After high population densities of insect defoliators there is often a dramatic decrease in their abundance due to various limiting factors. Here, we compared gypsy moth larvae (Lymantria dispar L.) reared singly and in crowded conditions. We compared a number of physiological parameters of these insects and the effect of L. dispar population density on the activation of covert baculovirus infections in the larvae. It was found that the population density of gypsy moth larvae did not affect the mortality due to the activation of the covert virus infection or the total mortality. On the other hand, solitary-reared larvae were heavier, took longer to develop, and showed a four-fold higher concentration of dopamine in their haemolymph than larvae reared in groups. Thus, we demonstrated that an increase in the population density of larvae per se facilitates some changes in fitness and innate immunity traits but is not related to the activation of covert baculovirus infection. We suggest that an increase in population density does not increase the risk of epizootics triggered by the activation of covert baculovirus infection and that researchers should pay more attention to studying density-associated factors, such as starvation.

The complete mitochondrial genome (mitogenome) of Spilarctia robusta (Lepidoptera: Noctuoidea: Erebidae) was sequenced and analyzed. The circular mitogenome is made up of 15,447 base pairs (bp). It contains a set of 37 genes, with the gene complement and order similar to that of other lepidopterans. The 12 protein coding genes (PCGs) have a typical mitochondrial start codon (ATN codons), whereas cytochrome c oxidase subunit 1 (cox1) gene utilizes unusually the CAG codon as documented for other lepidopteran mitogenomes. Four of the 13 PCGs have incomplete termination codons, the cox1, nad4 and nad6 with a single T, but cox2 has TA. It comprises six major intergenic spacers, with the exception of the A+T-rich region, spanning at least 10 bp in the mitogenome. The nucleotide composition of the genome is greatly A+T biased (81.09%), with a negative AT skewness (-0.007), indicating the presence of fewer As than Ts, similar to other Noctuoidea. The A+T-rich region is 343 bp long, and contains some conserved regions, including an "ATAGA" motif followed by a 19 bp poly-T stretch, a microsatellite-like (AT)₉ and a poly-A element, a characteristic shared with other lepidopteran mitogenomes. Phylogenetic analysis, based on 13 PCGs using Maximum likelihood methods revealed that S. robusta belongs to the superfamily Noctuoidea.

Insect cellular immune reaction, which generally includes phagocytosis, encapsulation and nodule formation, is achieved by hemocytes circulating in insect haemolymph. The shift of hemocytes from the normal phase to the adhering phase is an important process in the cellular immune reaction, which includes the attachment of hemocytes to foreign surfaces or other hemocytes via adhesion factors. Neuroglian is one of the adhering factors associated with encapsulation in Manduca sexta and Drosophila melanogaster. Here we studied the localization of neuroglian (MsNrg) in Mythimna separata and its functional role in the cellular immune reaction. The distribution of MsNrg mRNA between hemocyte populations was determined using real-time quantitative reverse transcription PCR and in situ hybridization, which revealed that MsNrg was highly expressed in adhering hemocytes, especially in plasmatocytes. Unexpectedly, the transcript was observed as well in non-adhering hemocytes, implying neuroglian has a function in non-adhering hemocytes. Moreover, we showed that the amount of MsNrg mRNA was not changed by injections of either biotic or abiotic non-selves. Fewer latex beads were fully encapsulated by hemocytes in larvae treated with MsNrg double-stranded RNA than in control larvae, but their ability to achieve phagocytosis and nodule formation remained unchanged by the MsNrg knockdown. These results indicate that the function of neuroglian in the cellular immune reaction is conserved in D. melanogaster and lepidopteran species, and neuroglian in non-adhering hemocytes could possess unidentified function.

