other species have spontaneously spread to the region between 1930 and 2020 because of the creation of suitable anthropogenic habitats or because of unintentional introduction. Rodolia cardinalis, Cryptolaemus montrouzieri, Lindorus lophanthae, and Serangium montazerii were released for pest control, and they occur in gardens and streets but not in natural habitats. Harmonia axyridis, which appeared approximately 10 years ago, is abundant in urban and natural habitats, but there is no evidence that it caused the elimination of any ladybird species.
Aphidlions are larvae of certain lacewings (Neuroptera), and more precisely larvae of the groups Chrysopidae, green lacewings, and Hemerobiidae, brown lacewings. The name ‘aphidlion’ originates from their ecological function as specialised predators of aphids. Accordingly, they also play an economic role as biological pest control. Aphidlions have, mostly, elongated spindle-shaped bodies, and similarly to most lacewing larvae they are equipped with a pair of venom-injecting stylets. Fossils interpreted as aphidlions are known to be preserved in amber from the Cretaceous (130 and 100 million years ago), the Eocene (about 35 million years ago) and the Miocene (about 15 million years ago) ages. In this study, new aphidlion-like larvae are reported from Cretaceous amber from Myanmar (about 100 million years old) and Eocene Baltic amber. The shapes of head and stylets were compared between the different time slices. With the newly described fossils and specimens from the literature, a total of 361 specimens could be included in the analysis: 70 specimens from the Cretaceous, 5 from the Eocene, 3 from the Miocene, 188 extant larvae of Chrysopidae, and 95 extant larvae of Hemerobiidae. The results indicate that the diversity of head shapes remains largely unchanged over time, yet there is a certain increase in the diversity of head shapes in the larvae of Hemerobiidae. In certain other groups of Neuroptera, a distinct decrease in the diversity of head shapes in larval stages was observed.
(a) The expression of miR-14 in queens after treatment and (b) the effect of its expression on the egg number of queens. NC, queens without treatment. miR-14 stable NC, miR-14 stable nonsense sequence control. Inhibitor NC, inhibitor nonsense sequence control. * means p < 0.05, ** means p < 0.01.
Effect of siRNAs and overexpressed vector on the expression of EcR. (a) Effect of siRNA on the expression of EcR. (b) Effect of overexpressed vector on the expression of EcR. * means p < 0.05, ** means p < 0.01.
(a) The expression of EcR in queens after treatment and (b) the effect of its expression on the egg number of queens. EcR siRNA NC, EcR siRNA nonsense sequence control. Normal control, queens without treatment. * means p < 0.05, ** means p < 0.01.
(a) Sequences of the interaction sites between miR-14 and EcR 3 UTR and (b) co-transfection of psiCHECK2-EcR 3 UTR resulted in dramatic suppression of the luciferase activity. a. Blue line indicates interaction sites, and asterisks indicate mutated site. Nucleotides of interaction sites and mutated sites are shown in red. Grey shaded areas indicate canonical 7mer "seed" region that aligns with the target site, the vertical lines indicate contiguous Watson-Crick pairing. b. A normalized firefly/renilla luciferase value was plotted with ±SD. ** means p < 0.01.
Effect of miR-14 on mRNA and protein level of EcR. (a) Effect of miR-14 on mRNA level of EcR. (b) Western blot result. (c) Effect of miR-14 on protein level of EcR at 24 h and 48 h after treatment. * denoting p < 0.05, ** denoting p < 0.01. Data are from three replicates. β-actin was used as the reference protein.
Honeybees (Apis mellifera) are important pollinators and are commonly used for honey production. The oviposition behavior in honeybees is complex and errors in oviposition could affect the development of the bee colony. Recent studies reported that RNA-RNA cross-talk played a critical role in several biological processes, including reproduction. Ecdysone receptor (EcR) and miR-14 were previously reported to play important roles in egg-laying. Moreover, EcR was predicted to be the target gene of miR-14 and may form miR-14-EcR cross-talk. In this study, knocking down and overexpression of miR-14 and EcR in queen model were implemented. The effect of RNA expression of miR-14 and EcR on the number of eggs laid by honeybee queens were analyzed. Further, luciferase assay was used to confirm the target relation between miR-14 and 3'UTR of EcR. The results showed that the expression of miR-14 and EcR was associated with the number of eggs laid by queens. In specific, inhibition of miR-14 expression enhanced the number of eggs laid, while overexpression of EcR enhanced the number of eggs laid. Lastly, we determined that miR-14 directly targets the mRNA of EcR. These findings suggest that the cross-talk of miR-14-EcR plays an important role in the number of eggs laid by honeybee queens.
In this study, 50 species of the leafhopper subgenus Eurhadina (Singhardina) Mahmood from China are reviewed based on comparative morphological characteristics, including 14 new species: Eurhadina (Singhardina) amacularis, E. (S.) extensa, E. (S.) flaviscutella, E. (S.) foliiformis, E. (S.) galacta, E. (S.) gracilifurca, E. (S.) lata, E. (S.) parilintanonica, E. (S.) quadrimacularis, E. (S.) recta, E. (S.) scalesa, E. (S.) scamba, E. (S.) scandens and E. (S.) uprotrusa sp. nov. Four additional species E. (S.) fasciata, E. (S.) jarrayi, E. (S.) prima and E. (S.) zadyma are recorded from China for the first time. Two new synonymies are proposed. Eurhadina (Singhardina) flavescens Huang et Zhang, 1999 syn. nov. is synonymized with Eurhadina wuyiana Yang et Li, 1991 and Eurhadina rubromia Cai et Kuoh, 1993 syn. nov. is synonymized with Eurhadina (Singhardina) biavis Yang et Li, 1991. A key to all Chinese Singhardina species is also provided.
