The European (Melolontha melolontha L.) and Forest (M. hippocastani F.) cockchafer are widespread pests throughout Central Europe. Both species exhibit a 3–5-year life cycle and occur in temporally shifted populations, which have been monitored and documented for more than 100 years. Visual identification of adults and larvae belonging to these morphologically similar species requires expertise and, particularly in the case of larvae, is challenging and equivocal. The goal of the study was the development of an efficient and fast molecular genetic tool for the identification and discrimination of M. melolontha and M. hippocastani. We established a collection of both species from Switzerland, Austria and Northern Italy in 2016, 2017 and 2018. An approximately 1550 bp long fragment of the cytochrome c oxidase subunit 1 (CO1) mitochondrial gene was amplified and sequenced in 13 M. melolontha and 13 M. hippocastani beetles. Alignment of the new sequences with reference sequences (NCBI GenBank and BOLDSYSTEMS databases) and subsequent phylogenetic analysis revealed consistent clustering of the two species. After the identification of M. melolontha and M. hippocastani species-specific single nucleotide polymorphisms (SNPs) in the CO1 alignment, we developed an effective SNP tool based on the ABI PRISM® SNaPshot™ Multiplex Kit for the rapid and accurate species discrimination of adults and larvae.

Dryocosmus kuriphilus (Hymenoptera: Cynipidae) is a global invasive gall wasp and a pest of chestnuts (Castanea spp.). A study of the Chalcidoidea parasitoid community of D. kuriphilus was undertaken over two years, from March 2017 to March 2019, at 15 sites in south and northwest Spain (Málaga and Galicia regions). More than 18,000 galls were collected, and 1153 parasitoids belonging to 22 species of seven chalcidoid families, plus two individuals of an inquiline Cynipidae, Synergus facialis, emerged. Richness was higher in the Málaga region, with 20 species, while 17 parasitoids and one inquiline were identified in Galicia. The parasitism rate of native chalcid parasitoid species in both regions was low. Eupelmus urozonus and Mesopolobus lichtensteini were the most abundant native species. Mesopolobus tibialis was a dominant species in south Spain, while Ormyrus pomaceus was a dominant species in northwest Spain. Our results revealed the existence of a sub-community of univoltine, probably host specialized, parasitoids in south Spain, which overwinter in galls, exhibiting a similar life cycle to Torymus sinensis. These species were Torymus notatus, Aulogymnus bicolor, Aulogymnus obscuripes and Aulogymnus balani. Data on the recovery of T. sinensis after release in the south Spain region show it to be well established, but its numbers are still low in northwest Spain.

The brown marmorated stink bug Halyomorpha halys is one of the most harmful invasive species in the world. Native to East Asia, this insect was introduced into North America in the 1990s and into Europe in the 2000s where it subsequently established and spread across the continent. Previous population genetic studies determined the invasion pathways at continental and national levels. However, information on the dynamics on a small-scale is currently scarce. Here we study the genetic diversity and population dynamics of H. halys in South Tyrol, a region in Northern Italy, since its arrival to its widespread establishment over a period of four years. By haplotyping 162 individuals from ten populations (including six previously published individuals) we found a high haplotype diversity in most populations with an increasing diversity across the different years. Most haplotypes were previously found in other regions of Northern Italy, providing evidence for migration from neighboring regions. However, the presence of four previously undescribed haplotypes as well as a haplotype previously found exclusively in Greece highlights additional long-distance dispersal across the continent. Phylogenetic analysis of the haplotypes found in South Tyrol showed that the majority of haplotypes clustered with haplotypes predominantly found in Japan. This suggests a potential recent introduction of H. halys individuals from Japan into Europe, and thus an additional invasion pathway that was previously unidentified.