Polyhydroxylated derivatives of 6-keto,7-dehydrocholesterol (ecdysteroids) are common constituents of various plants. In 1965, they were accidentally discovered in the search for the insect moulting hormone. These biologically important natural compounds are neither insect hormones nor inducers of insect ecdysis. Due to their strong anabolic, vitamin D-like effects in insects, domestic animals and humans, I propose the use of the arbitrary term vitamin D₁. The present paper describes the effects of vitamin D₁ on the growth and regeneration of excised epidermal cells of the tobacco hornworm, Manduca sexta (Sphingidae). The periods of programmed cell death and cell proliferation (histolysis and histogenesis, respectively) exactly coincide in insects with endogenous peaks of increased concentration of vitamin D₁. Epidermal cells communicate with each other, creating a mutually integrated tissue, connected by mechanical, chemical, electrical, ionic or other so far incompletely known factors. After natural cell death, or after the artificial removal of some epidermal cells, the neighbouring cells that lose communication integrity, begin to divide mitotically to replace the disconnected part. Cell divisions are arrested as soon as the integrity of the living tissue is established. During insect ontogeny, the application of juvenile hormone causes regenerating epidermal cells to repeat the previous morphogenetic programme (i.e., development of patches of larval tissue on the body of a pupa, or metathetely). Conversely, the application of vitamin D₁ (20-hydroxyecdysone) caused the regenerating cells to prematurely execute a future morphogenetic programme (i.e., development of patches of pupal tissue on the body of a larva, or prothetely). Among the key features of insect regeneration, is the arrest of cell divisions when tissues resume living cell-to-cell integrity. This prevents the formation of aberrant groups of cells, or tumours. It is well established that the main physiological systems of insects (e.g., circulatory, respiratory, neuro-endocrine) are structurally and functionally similar to corresponding systems in humans. Thus the basic principles of cell regeneration and the role of vitamin D₁ in insects may also be valid for humans. The common vitamins D₂ (ergocalciferol) or D₃ (cholecalciferol), are exclusively lipid soluble secosterols, which require activation by UV irradiation and hydroxylation in the liver. By contrast, the neglected vitamin D₁ is a natural derivative of polyhydroxylated 7-dehydrocholesterol of predominantly plant origin, which is both partly a water and partly a lipid soluble vitamin. It neither requires UV irradiation, nor hydroxylation due to 6 or 7 already built-in hydroxylic groups. Like other vitamins, it enters insect or human bodies in plant food or is produced by intestinal symbionts. Vitamin D₁ causes strong anabolic, vitamin D-like effects in domestic animals and in humans. I am convinced that avitaminosis associated with a deficiency of vitamin D₁ in human blood may be responsible for certain hitherto incurable human diseases, especially those related to impaired nerve functions and somatic growth, aberrant cell regeneration

Social wasps are often considered as nuisance pests in urban environments and are often controlled by using traps. The majority of commercially produced traps for catching wasps have yellow as the dominant colour around the trap entrance. However, the observations on the function of yellow as an attractant for wasps are controversial. The efficiency of yellow, compared with green striped (N = 15) and yellow and green striped beer traps (N = 15) was evaluated. According to the results, yellow does not have a specific role as an attractant for wasps of the genera Vespula Linnaeus and Dolichovespula (Rohwer). For wasps, it may be the bait that is the major lure and it might be sufficient on its own for both control and monitoring purposes.

The tenebrionid beetles on 25 circum-Sicilian islands were studied to determine the influence of island geographical and landscape features on three main intercorrelated biogeographical patterns: (1) species richness, studied using species-area and species environment relationships, (2) species assemblage composition, investigated using Canonical Correspondence Analysis (CCA), and (3) inter-site faunal similarity, investigated using Canonical Correlation Analysis (CANCOR) applied to multidimensional scaling of inter-island faunal dissimilarities. Species richness was mostly influenced by island area and landscape heterogeneity (expressed using various indices of diversity based on land cover categories). When species identities were considered in the CCA, no substantial effect of landscape was detected. Current island isolation did not have a strong influence on species richness, but has a distinct effect in determining species assortments on the remotest islands. Historical influences of Pleistocene landbridge connections were not detectable in species richness relationships using geographical variables in species richness analyses or in assemblage gradients in the CCA, but emerged distinctly from inter-island similarities in the CANCOR.

Larvae of the endangered Scarce Large Blue butterfly, Phengaris (Maculinea) teleius, are initially endophytic, feeding upon developing seeds of Sanguisorba officinalis, but complete their development as social parasites preying on the brood of Myrmica ants. Females show very specific preferences, laying eggs exclusively in young flower heads of the food plant. In the present contribution we report the results of a detailed study of female behaviour at three sites differing in size and abundance of S. officinalis. Tracked individuals switched between feeding on nectar (nectaring), flight and oviposition. They fed exclusively on flowers of reddish, pink or violet colouration, favouring Viccia cracca and S. officinalis. However, females spent about one fifth of the time they were active ovipositing and laid about 20 eggs per hour. Oviposition visits to flower heads were highly stereotyped and followed a specific ritual. Examination of accepted, rejected and randomly sampled flower heads showed that females avoided those containing conspecific eggs and at a site where this was difficult due to a shortage of suitable flower heads, they tended to distribute their eggs evenly among the available flower heads. Our results indirectly indicate the existence of an oviposition-deterring pheromone (ODP) in P. teleius. This hypothetical marking ensures a more even distribution of eggs and possibly increases the chances of survival of caterpillars by reducing competition during both the phytophagous and myrmecophagous stages of development. There are few reports of butterflies producing ODPs and to our knowledge this is the first evidence of their presence in the family Lycaenidae.