Mean percentages (±SD) of untreated and 150 Gy-treated Lobesia botrana males succeeding in flying to virgin untreated and 150 Gy-treated calling females in the calling female compartment in a flight assessment cage during 4 successive days. The mean percentages (±SD) of males that failed to fly into the female compartment is consigned in the column farthest to the right.
Mean percentage of egg hatch (±SD) and percentage of twice-mated females in Lobesia botrana females mated sequentially with 150 Gy-treated and untreated males.
The sterile insect technique/inherited sterility (SIT/IS) has been suggested as an eco-friendly control tactic for area-wide integrated pest management approaches in order to control the European grapevine moth, Lobesia botrana. This study assessed the effects of an irradiation dose of 150 Gy administered to newly emerged female moths on their egg laying behaviour and mating ability at different ages at mating. Moreover, the effects of multiple mating on the mating ability of treated females, pattern of sperm precedence in twice-mated females and the flight response of treated and untreated males to treated and untreated calling females were also investigated. Females treated with 150 Gy initiated calling in a way similar to untreated females. When treated females were paired with untreated males, the mean number of eggs oviposited per female during 6 days was reduced (59.6 and 82.8 eggs/female, respectively), as was their mating ability and multiple mating compared with untreated females. The proportion of offspring fertilized by the second of the two males to mate with the female or last-male sperm precedence (P2 value) constituted 97% of the eggs, suggesting that the second male mate fathered the most offspring. The outcome of this work could be viewed as an integrated approach for improving effectiveness and enabling successful implementation of a SIT/IS program against L. botrana.
The spotted alfalfa aphid (Therioaphis trifolii (Monell)) is a known destructive pest that can significantly reduce alfalfa yields. Two differentially up-regulated alfalfa trypsin inhibitors ‘Msti-94’ and ‘Msti-16’ in transcriptome were verified in terms of their mRNA levels using RT-qPCR. The prokaryotic expression vector was constructed and its biological functions, including phenotypic and physiological responses, were verified through feeding spotted alfalfa aphids with active recombinant protein mixed with an artificial diet. Gene clone and gene prokaryotic expression confirmed that Msti-94 had a size of 651 bp, encoded 216 amino acids with a predicted protein weight of 23.5 kDa, and a pI value of 6.91. Similarly, the size of Msti-16 was 612 bp, encoded 203 amino acids, and had a predicted protein weight of 22.2 kDa with a pI value of 9.06. We concluded that both Msti-94 and Msti-16 acted as a stomach poison with survival rates reduced to 21.7% and 18.3%, respectively, as compared to the control, where the survival rate was significantly (p < 0.05) higher (60.0%). Aphid reproduction rates were significantly reduced, after 72 h of feeding, in both the Msti-94 and Msti-16 treatments compared to the controls. A concentration of 800 μg/mL (0.8 mg/mL) of recombinant protein and 5000 μg/mL (5 mg/mL) of recombinant expressing bacteria that inhibits the total protease, which ultimately disrupted the activity of trypsin, chymotrypsin, and aminopeptidase.
Sampling locations and GPS information for Depressaria depressana collected from wild car- rot (Daucus carota) for this study.
Reconstruction of the establishment of D. radiella across the USA and Canada with the use of literature and museum records.
Depressaria depressana, the purple carrot seed moth, is a Eurasian species first reported in North America in 2008 and currently undergoing range expansion. This invasion follows that of its Eurasion congener Depressaria radiella (parsnip webworm), first documented in North America 160 years ago. Unlike D. depressana, which utilizes hostplants across multiple tribes of Apiaceae, Depressaria radiella is a “superspecialist” effectively restricted in its native and non-indigenous ranges to two closely related apiaceous genera. We investigated the genetic structure of D. depressana populations across latitudinal and longitudinal gradients in the eastern United States by constructing COI haplotype networks and then comparing these with haplotype networks constructed from available COI sequence data from contemporary European D. depressana populations and from European and North American D. radiella populations. Haplotype data revealed higher genetic diversity in D. depressana, indicating high dispersal capacity, multiple introductions, and/or a genetically diverse founding population. Museum and literature records of D. radiella date back to 1862 and indicate that range expansion to the West Coast required more than 50 years. Higher levels of genetic diversity observed in D. depressana compared to its congener may indicate a greater propensity for dispersal, colonization and establishment in its non-indigenous range.
Genbank sequences used in phylogenetic analysis. Accession numbers are provided. Dashed lines indicate a species was used in only one of the phylogenies. Double asterisks ** indicate the outgroup. 
Armored scale insects pose a serious threat to habitat conservation across the globe because they include some of the most potent invasive species in the world. They are such a serious concern because their basic morphology, small size, and polyphagous feeding habits often allow them to exist undetected by growers and quarantine experts. In order to provide a potential solution to the problem, we have attempted to elucidate the effectiveness of molecular identification techniques using ribosomal 28s and endosymbiotic 16s rRNA. Sequence data was obtained from many field-collected insects to test the feasibility of identification techniques. A protocol for quick species determination based on sequence data is provided.
Responses of female Sitodiplosis mosellana adults to synthetic odors derived from wheat volatiles. All tested compounds subjected to Y-tube olfactometer assays were diluted with liquid paraffin to a final concentration of 20 μg/μL and liquid paraffin was used as the solvent control. N marks the number of individuals that made a choice out of 60 tested insects, and the insects that did not make a choice are excluded from the statistical analysis. Asterisk denotes significant difference (p < 0.05), and n.s. indicates no significant difference by chi-square tests.
Primers used in cloning and expression of Sitodiplosis mosellana odorant-binding proteins 12 and 17 (SmosOBP12 and SmosOBP17).
Binding affinities of two SmosOBPs to wheat volatile compounds in fluorescence binding assays.