For a wide range of insect species, the microbiota has potential roles in determining host developmental programme, immunity and reproductive biology. The tea geometrid moths Ectropis obliqua and E. grisescens are two closely related species that mainly feed on tea leaves. Although they can mate, infertile hybrids are produced. Therefore, these species provide a pair of model species for studying the molecular mechanisms of microbiotal involvement in host reproductive biology. In this study, we first identified and compared the compositions of microbiota between these sibling species, revealing higher microbiotal diversity for E. grisescens. The microbiota of E. obliqua mainly comprised the phyla Firmicutes, Proteobacteria and Cyanobacteria, whereas that of E. grisescens was dominated by Proteobacteria, Actinobacteria and Firmicutes. At the genus level, the dominant microbiota of E. grisescens included Wolbachia, Enterobacter and Pseudomonas and that of E. obliqua included Melissococcus, Staphylococcus and Enterobacter. Furthermore, we verified the rate of Wolbachia to infect 80 samples from eight different geographical populations, and the results supported that only E. grisescens harboured Wolbachia. Taken together, our findings indicate significantly different microbiotal compositions for E. obliqua and E. grisescens, with Wolbachia possibly being a curial factor influencing the reproductive isolation of these species. This study provides new insight into the mechanisms by which endosymbiotic bacteria, particularly Wolbachia, interact with sibling species.

Ovipositional decisions in herbivorous insects may be affected by social information from conspecifics. Social facilitation of oviposition has been suggested for the onion fly Delia antiqua. In the current study, we found that D. antiqua oviposition was unequal between paired oviposition stations of equal quality and that more eggs were laid on an oviposition station baited with decoy flies than on the control. The increased oviposition toward the decoys continued over time >8 h. When decoys were placed upside down, the number of eggs laid did not differ between the decoy and control sides of oviposition stations, suggesting that social facilitation of oviposition is mediated by visual cues. Based on these findings, mechanisms of social facilitation of oviposition in D. antiqua were discussed.

The Bemisia tabaci species complex is one of the most important pests of open field and protected cropping globally. Within this complex, one species (Middle East Asia Minor 1, B. tabaci MEAM1, formerly biotype B) has been especially problematic, invading widely and spreading a large variety of plant pathogens, and developing broad spectrum pesticide resistance. Here, we fit a CLIMEX model to the distribution records of B. tabaci MEAM1, using experimental observations to calibrate its temperature responses. In fitting the model, we consider the effects of irrigation and glasshouses in extending its potential range. The validated niche model estimates its potential distribution as being considerably broader than its present known distribution, especially in the Americas, Africa and Asia. The potential distribution of the fitted model encompasses the known distribution of B. tabaci sensu lato, highlighting the magnitude of the threat posed globally by this invasive pest species complex and the viruses it vectors to open field and protected agriculture.

The European cherry fruit fly, Rhagoletis cerasi (Diptera: Tephritidae), is a univoltine species that undergoes obligatory summer–winter diapause at pupal stage in the soil (2–5 cm) beneath host trees. To study the effects of photoperiod and relative humidity on diapause termination and post-winter developmental duration of R. cerasi, pupae collected from Dossenheim (Germany) were exposed to different photoperiod or relative humidity regimes during a chilling period ranging from 2 to 8.5 months. Specifically, pupae were exposed to four photoperiod regimes: (a) light conditions (24L:00D), (b) dark conditions (00L:24D), (c) short photoperiod (08L:16D) and (d) long photoperiod (16L:08D), as well as to three relative humidity regimes: (a) low (40% RH), (b) medium (60% RH) and (c) high (70–80% RH). Data revealed that relative humidity is not a significant predictor of diapause termination, but it affects the post-winter developmental period. Higher relative humidity promotes post-winter pupae development. On the other hand, photoperiod significantly affected both diapause termination and post-winter development of R. cerasi pupae. Light conditions (24L:00D) accelerate adult emergence, particularly for females. Regardless of the photoperiod (24L:00D, 00L:24D, 08L:16D), rates of adult emergence were high (>75%) for chilling intervals longer than 6.5 months. Nonetheless, exposure to a long day photoperiod (16L:08D), during chilling, dramatically reduced the proportion of adult emergence following 6 months exposure to chilling. Our findings broaden the understanding of factors regulating diapause responses in European cherry fruit fly, local adaptation and synchronization of adult emergence with the ripening period of major hosts.