As urbanisation is set to continue, understanding the impact on wildlife becomes increasingly important if we are to be able to conserve biodiversity. As an excellent group of bioindicators, invertebrates can allow us to understand some of the forces in urban areas which impact upon biodiversity and wildlife populations. This paper discusses some of the trends in the abundance, diversity and richness of invertebrates related to urbanisation and the specific urban environmental and traffic factors which may be at play.

The aim of this study was to investigate the composition of the fungal community associated with the red-haired bark beetle (Hylurgus ligniperda Fabricius) in two plantations of Pinus sylvestris L. located in the Kharkiv and Luhansk regions (ca. 250 km apart) in the forest-steppe zone in eastern Ukraine. In each plantation, 48 beetles were collected from butts of living trees and 48 beetles from stems of fallen trees, i.e., a total of 96. Half of the beetles from each site were used for culturing fungi and the other half for direct sequencing the internal transcribed spacer of fungal ribosomal RNA (ITS rRNA). Thirty distinct fungal taxa were identified by culturing and 31 by direct sequencing. When pooled, there were 40 fungal taxa among which Ophiostoma piceae (Münch) Sydow & P. Sydow (10.3%), Alternaria alternata (Fries) Keissler (9.7%), Ogataea neopini Nagatsuka, S. Saito & Sugiyama (8.0%), Botryotinia fuckeliana (de Bary) Whetzel (5.1%), Cladosporium sp. Link (5.1%) and Sydowia polyspora (Brefeld & Tavel) E. Müller (4.6%) were the most common. Species of the genus Ophiostoma were the most abundant and included five different taxa O. piceae, O. bicolor R.W. Davidson & D.E. Wells, O. ips (Rumbold) Nannfeldt, O. canum (Münch) Sydow & P. Sydow and O. rectangulosporium Ohtaka, Masuya & Yamaoka, all of which are known to be at most weak pathogens of trees. The plant pathogen Botryotinia fuckeliana and insect pathogens Isaria farinose (Holmskjold) Fries and Beauveria bassiana (Balsamo-Crivelli) Vuillemin were also detected. Basidiomycetes were rare, among which three wood-decaying fungi Bjerkandera adusta (Willdenow) P. Karsten, Fomitopsis pinicola (Swartz) P. Karsten and Heterobasidion annosum (Fries) Brefeld were detected. In conclusion, in the forest-steppe zone in eastern Ukraine H. ligniperda is a vector of diverse communities of fungi the majority of which, if at all, are only weak pathogens of trees.

Knowledge of the optimal temperatures for development and survival of biological control agents is essential for efficient mass-rearing and introduction of natural enemies in augmentative biological control programs. We studied the effect of constant and alternating temperatures on development and survival of immature stages and the sex ratio at emergence of adults of the Nearctic generalist predator Geocoris punctipes (Say). We also determined its thermal requirements. They were reared in climatic chambers at alternating (21/11°C, 24/18°C, 27/21°C and 30/26°C ± 1°C) and constant temperatures (16.8°C, 21.5°C, 24.5°C and 28.3°C ± 1°C), RH 70 ± 10% and a 14 h photophase. Survival and development of G. punctipes were the same when reared at constant and alternating temperatures. Five instars were recorded in all temperature regimes. The duration of the egg stage and each instar, as well as that of total larval development were longer, and larval survival lower when reared at 16.8°C, 21/11°C, 21.5°C and 24/18°C than at 24.5°C, 27/21°C, 28.3°C and 30/26°C. The optimal temperature range for development and survival of G. punctipes is 24.5°C to 30°C, its lower development threshold temperature is 13.5°C and its thermal constant 295.9 DD. Sex ratios were not significantly different from 1 : 1 male : female ratio in all temperature regimes. There is an excellent match between the temperature regimes at which the prey Tuta absoluta (Meyrick) and predator G. punctipes are active, which indicates that this predator will function well in crops where this pest is present.

We have investigated for the first time the chromosomes of Karoophasma biedouwense, a species belonging to the Mantophasmatodea, a recently discovered order of carnivorous insects. Our study has revealed that males of this species display testes with numerous seminal tubes (follicles), as in other Polyneoptera, and short tubular seminal vesicles embedded in a utricular gland. The karyotype consists of 2n = 12A + X monocentric and biarmed, meta/submetacentric chromosomes (fundamental number of arms: FN = 26) with blocks of heterochromatin around centromeres. The autosomes are classified into two size groups, one represented by a single, very large pair of autosomes, the other by five smaller pairs which constitute a continuous series gradually decreasing in size. Among "monocentric" orders of Polyneoptera, K. biedouwense shares its low chromosome number, 2n = 13, as also found with some Orthoptera (Acridoidea, Grylloidea, Gryllacridoidea). Male meiosis is of the classical pre-reductional type and the X(0) sex determination system is probably an ancestral state. Use of FISH along with an 18S rDNA probe revealed multiple ribosomal clusters, which most likely represent an apomorphic condition. We identified the ancestral insect telomeric sequence (TTAGG)_n in the terminal areas of the chromosomes. Currently available data on the polyneopteran orders putatively related to Mantophasmatodea showed a wide variability of cytogenetic characteristics within and between them. The only character allowing some tentative inference to be made on the ancestry of K. biedouwense is its low chromosome number, the karyotypic pattern so far unreported for the Polyneoptera except in certain Orthoptera.