The wheat blossom midge Sitodiplosis mosellana, one of the most disastrous wheat pests, depends highly on olfactory cues to track suitable plants. To better understand the olfactory recognition mechanisms involved in host selection, in the present study we cloned two S. mosellana adult antenna-specific odorant binding protein (OBP) genes, SmosOBP12 and SmosOBP17, and evaluated bacterially expressed recombinant proteins for their selectivity and sensitivity for host wheat volatiles using the fluorescence-based ligand binding assay. The results showed that both SmosOBPs effectively bound alcohol, ester, ketone, and terpenoid compounds. Particularly, SmosOBP12 had significantly higher affinities (Ki < 10.5 μM) than SmosOBP17 (Ki2 > 0.1 μM) to 3-hexanol, 1-octen-3-ol, D-panthenol, 3-carene, (Z)-3-hexenylacetate, hexyl acetate, methyl salicylate, heptyl acetate, and ethyl heptanoate. Consistently, S. mosellana females were attracted to all these chemicals in a behavioral assay using Y-tube olfactometer. SmosOBP12 also bound aldehyde, but neither bound alkanes. Notably, SmosOBP12 exhibited strong affinity to ocimene (Ki = 8.2 μM) that repelled S. mosellana. SmosOBP17, however, was insensitive to this compound. Taken together, our results indicate that SmosOBP12 may play a greater role than SmosOBP17 in perceiving these biologically active plant volatiles.
Two pig carcasses on the pig platform, attached to the instrument platform, ready to be deployed. A digital webcam was positioned above the carcasses. Frame, cage, and trays designed by Chris Sundstom and instrument platform designed by Paul Macoun, ONC, VENUS observatory.
Daily average dissolved oxygen levels (a) and temperature (b) over the duration of the experiments at a depth of 170 m in the Strait of Georgia in spring and fall (Ocean Network Canada's VENUS observatory).
The taphonomy of carcasses submerged in the ocean is little understood, yet it is extremely important ecologically and forensically. The objectives of this study were to determine the fate of pig carcasses as human proxies in the Strait of Georgia at 170 m in spring and fall. Using Ocean Networks Canada’s Victoria Experimental Network Underseas (VENUS) observatory, two carcasses per season were placed under a cabled platform hosting a webcam and instruments measuring water chemistry. Two minutes of video were recorded every 15 min. In spring, Lyssianassidae amphipods and Pandalus platyceros were immediately attracted and fed on the carcasses, the amphipods removed the bulk of the soft tissue from the inside whilst the shrimp shredded the skin and tissue. The carcasses were skeletonized on Days 8 and 10. In fall, Metacarcinus magister was the major scavenger, removing most of the soft tissue from one carcass. Amphipods did not arrive in large numbers until Day 15, when they skeletonized the scavenged carcass by Day 22 and the less scavenged carcass by Day 24. Amphipods remained for some days after skeletonization. This skeletonization was very different from previous experiments at different depths and habitats. Such data are very valuable for predicting preservation, planning recoveries, and managing family expectations.
Predicted changes in the distribution areas of suitable habitat for Callosobruchus chinensis in China between two adjacent time periods. "-1" represents the expansion areas, which is the green region in figures; "0" represents the areas where the species did not exist, which is the white region in figures; "1" represents the areas where the distribution had not changed, which is the yellow region in figures; "2" represents the areas where the distribution was decreased, and this is the gray region in figures.
MJ analysis of the network of Callosobruchus chinensis based on concatenated mitochondrial haplotypes. The circles represent different haplotypes with their proportional to the number of individuals. The colors represent different populations. The short line segments indicate mutated positions between haplotypes.
Primers' sequences for amplification of four mitochondrial genes (COI, COII, Cyt-b and 12S rRNA) for Callosobruchus chinensis.
Nucleotide polymorphisms among 2803 bp of mtDNA sequence from Callosobruchus chinensis beetles in population samples across China.
Callosobruchus chinensis (Coleoptera Bruchidae), is a pest of different varieties of legumes. In this paper, a phylogeographical analysis of C. chinensis was conducted to provide knowledge for the prevention and control of C. chinensis. A total of 224 concatenated mitochondrial sequences were obtained from 273 individuals. Suitable habitat shifts were predicted by the distribution modelling (SDM). Phylogeny, genetic structure and population demographic history were analyzed using multiple software. Finally, the least-cost path (LCP) method was used to identify possible dispersal corridors and genetic connectivity. The SDM results suggested that the distribution of C. chinensis experienced expansion and contraction with changing climate. Spatial distribution of mtDNA haplotypes showed there was partial continuity among different geographical populations of C. chinensis, except for the Hohhot (Inner Mongolia) population. Bayesian skyline plots showed that the population had a recent expansion during 0.0125 Ma and 0.025 Ma. The expansion and divergent events were traced back to Quaternary glaciations. The LCP method confirmed that there were no clear dispersal routes. Our findings indicated that climatic cycles of the Pleistocene glaciations, unsuitable climate and geographic isolation played important roles in the genetic differentiation of C. chinensis. Human activities weaken the genetic differentiation between populations. With the change in climate, the suitable areas of C. chinensis will disperse greatly in the future.
Collecting locality of specimens used in this study.
Median-joining haplotype network for 16S rDNA sequences. Different colours are related to subspecies, recognized according to Sturm [3]. The number of individuals with the same haplotype is provided below the haplotype name. Hatch marks along edges represent the number of mutations between nodes. (Hap 17 for 16S rDNA and Hap 25 for cytochrome b), differing from the most common haplotypes only by one and two mutations. The phylogenetic analysis also indicated the lack of sequences distinctiveness of the British specimens of P. machaon.