Tribolium castaneum is one of the most economically important insects that damages stored products. The effects of several infested or uninfested raw feed materials (wheat bran, coarse wheat meal, corn feed flour), feed products (compound feed for pigs and for laying hens) and flour mixed with brewer's yeast on the food-searching behaviour of T. castaneum adults were studied in a total of 48 combinations. Preference and olfactometer tests revealed that all the tested uninfested and intraspecific infested substrates were significantly more attractive to T. castaneum than the control (represented by part of an arena or olfactometer arm without substrate). We determined that all infested substrates were 2–9 times more attractive than uninfested in the preference test, while in the olfactometer test, they were 3–8 times more attractive. In comparing the attractiveness of the infested and uninfested substrates, in both tests wheat bran was found to be the most attractive substrate to T. castaneum adults and coarse wheat meal the least. The results of the present study contribute to our knowledge of how raw feed materials and products influence the behaviour of T. castaneum and their susceptibility to infestation, and indicate the possible utilization of wheat bran in monitoring processes of T. castaneum in pest management programmes.

Tuta absoluta (Meyrick), a key pest of tomato, is quickly spreading over the world and biological control is considered as one of the control options. Worldwide more than 160 species of natural enemies are associated with this pest, and an important challenge is to quickly find an effective biocontrol agent from this pool of candidate species. Evaluation criteria for control agents are presented, with the advantages they offer for separating potentially useful natural enemies from less promising ones. Next, an aggregate parameter for ranking agents is proposed: the pest kill rate km. We explain why the predator's intrinsic rate of increase cannot be used for comparing the control potential of predators or parasitoids, while km can be used to compare both types of natural enemies. As an example, kill rates for males, females and both sexes combined of three Neotropical mirid species (Campyloneuropsis infumatus (Carvalho), Engytatus varians (Distant) and Macrolophus basicornis (Stål)) were determined, taking all life-history data (developmental times, survival rates, total nymphal and adult predation, sex ratios and adult lifespan) into account. Based on the value for the intrinsic rate of increase (rm) for T. absoluta and for the kill rate km of the predators, we predict that all three predators are potentially able to control the pest, because their km values are all higher than the rm of the pest. Using only km values, we conclude that E. varians is the best candidate for control of T. absoluta on tomato, with C. infumatus ranking second and M. basicornis last.

The white-backed planthopper, Sogatella furcifera (Horváth) (Hemiptera, Delphacidae), has emerged as a serious rice pest in Asia. In the present study, 12 microsatellite markers were employed to investigate the genetic structure, diversity and migration route of 43 populations sampled from seven Asian countries (Bangladesh, China, Korea, Laos, Nepal, Thailand, and Vietnam). According to the isolation by distance analysis, a significant positive correlation was observed between genetic and geographic distances by the Mantel test (r2 = 0.4585, P = 0.01), indicating the role of geographic isolation in the genetic structure of S. furcifera. A population assignment test using the first-generation migrants detection method (thresholds a = 0.01) revealed southern China and northern Vietnam as the main sources of S. furcifera in Korea. Nepal and Bangladesh might be additional potential sources via interconnection with Vietnam populations. This paper provides useful data for the migration route and origin of S. furcifera in Korea and will contribute to planthopper resistance management.