The adult demographic parameters, mobility, nectar choice and how the spatial distribution of males and females of Z. polyxena is affected by the distribution and abundance of host-plants, and adults of the opposite sex was studied in a population of this species inhabiting a dense network of permanent habitats (totalling 8.7 ha). The population size was estimated to be ca. 300 individuals. The average adult lifespan was 4.4 days and the maximum 23 (male) and 20 (female) days. The capture probability was higher for males than females due to the more conspicuous behaviour and bounded area of activity of males. A slow increase was followed by a slow decrease in the sex specific parabolic recruitment curve, indicating slight protandry and long emergence period, probably due to habitat heterogeneity. The spatial distribution of host plants (Aristolochia lutea) is the key factor determining the spatial distribution of adults. There was a strong positive correlation between male and female density at each patch, both of which were dependent on the cover of host plants growing in sunny conditions. In searching for A. lutea plants suitable for oviposition, females fly greater distances and move more frequently between patches than males. The size, shape and orientation of the male home range were influenced by the size, shape and orientation of stands of host-plants in sunny positions, but not by patch area. Such adult fidelity to stands of host-plants in sunny positions indicates that the spatial distributions of oviposition sites, mate-locating sites and larval habitats of Z. polyxena overlap. The better statistical fit and much lower probabilities for long-distance movements generated by a negative exponential function than an inverse power function are probably due to the small size and high habitat connectivity of the site studied. Adults were opportunistic in their use of nectar plants. Traditional management is the key factor for maintaining permanent habitats for this species in a grassland biotope.

Facultative parthenogenesis is important for mayflies with short alate stages because females are able to reproduce without mating. We studied facultative parthenogenesis in Ephoron eophilum, a mayfly with an extremely short alate stage. We examined the survival rates of embryos from unfertilized eggs, in addition to investigating the number of chromosomes in parthenogenetic offspring and the mode of inheritance by nuclear genetic analyses using Exon-Primed Intron-Crossing markers. The survival rate of thelytokous embryos was 0-70.2% (16.7 ± 26.7%, mean ± S.D.). Sixteen chromosomes were present throughout most of the mitotic metaphase in parthenogenetic offspring, which was similar to the number recorded in diploid females. All parthenogenetic offspring were homozygous in nuclear genetic analyses, despite the presence of heterozygous mothers. These results indicate that E. eophilum has the ability to reproduce via facultative parthenogenesis, producing mostly diploid thelytokous offspring. The restoration of ploidy level occurs by automixis via terminal fusion or gamete duplication, and causes rapid reduction of heterozygosity. However, despite this, significant deviation from Hardy-Weinberg equilibrium (HWE) was not observed in the studied populations. This is because facultative parthenogenesis in these circumstances normally has little influence on population genetic structuring, even though parthenogenetic embryos exhibit a high survival rate. The lack of influence of parthenogenesis on the population structure of the natural population strongly suggests that parthenogenesis rarely occurs under natural circumstances.

There are approximately 1200 described species of Chrysopidae, many of which are predators of agricultural pests. Species of Chrysoperla are mass-produced and sold for use as biological control agents of agricultural pests in Europe, Asia, North and South America. Chrysoperla agilis, a member of the "carnea group" of Chrysoperla, has the potential to be biocontrol agent and is native to the Azores, and therefore a local candidate for use in IPM of pests, such as aphids and scale insects. Given that many adult Chrysopidae feed on pollen and honeydew and the biology of Ch. agilis is not well understood, we studied the preferences of Ch. agilis adults living in a greenhouse on the campus of the University of the Azores for feeding on different types of pollen. Twenty-six species of flowering plants, belonging to 16 families and 24 genera, all previously recorded on Sao Miguel Island, were found in the greenhouse. Pollen grains from each species were collected and incorporated in a reference catalogue. Additionally, 40 adults of Ch. agilis were collected (20 in December 2012 and 20 in January 2013) and the pollen in guts identified. Of the 26 species of plants in the greenhouse 14 were used as sources of pollen by the lacewings. Pollen of Plantago lanceolata L. was the most frequently recorded pollen in the gut contents. The adults of this chrysopid fed mainly on the pollen of species of plants belonging to the Asteraceae, Plantaginaceae and Poaceae. The species richness (S = 12), diversity index (H' = 2.123) and equitability (J' = 0.8543) of the January sample are greater than those of the December sample. Similarity of the samples collected in the 2 months was 0.75. From these results we infer that it is important to maintain or increase plant biodiversity in agroecosystems as weeds provide trophic resources for insects that are beneficial to agriculture.