Median-joining haplotype network for cytochrome b sequences. Different colours are related to subspecies, recognized according to Sturm [3]. The number of individuals with the same haplotype is provided below the haplotype name. Hatch marks along edges represent the number of mutations between nodes.
The maximum-likelihood tree derived from a combination of 16S rDNA and cytochrome b sequences. Bootstrap values over 70% are shown next to the branches. The scale bar shows the number of substitutions per site. The individuals on the tree are related to subspecies according to approach of Eller [1] (first column), Seyer [2,7-9] (second column) and Sturm [3] (third column).
Standard genetic indices and average pairwise genetic distance of 16S rDNA marker.
The Old World swallowtail Papilio machaon Linnaeus, 1758 is one of the most well-known and most characteristic members of the family Papilionidae. Over the past two centuries, the butterfly has been the subject of many studies. P. machaon is characterised by a tendency to change the wing colour pattern. In turn, due to the great interest of collectors and amateur entomologists, these studies have been converted into the description of over 100 colour forms, aberrations and subspecies. In this study, mitochondrial DNA (mtDNA), 16S rDNA and cytochrome b sequences were used to examine the correlation between the intraspecific classification and genetic structure of P. machaon. The study used 87 specimens from 59 different localities covering the geographic distribution of this species in the palaearctic. The phylogenetic relationships within and between the Old World swallowtail subspecies showed that the intraspecific classification proposed by various authors does not correlate with the variability in mitochondrial DNA sequences. In addition, populations occurring at the species distribution borders in the palaearctic region (i.e., Japan, Kamchatka, Morocco and Sakhalin) are genetically distinct from other species.
Studies of bacterial communities can reveal the evolutionary significance of symbiotic interactions between hosts and their associated bacteria, as well as identify environmental factors that may influence host biology. Atta sexdens is an ant species native to Brazil that can act as an agricultural pest due to its intense behavior of cutting plants. Despite being extensively studied, certain aspects of the general biology of this species remain unclear, such as the evolutionary implications of the symbiotic relationships it forms with bacteria. Using high-throughput amplicon sequencing of 16S rRNA genes, we compared for the first time the bacterial community of A. sexdens (whole ant workers) populations according to the habitat (natural versus agricultural) and geographical location. Our results revealed that the bacterial community associated with A. sexdens is mainly influenced by the geographical location, and secondarily by the differences in habitat. Also, the bacterial community associated with citrus differed significantly from the other communities due to the presence of Tsukamurella. In conclusion, our study suggests that environmental shifts may influence the bacterial diversity found in A. sexdens.
Generalized linear models (deviance analysis, type III) for the number and duration visits recorded on watermelon triploid flowers. The independent variables were release densi 30 and 45 individuals/m 2 ) and flower type (staminate versus pistillate).
Number of pollen grains adhered (mean ± SE), minimum and maximum values between parentheses, on the stigma of triploid watermelon flowers for each treatment (release density: HD, MD and LD), in both years of the trial.
Number of fruits per plant, fruit weight, and yield in the enclosures, for both waterme cultivars. Years 2020 and 2021 for the three treatments (release density: HD, MD and LD).
Watermelon (Citrullus lanatus) is an important crop worldwide. Pollination of this crop is carried out by insects, with honey bees (Apis spp.) and bumble bees (Bombus spp.) as the most used in greenhouse production. Nevertheless, due to the extreme conditions in closed enclosures, these hymenopterans suffer management and behavior problems leading to insufficient pollination. The effectiveness of three release densities (15, 30, and 45 individuals/m2) of Eristalinus aeneus was compared in diploid- and triploid-associated watermelon varieties under protected cultivation. Floral visits, pollen–pistil interaction after pollen transport, yield, and fruit quality were evaluated. The number of floral visits increased with release density in both pistillate and staminate flowers. No significant differences were observed, however, among release densities or between flower types in the duration of the visits. Floral preferences were not found in the behavior of E. aeneus in watermelon. High and medium release densities increased pollen deposition onto the stigma, and consequently the yield of the triploid variety compared to low release density, by 23.8 to 41.8% in 2020 and by 36.3 to 46.7% in 2021. The results of this trial demonstrate the potential of E. aeneus as a managed pollinator in protected cultivation of triploid watermelon.
Protein contents (%) of defatted flour, concentrates and isolates of Brachytrupes membranaceus and Macrotermes subhyalinus.
Amino acid composition (g/100 g protein) of defatted flour, protein concentrate, and isolate of Brachytrupes membranaceus.
Amino acid composition (g/100 g protein) of defatted flour, protein concentrate, and isolate of Macrotermes subhyalinus.
Digestibility (%) of defatted flour, concentrates, and isolates of Brachytrupes membranaceus and Macrotermes subhyalinus.
Brachytrupes membranaceus and Macrotermes subhyalinus are edible insects in Burkina Faso. Our research aimed to evaluate the nutritional composition and functional properties of the defatted flours, protein concentrates, and isolates of Brachytrupes membranaceus and Macrotermes subhyalinus. Proximate and mineral composition were determined according to AOAC methods. The amino acid and fatty acid composition were determined by high-performance liquid chromatography (HPLC) and gas chromatography, respectively. The protein concentrates and isolates were obtained by solubilization, precipitation, and lyophilization. Macrotermes subhyalinus showed the highest protein (45.75 g/100 g), iron (11.76 mg/100 g), and zinc (13.18 mg/100 g) contents. The highest isoleucine and lysine contents, the best fat absorption (10.87 g/g), and foaming capacities (49.60%) were obtained with the isolate of Brachytrupes membranaceus. Consumption of Macrotermes subhyalinus could be used to fight or correct iron and zinc deficiencies. Macrotermes subhyalinus was a source of macronutrients and micronutrients, while the protein concentrates and isolates of Brachytrupes membranaceus were endowed with functional properties (fat absorption and foaming capacities).