Northern bumblebees (Hymenoptera: Apidae) with annual lifecycles depend both on energy stores remaining in their fat body after diapause and a few spring flowering plants. Most temperate bumblebees emerge gradually over several months after winter depending on the species and within species on the location of the overwintering chamber (hibernaculum). Weather can either delay or promote emergence and nectar resources are needed to fuel flight at low ambient temperatures to find a nest site. Several phoretic mites use queens for transportation and have synchronized life cycles with their host species. Their presence on the body of bumblebees is usually harmless but can become harmful when the number of mites increases to hundreds per individual bee. High numbers of mites on queens may also indicate the queens are in poor condition. The effect of temperature and mite load on the time for which queens can survive (d) during a 25-day period of starvation were determined using newly emerged spring queens of B. lucorum. Queens collected from various locations in SW Finland were divided into four groups taking into account their initial load of Parasitellus fucorum mites: (A) 15°C and provided with only water; (B) 24°C and provided with only water; (C) 15°C and provided with pollen and nectar; (D) 24°C and provided with pollen and nectar. There were mites on 65% (50 out of 77) of the queens. All of the fed queens survived and starved queens died, with those kept at 24°C dying approximately 8 days before those kept at 15°C. Queen weight or mite load had no effect on the length of the period for which they survived and there was no difference in the weights of the queens in the different mite load classes. Asynchrony between plants and insect as well as increased frost damage due to climate change may affect nectar availability in spring. Therefore, the survival and long term viability of bumblebee populations should be monitored during variable spring conditions.

Carabid beetles could potentially provide a number of ecosystem services in arable fields, including the regulation of weeds by eating seeds. Spatio-temporal variability in the incidence of predation combined with a limited understanding of the interactions between carabids and seed predation, currently limits our ability to assess the effectiveness of this service. We carried out a long-term field study of carabid communities and the incidence of predation of the seeds of 5 species of weeds and laboratory cafeteria choice-tests to quantify the preferences of the most abundant species of seed-eating carabid for the seeds of 10 species of weeds. The field study revealed important temporal variation both in carabid activity-density and seed predation, with the seed of Viola arvensis and Capsella bursa-pastoris being particularly preferred by the carabids. The laboratory cafeteria tests confirmed the predation preferences measured in the field, in showing strong preferences by both Pterostichus melanarius and Pseudoophonus rufipes for the seed of V. arvensis. The percentage predation of the seed of V. arvensis was associated with the activity-density of carabids, while that of the seed of C. bursa-pastoris was not. Integrating laboratory-measured daily consumption rates, per carabid taxa, into models improved the ability to predict the intensity of predation of the seed of V. arvensis based on the composition of carabid communities.

Knowledge of the dispersal ability of endangered species is crucial for developing effective, evidence-based conservation policies. Due to their limited dispersal abilities and specific habitat requirements, insects are among the animals most threatened by habitat fragmentation. We studied three populations of the highly endangered species of ground beetle, Carabus hungaricus, at three sites in Central Europe (Hungary and Czech Republic) using mark-release-recapture (MRR). The total catch of 574 pitfall traps set at the three sites was 6255 individuals. Depending on the site, the percentage recaptured was 13-32%. Average and maximum distance moved by individuals of both sexes at each of the sites ranged between 47-132 and 207-1104 m, respectively. The probability of the movements following an inverse power function (IPF) for the two sexes did not differ, but did differ among sites. Probability of dispersing for distances >100 m differed by an order of magnitude between sites, most likely because of differences in how the samples were collected. Despite the fact that individual beetles are able to move over distances in the order of kilometres, the high fragmentation of their habitats is likely to prevent them from colonizing most uninhabited habitat patches. Therefore, the conservation of this threatened ground beetle could be improved by adopting and implementing a policy of assisted dispersal. Our results from three study sites also provide an interesting illustration of the variability in the estimates of the probability of dispersal obtained using MRR.