Means and 95% confidence intervals of activity (a), courtship (b), boldness (c), and body size (d) of Calopteryx splendens between six populations: 1-2 (southern region), 3-4 (central region), and 5-6 (northern region).
Sampling sites (six populations) and sample sizes of Calopteryx splendens males in three regions: southern, central and northern.
Description of behavioural traits measured in three behavioural axes of Calopteryx splendens from southern, central and northern region.
Post hoc pairwise comparisons of Calopteryx splendens male behaviours and body size between three regions: southern (S), central (C) and northern (N).
Behavioural variation is important for evolutionary and ecological processes, but can also be useful when predicting consequences of climate change and effects on species ranges. Latitudinal differences in behaviour have received relatively limited research interest when compared to morphological, life history and physiological traits. This study examined differences in expression of three behavioural axes: activity, courtship and boldness, and their correlations, along a European latitudinal gradient spanning ca. 1500 km. The study organism was the temperate damselfly Calopteryx splendens (Harris). We predicted that the expression of both behavioural traits and behavioural syndromes would be positively correlated to latitude, with the lowest values in the southern populations, followed by central and the highest in the north, because animals usually compensate behaviourally for increasing time constraints and declining environmental conditions. We found that behavioural expression varied along the latitudinal cline, although not always in the predicted direction. Activity was the only behaviour that followed our prediction and gradually increased northward. Whereas no south-to-north gradient was seen in any of the behavioural syndromes. The results, particularly for activity, suggest that climatic differences across latitudes change behavioural profiles. However, for other traits such as courtship and boldness, local factors might invoke stronger selection pressures, disrupting the predicted latitudinal pattern.
Red palm weevil (Rhynchophorus ferrugineus) (a) aggregating male and female adults (b) male mouthpart with chemosensory hairs (c) female mouthpart exhibiting sexual dimorphism (dg) RPW farming (d) a local farm with a big pile of minced sago bark for rearing beetles (e) artificial nests for breeding beetles (f) larvae for sale as an exotic snack (g) a farmer is preparing adult beetles for breeding.
Antennal and mouthpart transcriptome data (a) Data distribution of the Blast2GO result (b) Gene ontology distribution by level (top 20) (c) Top-hit species distribution (d) Top 50 most expressed genes in the four transcriptomes. Genes are grouped into main classes. Genes that are not classified into a group are included in 'others.'
Phylogenetic tree of the red palm weevil and other pest beetle gustatory receptors. Due to its large size, tree was divided into three parts: (a) bottom, (b) middle, and (c) top. A tree was reconstructed from protein sequences using the maximum likelihood approach by PhyML3.0. The carbon dioxide, sugar, and fructose receptor clades were highlighted. Tree was rooted at the CO2 receptor clade. Branch supports were assessed using approximate likelihood-ratio test (aLRT); a value above 0.9 is shown with an orange dot on the node (Rfer = Rhynchophorus ferrugineus, Tcas = Tribolium castaneum, Dpon = Dendroctonus ponderosae, Ldec = Leptinotarsa decemlineata).
Chromosome mapping of the Rhynchophorus ferrugineus gustatory receptor genes (a) distribution of RferGr genes on the ten chromosomes (b) inverted duplication of the sugar receptor Chromosome mapping of the Rhynchophorus ferrugineus gustatory receptor genes (a) distribution of RferGr genes on the ten chromosomes (b) inverted duplication of the sugar receptor genes (Color code for Gr genes: blue = candidate c receptor, red = candidate sugar receptor, black = candidate bitter receptor).
The red palm weevil (Rhynchophorus ferrugineus) is a highly destructive pest of oil palm, date, and coconut in many parts of Asia, Europe, and Africa. The Food and Agriculture Organization of the United Nations has called for international collaboration to develop a multidisciplinary strategy to control this invasive pest. Previous research focused on the molecular basis of chemoreception in this species, particularly olfaction, to develop biosensors for early detection and more effective bait traps for mass trapping. However, the molecular basis of gustation, which plays an essential role in discriminating food and egg-laying sites and chemical communication in this species, is limited because its complete gustatory receptor gene family still has not been characterized. We manually annotated the gene family from the recently available genome and transcriptome data and reported 50 gustatory receptor genes encoding 65 gustatory receptors, including 7 carbon dioxide, 9 sugar, and 49 bitter receptors. This study provides a platform for future functional analysis and comparative chemosensory study. A better understanding of gustation will improve our understanding of this species’ complex chemoreception, which is an important step toward developing more effective control methods.
The stinging nettle caterpillar, Parasa lepida. (A) Caterpillars feeding on coconut leaves, (B) dermatitis symptom on a hand after exposure to caterpillar, (C) adult stage, and (D) final instar caterpillar with urticating hairs on its back (scale bar = 1 cm).
Flow chart of the P. lepida toxin gene identification pipeline: (A) transcriptome analysis-green box = RNA isolation and sequencing, blue box = transcriptome analysis; (B) toxin gene identification pipeline: identification based on orthologous relationships (left) and identification of novel toxin genes (right).
(A) Box-and-Whisker plot showing expression level (FPKM) of P. lepida venom genes by subfamily. Boxes show the upper quartile and lower quartile. The line across the box indicates the median. The maximum and minimum values (excluding outliers) are shown above and under the box, respectively. Upper and lower outliers are indicated using red and pink symbols, respectively. (B) The expression level of the top 20 most expressed P. lepida toxin genes.
(A) Protein alignment of novel Knottin-domain protein from P. lepida showing six conserved cysteine residues and putative three disulfide bridges; (B) phylogenetic tree of Knottin-like proteins from P. lepida and other animals: Conotoxin-Conus achatinus (Little frog cone), Antimicrobial Knottin protein-Bemisia tabaci (Silverleaf whitefly), OpicalcinOpistophthalmus carinatus (African yellow leg scorpion), Hainantoxin-Cyriopagopus hainanus (Chinese bird spider), Grammotoxin-Grammostola rosea (Chilean rose tarantula). Branch supports higher than 70% (1000 replications bootstrapping) are highlighted with grey circles.
Information of sequence reads generated by RNA-seq, and Trinity de novo-assembled contigs.
Many animal species can produce venom for defense, predation, and competition. The venom usually contains diverse peptide and protein toxins, including neurotoxins, proteolytic enzymes, protease inhibitors, and allergens. Some drugs for cancer, neurological disorders, and analgesics were developed based on animal toxin structures and functions. Several caterpillar species possess venoms that cause varying effects on humans both locally and systemically. However, toxins from only a few species have been investigated, limiting the full understanding of the Lepidoptera toxin diversity and evolution. We used the RNA-seq technique to identify toxin genes from the stinging nettle caterpillar, Parasa lepida (Cramer, 1799). We constructed a transcriptome from caterpillar urticating hairs and reported 34,968 unique transcripts. Using our toxin gene annotation pipeline, we identified 168 candidate toxin genes, including protease inhibitors, proteolytic enzymes, and allergens. The 21 P. lepida novel Knottin-like peptides, which do not show sequence similarity to any known peptide, have predicted 3D structures similar to tarantula, scorpion, and cone snail neurotoxins. We highlighted the importance of convergent evolution in the Lepidoptera toxin evolution and the possible mechanisms. This study opens a new path to understanding the hidden diversity of Lepidoptera toxins, which could be a fruitful source for developing new drugs.
Image of the wing of Hydrotaea capensis taken with the binocular stereoscope with the measurements made using the TpsDig2 program according to different morphometric geometry studies. AB, in red; Am/Bm, in yellow; 1-2, in dark blue; c1-c2, in blue; r2-r5, in pink.
Length (average ± standard error, in mm) of the different biometric wing measurem H. capensis (F: females, M: males) at 20 °C, 25 °C and 30 °C. No adults emerged at 18 °C. Te temperature, N is the sample size. Different letters indicate statistically significant difference test, p < 0.05).
Length (average ± standard error, in mm) of the different biometric wing measurements of H. capensis (F: females, M: males) at 20 • C, 25 • C and 30 • C. No adults emerged at 18 • C. Temp. is temperature, N is the sample size. Different letters indicate statistically significant differences (LSD test, p < 0.05).
One of the most important and perhaps most used applications of forensic entomology concerns the estimation of the minimum post-mortem interval (minPMI), defined as the time interval between death and the actual finding of a corpse. Some Diptera species are critical in these studies because they are the first ones capable of detecting and finding a corpse and are selectively attracted by its decomposing status. Thus, the knowledge of the micromorphology of their preimaginal stages and of their life cycles within a time frame constitutes solid indicators for estimating the minPMI. Hydrotaea capensis is a Muscidae of forensic interest usually considered as a late colonizer of corpses. It is widely distributed, living mainly in warm regions, and present in a wide variety of habitats. In this study, the H. capensis life cycle was studied at four constant temperatures, 18°, 20°, 25° and 30 °C, by recording the duration of its different developmental stages, including the length reached in each larval stage, as well as some biometric characteristics of the emerged adults. Significant differences were observed in the average time of development of most larval stages, with a longer duration at low temperatures, and in the length of each larval stage depending on the temperature, but, in this case, without a clear pattern. Moreover, significant differences were found in some alar features, pointing to them as a relevant indicator to be considered. The data provided will assist forensic entomologists to make more accurate minPMI estimations in cases where H. capensis is present.
Leaf-rollers and tent caterpillars, the families Torticidae and Lasiocampidae, represent a significant component of the Lepidoptera, and are well-represented in the forest insect pest literature of North America. Two species in particular—spruce budworm (Choristoneura fumiferana (Clem.)) and forest tent caterpillar (Malacosoma disstria Hbn.)—are the most significant pests of the Pinaceae and Salicacae, respectively, in the boreal forest of Canada, each exhibiting periodic outbreaks of tremendous extent. Dispersal is thought to play a critical role in the triggering of population eruptions and in the synchronization of outbreak cycling, but formal studies of dispersal, in particular studies of long-range dispersal by egg-bearing adult females, are rare. Here, it is shown in two independent studies that adult females of both species tend to disperse away from sparse or defoliated forest, and toward intact or undefoliated forest, suggesting that long-range dispersal during an outbreak peak is adaptive to the species and an important factor in their population dynamics, and hence their evolutionary biology.
Habitus of male (a) and female (b) Pterostichus adstrictus from the Kaunertal (Ötztal Alps, Austria).
Median lobe of the aedeagus of Pterostichus adstrictus from the Taschachtal (Ötztal Alps, Austria), (a) right lateral view, (b) left lateral view, scale bar = 1 mm.
Comparison of the aedeagi of Pterostichus adstrictus, Kaunertal/Austria (a), P. oblongopunctatus, Rabachboden/Austria (b) and P. quadrifoveolatus, Lopenik/Czech Republic (c), median lobe in ventral view, scale bar = 1 mm.
Synoptic key for the three European Pterostichus species of the subgenus Bothriopterus. The determination charac- ters are compiled from own new data and the literature [27-29].