Laboratory experiments were conducted to examine the effects of photoperiod and temperature on the pupation behaviour of the parasitoid, Microplitis mediator (Haliday) (Hymenoptera: Braconidae), parasitizing larvae of Mythimna separata Walker (Lepidoptera: Noctuidae). A combination of long photoperiod (14 + h L) and warm temperatures (20-24°C) caused parasitized caterpillars to climb to upper plant parts where the parasitoid produced a green, non-diapausing cocoon on a green leaf, initially retaining the dying caterpillar host as a protective covering. In contrast, short photoperiod (8-10 h L) and low temperature (16-18°C) induced host caterpillars to descend the plant where the parasitoid produced a brown, diapausing cocoon either hanging by silk from a senescing leaf or simply lying on the soil, but without any continued association with the host. These findings illustrate the potential for seasonal environmental cues to simultaneously mediate diapause induction, cocoon polymorphism, and alternate forms of host behaviour modification in a hymenopterous parasitoid.

Parasitic Hymenoptera usually need to have direct access to their host or its offspring or to be able to enter and place their offspring in a nest of their host, where their progeny develop at the expense of the host. In the case of the cleptoparasitic cuckoo wasps (Chrysididae) their main target is the nest of the host and they have various strategies for locating and entering their host's brood cells for ovipositing. The chrysidid Omalus biaccinctus (Buysson, 1893), a parasitoid of aphid hunting crabronid wasps, was observed apparently inserting its eggs into living aphids at the hunting site of their host and the presence of the parasitoid's egg and DNA in aphids after oviposition was confirmed. It is concluded that O. biaccinctus "hitch-hikes" into a host's brood cell concealed in the aphid prey collected by the host wasp and the adult parasites do not need to enter the nest of their host in order to lay their eggs.

In order to understand patterns of abundances of migratory Lepidoptera in southernmost Europe and contrast this with those of sedentary species, we studied butterflies surveyed along transects during three spring migration seasons at the Rock of Gibraltar. Overall, 2508 butterflies belonging to 19 species were recorded. Of these, the four most numerous species accounted for almost 88% of all individuals recorded. These were the migratory Clouded Yellow Colias croceus, Red Admiral Vanessa atalanta and Painted Lady Vanessa cardui, and the sedentary Common Blue Polyommatus icarus. There was a significant correlation between abundance of P. icarus and C. croceus. Furthermore, abundances of C. croceus and V. atalanta were correlated. The sometimes very high abundance of migratory butterflies at the study site could suggest that Gibraltar is a stopover site for butterflies migrating between Africa and Europe.

Prey preference of natural enemies is an important parameter used in studies on their efficiency. Feeding preferences of individuals from two populations of Propylea quatuordecimpunctata (L.) (Coleoptera: Coccinellidae) for the essential prey items, Aphis fabae Scopoli, 1763 and Aphis gossypii Glover, 1877 (Homoptera: Aphididae) were evaluated in the laboratory using Manly's β preference index (β). For the predator preference experiment, equal numbers of A. fabae and A. gossypii were offered to the predator. The experiment was conducted on a broad bean leaf disc with 20 replicates. After 24 h, the unconsumed aphids were counted. Individuals of P. quatuordecimpunctata from both populations consumed more A. gossypii [Karaj population (mean ± SE): 28.15 ± 1.04 and Sari population 34.35 ± 0.51] than A. fabae (Karaj population: 17.95 ± 0.64 and Sari population: 17.7 ± 1.01). The values of Manly's β preference index (β) for A. gossypii were 0.73 ± 0.01 and 0.77 ± 0.01 for the Karaj and Sari populations, respectively and for A. fabae were 0.26 ± 0.01 and 0.22 ± 0.01 for the Karaj and Sari populations, respectively. Comparison of the preference indices using a t-test revealed significant differences between the prey preferences of those from the Sari population (t = 22.53, df = 38, P < 0.0001) and the Karaj population (t = 26, df = 38, P < 0.0001). These results reveal that P. quatuordecimpunctata from the two populations prefer A. gossypii over A. fabae.

The ribosomal protein S6 kinase (S6K) plays a pivotal role in developmental processes and cell survival by participating in protein synthesis relevant signaling pathways. In the present study, an S6K gene (AccS6K-p70) was isolated and characterized from the Chinese honeybee, Apis cerana cerana (Fabricius) (Hymenoptera: Apidae), an important economic insect in the agricultural industry. The cDNA of AccS6K-p70 was 1683 bp in length and predicted to encode a protein of 467 amino acid residues. Sequence and structure analysis showed that there was a conserved catalytic domain in AccS6K-p70, whilst a phosphorylation site was found in the conserved part of the catalytic domain. Development relevant transcription factor binding sites found in the 5'-flanking region of AccS6K-p70 suggest that AccS6K-p70 might be involved in A. c. cerana development. Furthermore, quantitative PCR revealed that the expression levels of AccS6K-p70 were higher in head and thorax than in other tissues. The AccS6K-p70 was highly expressed in both larvae and adults compared with that in pupae, whilst expression of the gene was significantly down-regulated by hydrogen peroxide (H₂O₂) (although initially and slightly increased by it) and pyriproxyfen (a juvenile hormone analogue insecticide) stresses. These results suggest that AccS6K-p70 may play critical roles in developmental processes and cell survival in A. c. cerana, whilst both oxidative stress and pyriproxyfen may impair S6K-p70 mediated developmental processes by down-regulation of AccS6K-p70 expression.