The last ice age considerably influenced distribution patterns of extant species of plants and animals, with some of them now inhabiting disjunct areas in the subarctic/arctic and alpine regions. This arctic-alpine distribution is characteristic for many cold-adapted species with a limited dispersal ability and can be found in many invertebrate taxa, including ground beetles. The ground beetle Pterostichus adstrictus Eschscholtz, 1823 of the subgenus Bothriopterus was previously known to have a holarctic-circumpolar distribution, in Europe reaching its southern borders in Wales and southern Scandinavia. Here, we report the first findings of this species from the Austrian Ötztal Alps, representing also the southernmost edge of its currently known distribution, confirmed by the comparison of morphological characters to other Bothriopterus species and DNA barcoding data. Molecular data revealed a separation of the Austrian and Finish specimens with limited to no gene flow at all. Furthermore, we present the first data on habitat preference and seasonality of P. adstrictus in the Austrian Alps.
Cephalotes pusillus digging behavior records. (A-C) Workers unloading the small fragments of wood or sawdust; (D,E) accumulation of sawdust thrown away by the workers above the vegetation under nest. 
Cephalotes pusillus digging behavior records. (A-C) Workers unloading the small fragments of wood or sawdust; (D,E) accumulation of sawdust thrown away by the workers above the vegetation under nest. 
Here we use karyomorphometrical analysis to characterize and evaluate the karyotype of the turtle ant Cephalotes pusillus (Klug, 1824). This is the first representative of this diverse ant genus to be cytogenetically studied. They bear a diploid chromosome set of 44 chromosomes, which, according to the centromeric index, are metacentric, submetacentric, and subtelocentric. This small ant is quite widely distributed in the Neotropics and seems to be well adapted to living in disturbed areas. Here we report the species nesting on dead trunks used to build fences at countryside houses and farms. On these nests, we observed some never reported behavior of C. pusillus: the ants appear to be able to dig by actively removing small fragments of dead wood fiber, hence expanding their nest cavities. It was not thought that Cephalotes species had this ability, given that they nest in preexisting cavities. Our observations are initial remarks that the small plier-like mandibles of C. pusillus may not be a constraint for this species, adding to our knowledge on ant nesting biology.
Jackknife of regularized training gain for the Oedaleus decorus distribution models for the south-eastern part of the West Siberian Plain: (A)-distribution data before 1961 and bioclimatic variables for 1970-2000; (B)-distribution data from 1961 until 2021 and bioclimatic variables for 1970-2000; (C)-distribution data from 1961 until 2021 and forecasts of bioclimatic variables from the climatic model CNRM-ESM2-1; (D)-distribution data from 1961 until 2021 and forecasts of bioclimatic variables from the climatic model MIROC6.
Predictive contributions for two periods (until 1960 and 1961-2021) and for two climatic models for 2021-2040.
Dynamics of the abundance (ind. per hour) of Oedaleus decorus relative to the abundance of the Italian locust in the Kulunda steppe.
Dynamics of the average density (ind. per m 2 ± s.e.) of Oedaleus decorus relative to the average density of the Italian locust in the Kulunda steppe.
Oedaleus decorus is a widely distributed acridid over the Eurasian semi-arid territories, from the Atlantic coast to the Pacific coast. In many semi-arid territories, O. decorus was and is the most important pest, but in the south-eastern part of West Siberian Plain, it was not considered a pest until the 1960s. We compared two sets of data on the acridid distribution in the region: before 1960 and from 1961 until 2021. Until the 1960s, the species occurred mainly in the southern steppes. Since the 1960s, its distribution changed significantly. Nowadays, it occupies almost all local steppes and the southern part of the forest-steppes and can be also found on the eastern side of the Ob River. These shifts may be explained by both climatic changes and changes in human activities. During upsurges the densities of O. decorus were often more than one to two adults per square meter. It is often abundant in the same habitats and in the same periods as the Italian locust (Calliptamus italicus)—one of the most important acridid pests. This means during joint outbreaks these two species can simultaneously damage almost all spectrum of plants.
Male genitalia of Caloptilia. (a). C. purpureus (gen. slide no. HNUSEL-5592); (b). C. acericola (gen. slide no. HNUSEL-5571); (c). C. celtidis (gen. slide no. HNUSEL-5561); (d). C. dentata (gen. slide no. HNUSEL-5549); (e). C. kadsurae (gen. slide no. HNUSEL-5593); (f). C. recitata (gen. slide no. HNUSEL-5544); (g). C. soyella (gen. slide no. HNUSEL-5556, 5557) (Scale bars: a-g = 5 mm).
Female genitalia of Caloptilia. (a). C. purpureus (gen. slide no. HNUSEL-5591); (b). C. koreana (gen. slide no. HNUSEL-5575); (c). C. xanthos (gen. slide no. HNUSEL-5602). (Scale bars: a-e = 5 mm).
Adults of Caloptilia. (a). C. acericola (gen. slide no. HNUSEL-5577); (b). C. celtidis (gen. slide no. HNUSEL-5613); (c). C. dentata (gen. slide no. HNUSEL-5589); (d). C. kadsurae (gen. slide no. HNUSEL-5593); (e). C. monticola (gen. slide no. HNU-SEL-5587); (f). C. recitata (gen. slide no. HNUSEL-5577); (g). C. soyella (gen. slide no. HNUSEL-5557).
In this study, 29 species of Caloptilia Hübner, 1825, belonging to the family Gracillariidae, were recognized in Korea. Among these, three species, i.e., C. purpureus sp. nov., C. koreana sp. nov., and C. xanthos sp. nov., are described as new to science. In addition, seven species of this genus are reported for the first time in Korea. All known species were enumerated, based on their available information. Adult specimens and genitalia of the new and newly recorded species were examined and described using all available information.
The hoverfly Sphaerophoria rueppellii is currently one of the most effective predators commercially available for aphid pest control. However, knowledge of the reproductive system of males and females of this syrphid is limited. The present article aims to report how changes in the temperature and photoperiod may affect development of the gonads (ovaries and testes), oviposition, and fecundity during the lifespan of S. rueppellii. Four environmental conditions (14L:10D, T: 20 ± 1 °C; 12L:12D, T: 20 ± 1 °C; 14L:10D, T: 25 ± 1 °C; and 12L:12D, T: 25 ± 1 °C) were used to determine oviposition, hatching percentage, and lifespan during a period of 30 days after the adult emergence. The maturation of the ovaries was done under three treatments (barley leaves with aphids always available; barley leaves two days per week with aphids available; no barley leaves available), and in the same environmental conditions noted above. Males at 14L:10D, 20 ± 1 °C; and 14L:10D, 25 ± 1 °C; were used to analyze and study the maturation of the testes. Females at 14L:10D; T: 25 ± 1 °C showed a significant difference in oviposition, percentage of hatching, and rate of eggs. A detailed description of the male and female gonads was undertaken, and it was determined that the conditions in which males sexually mature early are at 14L:10D, 25 ± 1 °C. These results will improve the application of S. rueppellii in crops, for the control of aphid pests.
Analyses of internal transcribed spacer 2 (ITS2) sequences (495 nucleotides) from Spissistilus festinus populations collected from various locations, years, and crops in California and the Southeastern United States. (a) Phylogeny produced by Bayesian inferential analysis (with posterior probability indicated at the node) with the crop, location, and year of collection indicated in the node names. Black dots at nodes indicate bootstrap support of at least 80%, through Maximum Likelihood analysis (1,000 bootstrap replicates). (b) Depiction of 5.8S (blue) and 28S (red) motifs denoting the boundaries of the ITS2 region (black) used as a nuclear marker for genetic variation among populations of S. festinus. (c) Segment of MUSCLE alignment of ITS2 sequences in which three nucleotide differences at positions 141, 146, and 186 were observed between S. festinus specimens from California and the Southeastern United States.
Comparative observations of Spissistilus festinus specimens collected from (top panels) soybean in Auburn, Alabama, and (bottom panels) alfalfa in Lodi, California. Full body side view (a,d), and lateral view of the male terminalia (b,e) and dissected genitalia (c,f). Note the less elevated pronotum of the specimen from California compared to the specimen from Alabama.
(a) Diagnostic polymerase chain reaction for DNA sequence-based identification of specimens of the S. festinus genotype from California (lanes 1-8) and the Southeastern United States (lanes 9-13). (b) Specificity of the diagnostic PCR against other Membracidae (lanes 1-4), Cercopidae (lanes 5-6), Fulgoroidea (lanes 7-8), Cicadellidae (lanes 9-14), Aphididae (lane 15), Aleyrodidae (lane
Sites of Spissistilus festinus collection, date of collection, mt-COI primers used for PCR characterization and sequencing, and NCBI GenBank accession numbers.
Spissistilus festinus (Say, 1830) (Hemiptera: Membracidae) is a frequent pest of leguminous crops in the Southern United States, and a vector of grapevine red blotch virus. There is currently no information on the genetic diversity of S. festinus. In this study, populations of S. festinus were collected in 2015–2017 from various crops and geographic locations in the United States, and fragments of the mitochondrial cytochrome C oxidase 1 (mt-COI) gene and the nuclear internal transcribed spacer 2 (ITS2) region were characterized by polymerase chain reaction and sequencing. Maximum-likelihood and Bayesian analyses of the mt-COI and ITS2 sequences yielded similar phylogenetic tree topologies, revealing two distinct genetic S. festinus lineages with all of the specimens from California comprising one phylogenetic clade, alongside a single GenBank entry from Arizona, and all specimens from the Southeastern United States comprising a statistically-supported distinct clade, regardless of host and year of collection. The mt-COI gene fragment showed up to 10.8% genetic distance between the two phylogenetic clades. These results suggest the existence of two genotypes within S. festinus in the United States. The only distinct morphological trait between the two genotypes was a less elevated pronotum in the representative specimens from California, compared to the representative specimens from the Southeastern United States. Since this phenotypic feature is inconspicuous, a diagnostic polymerase chain reaction targeting a variable region of the mt-COI fragment was developed to reliably distinguish between the specimens of the two genotypes of S. festinus and to facilitate their specific identification.
The cobblestone tiger beetle, Cicindelidia marginipennis (Dejean, 1831) is a North American species specializing in riparian habitats from New Brunswick, Canada, to Alabama in the United States. In the United States, this species is state-listed as threatened or endangered range-wide and periodically receives consideration for federal listing, mostly due to habitat decline. Despite its conservation status, intraspecific genetic diversity for this species has not been explored and little is known about its natural history. To support further inquiry into the biology of C. marginipennis, this study provides the first look at range-wide genetic diversity using mitochondrial DNA (mtDNA), describes all three larval instars, and describes natural history characteristics from captive rearing and field observation. Based on mtDNA analyses, our results suggest that geographically based population structure may exist throughout the range, with individuals from Alabama possessing haplotypes not found elsewhere in our sampling. Further genetic analyses, particularly multi-locus analyses, are needed to determine whether the Alabama population represents a separate cryptic species. Our morphological analysis and descriptions of larval instars reveal a combination of characteristics that can be used to differentiate C. marginipennis from closely related and co-occurring species. Based on our field observations, we find that the larval “throw pile” of soil excavated from burrows is a key search image for locating larvae, and we provide descriptions and detailed photographs to aid surveys. Lastly, we find that this species can be successfully reared in captivity and provide guidelines to aid future recovery efforts.
Top-cited authors
Almudena Ortiz-Urquiza
  • Swansea University
Nemat O Keyhani
  • University of Florida
Zareen Bharucha
  • Anglia Ruskin University
Jules N Pretty
  • University of Essex