A novel panel of 16 microsatellite markers, obtained by pyrosequencing of enriched genomic libraries, is reported for the flightless European bushcricket Ephippiger diurnus (Dufour) (Orthoptera: Tettigoniidae). Five multiplex and one simplex PCR protocols were optimized, and the polymorphism at the 16 loci was assessed in two natural populations from southern France. The mean allele number and (expected mean heterozygosity) were 8.94 (0.71) and 6.57 (0.70), respectively, in each population. Several loci were at Hardy-Weinberg disequilibrium (HWD), possibly due to the incidence of null alleles. The occurrence of null alleles has been previously reported for this species, and it is a common feature of microsatellite loci in Orthoptera. Cross-amplification tests demonstrated the transferability of some of these loci to other ephippigerine species. The microsatellite loci reported here substantially increase the number of available loci for this species and will afford an accurate picture of E. diurnus phylogeography, the genetic structure of its populations, and an improved understanding of the evolution of male song and other sexually-selected traits in this highly variable species.

Coordinated regulation of apoptosis is critical for development, homeostasis, and immunity in larvae of Metazoa. We determined the full nucleotide sequence of an inhibitor of an apoptosis protein in a lepidopteran insect Mythimna separata (Walker) (MsIAP) and carried out functional analyses of the MsIAP. The full-length cDNA of MsIAP was 1642 bp, which encoded 379 amino acid residues with a calculated molecular mass of 41,834 Da, and two BIR domains and one RING domain revealed using amino acid sequence analysis. In addition, the sequences of these domains were similar to Drosophila IAP1 and those of some other lepidopteran insects. We carried out a functional analyses of MsIAP related to apoptosis regulation using RNA interference. The effects of MsIAP knockdown on adhering hemocytes and non-adhering hemocytes as controls were examined using Hoechst33342/propidium iodide staining, effector caspase activity and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) staining. A significantly higher number of propidium iodide and TUNEL-positive cells was recorded in adhering hemocytes from MsIAP knockdown larvae than from control larvae, but these differences were not recorded for non-adhering hemocytes. However, higher effector caspase activity was detected in both adhering and non-adhering hemocytes from MsIAP knockdown larvae compared to that in control larvae. These results indicate that the knockdown of MsIAP induces apoptosis in larval adhering hemocytes, which MsIAP negatively and non-redundantly regulate apoptosis, and that IAP function is conserved in M. separata and other insect species including Drosophila and several lepidopteran insects.

Brazilian native meliponines are currently threatened by increased human impacts. The assessment of their genetic variation by microsatellite DNA markers can assist in the conservation of populations and help in the planning and establishment of efficient management strategies. The purpose of this study was to develop the first set of microsatellite markers for Melipona fasciculata, selected from partial genome assembly of Illumina paired-end reads. Primer pairs were designed for each detected locus at their flanking regions. Bee samples were genotyped from two different populations of Northeastern Brazil for marker characterization and validation. A total of 17 microsatellite loci displayed polymorphism. Mean H_E and H_O heterozygosities were 0.453 and 0.536, respectively. PIC across all loci ranged from 0.108 to 0.714. A genetic diversity analysis revealed high values for population differentiation estimates (F_ST = 0.194, R_ST = 0.230, and D_est = 0.162) within the investigated region. PCoA and Bayesian clustering showed a separation of the species into two distinct clusters. These microsatellite markers have demonstrated strong potential for population-level genetic studies. Moreover, the preliminary analysis of the genetic diversity in M. fasciculata provides provisional evidence of significant population differentiation between the two studied populations.

Members of the clade Trichophora (Hemiptera: Heteroptera: Pentatomomorpha) have trichobothria on their abdominal sterna. There is no comparative study of the fine structure of abdominal trichobothria in the group and until now the trichobothria of their immatures were virtually unknown. The fine structure of the abdominal trichobothrial complex (= the trichobothrium and its associated structures) of adults of 98 species belonging to 25 families in 5 superfamilies and larvae of 7 species belonging to 7 families in 2 superfamilies of Trichophora were examined using scanning electron microscopy. This study indicates that the fine structure of the abdominal trichobothria is very variable and useful for determining evolutionary lineages within the clade. Six types of bothria, three of trichomes and three of microtrichia are recognized and their evolutionary transformations discussed. Changes in the size of trichomes, and density and size of the microtrichia during the postembryonic development of selected species are discussed.

The leafhopper genus Alebra Fieber, 1872 comprises a complex of morphologically similar species. The chromosome complements (karyotypes) of five Alebra species, i.e. A. albostriella, A. coryli, A. viridis, A. wahlbergi and a new, yet undescribed species, provisionally named Taxon 1, were here investigated, three of these species (A. coryli, A. viridis, and Taxon 1) for the first time. The techniques applied included standard chromosome staining, fluorescence in situ hybridization (FISH) for mapping of 18S rDNA and telomeric repeats (in every species), C-banding, AgNOR-banding and CMA₃/DAPI- staining (in A. viridis). The species have a holokinetic type of chromosomes, as in other hemipterans. Karyotypes of all species are remarkably conserved with 2n = 22 + X(0)/XX (male/female), one large and 10 medium pairs of autosomes and the X chromosome similar in size to larger chromosomes within this group. In every species, FISH identified the "classical" insect telomere repeat of TTAGG and rRNA gene clusters located on the homologues of a medium-sized pair of autosomes, presumably number 5. Thus, speciation in Alebra has apparently not involved significant karyotypic changes. In A. viridis, rDNA sites were both Ag- and CMA₃-positive and were located at an interstitial position. C-banding revealed heterochromatic bands in the X chromosome and also in all but four pairs of autosomes, the bands were located at one telomere of a chromosome. C-bands were positive for CMA₃ and negative for DAPI, suggesting that C-heterochromatin is mainly enriched in GC-pairs.

The objective of this work was to evaluate the relative trap efficiency of the three modified traps baited with 1-octen-3-ol as attractant, Box, Nzi and Canopy, for horse fly (Tabanidae) survey. Nine traps (three traps per trap type) were tested in 3 × 3 Latin square designs during summer 2011 in riparian oak-ash forests in the Danube floodplain of Croatia. Overall, the traps caught 1,986 specimens of 11 horse flies species during the study period. The most abundant species caught was Tabanus bromius (58%), followed by Tabanus tergestinus (21%), Tabanus maculicornis (16%), Tabanus sudeticus (2%), Atylotus loewianus (1%) and Tabanus autumnalis (1%). Both the Box traps and the Nzi traps had significantly greater catch efficiencies than the Canopy traps (β_box = 2.47, X² = 65.48, df = 1, P < 0.001 and β_nzi = 1.54, X² = 25.12, df = 1, P < 0.001, respectively). Nzi traps had a catch rate 4.65 (95% CI: 2.55, 8.48) times greater than Canopy traps and Box traps had a catch rate 11.77 (95% CI: 6.48, 21.39) times greater than Canopy traps. Based on Shannon-Wiener diversity indices Nzi traps were better suited for diversity survey and had higher (H' = 1.33) indices than Box (H' = 1.08) or Canopy traps (H' = 1.00).

We studied the water balance, body fluid osmolality and survival of the oribatid mite, Pergalumna nervosa, when exposed to drought in field and laboratory experiments. In a replicated field experiment we artificially lowered the soil water content by putting roofs over selected plots, which reduced soil water potential to levels well below the permanent wilting percentage for plants (i.e. below -1.5 MPa). Even though a slight decrease in the abundance of P. nervosa (only found in the 0-5 cm soil layer) was recorded during the most severe drought stress (ca. -3.5 MPa), the majority of adult mites clearly survived these conditions for 3 weeks in the field without migrating to deeper soil layers. Exposing field collected adults in laboratory experiments simulating even more severe drought conditions revealed that P. nervosa can survive several weeks of gradually increasing drought stress (down to -7 MPa) with moderate water loss. The osmolality of body fluids increased as dehydration progressed, but apparently as a result of simple up-concentration of solutes and not the de novo synthesis of protective osmolytes. We compare and discuss these results in the light of what is known about other arthropods.

The expression of sexually dimorphic phenotypes from a shared genome between males and females is a longstanding puzzle in evolutionary biology. Increasingly, research has made use of transcriptomic technology to examine the molecular basis of sexual dimorphism through gene expression studies, but even this level of detail misses the metabolic processes that ultimately link gene expression with the whole organism phenotype. We use metabolic profiling in Drosophila melanogaster to complete this missing step, with a view to examining variation in male and female metabolic profiles, or metabolomes, throughout development. We show that the metabolome varies considerably throughout larval, pupal and adult stages. We also find significant sexual dimorphism in the metabolome, although only in pupae and adults, and the extent of dimorphism increases throughout development. We compare this to transcriptomic data from the same population and find that the general pattern of increasing sex differences throughout development is mirrored in RNA expression. We discuss our results in terms of the usefulness of metabolic profiling in linking genotype and phenotype to more fully understand the basis of sexually dimorphic phenotypes.