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a pest species complex that causes widespread damage to cassava, a staple food crop for millions of households in East Africa. Species in the complex cause direct feeding damage to cassava and are the vectors of multiple plant viruses. Whilst significant work has gone into developing virus-resistant cassava cultivars, there has been little research effort aimed at understanding the ecology of these insect vectors. Here we assess critically the knowledge base relating to factors that may lead to high population densities of sub-Saharan African (SSA) B. tabaci species in cassava production landscapes of East Africa. We focus first on empirical studies that have examined biotic or abiotic factors that may lead to high populations. We then identify knowledge gaps that need to be filled to deliver sustainable management solutions. We found that whilst many hypotheses have been put forward to explain the increases in abundance witnessed since the early 1990s, there are little published data and these tend to have been collected in a piecemeal manner. The most critical knowledge gaps identified were: (i) understanding how cassava cultivars and alternative host plants impact population dynamics and natural enemies; (ii) the impact of natural enemies in terms of reducing the frequency of outbreaks and (iii) the use and management of insecticides to delay the development of resistance. In addition, there are several fundamental methodologies that need to be developed and deployed in East Africa to address some of the more challenging knowledge gaps.

Trialeurodes vaporariorum (Westwood, 1856) (Greenhouse whitefly) is an agricultural pest of global importance. It is associated with damage to plants during feeding and subsequent virus transmission. Yet, global phylogenetic relationships, population structure, and estimation of the rates of gene flow within this whitefly species remain largely unexplored. In this study, we obtained and filtered 227 GenBank records of mitochondrial cytochrome c oxidase I (mtCOI) sequences of T. vaporariorum, across various global locations to obtain a final set of 217 GenBank records. We further amplified and sequenced a ~750 bp fragment of mtCOI from an additional 31 samples collected from Kenya in 2014. Based on a total of 248 mtCOI sequences, we identified 16 haplotypes, with extensive overlap across all countries. Population structure analysis did not suggest population differentiation. Phylogenetic analysis indicated the 2014 Kenyan collection of samples clustered with a single sequence from the Netherlands to form a well-supported clade (denoted clade 1a) nested within the total set of sequences (denoted clade 1). Pairwise distances between sequences show greater sequence divergence between clades than within clades. In addition, analysis using migrate-n gave evidence for recent gene flow between the two groups. Overall, we find that T. vaporariorum forms a single large group, with evidence of further diversification consisting primarily of Kenyan sequences and one sequence from the Netherlands forming a well-supported clade.

The endoparasitoid Microplitis prodeniae Rao and Chandry is an important potential augmentative biological control agent for lepidopteran pests of vegetables and tobacco. However, cold storage of pupae is required to ensure that sufficient parasitoids are available when they are needed in the field. In this study, pupae were maintained at 0, 4 or 10°C for 5–50 days after which the adults were evaluated for emergence, pre-emergence period, sex ratio, female longevity, oviposition period, and fecundity. Cold storage did not affect the pre-emergence period or proportion of females; however, there was a significant reduction in emergence, female longevity, oviposition period, and fecundity with increased exposure to cold. The pre-emergence period was approximately 5 days, and approximately 50% of the emergent parasitoids were females. A cold storage regime of 10 days at 10°C had no effect on the parasitoids and adult emergence was greater than 50% even after 20 days at 10°C. There was no carryover of the cold treatment from parental to F1 and F2 generations. Thus, M. prodeniae can be stockpiled for field release by exposing the pupae to a cold regime and subsequently holding them for adult emergence at 28°C.

Since 1980, mosquito breeding habitats in the Upper Rhine Valley were routinely treated with Bacillus thuringiensis var. israelensis (Bti). Bti is considered to significantly reduce the number of mosquitoes, and – especially when used in higher dosages – to be toxic to other Nematocera species, e.g. Chironomidae, which could be food sources for aerial feeding predators. To investigate direct and indirect effects of routine Bti treatment on food sources for aerial feeding predators, the availability of flying insects in treated and untreated areas was compared. A car trap was used for insect collection, which allowed their exact spatiotemporal assignment. The statistical analysis revealed that insect taxa abundance was influenced differently by the factors season, temperature and time of day. Nematocera (Diptera) were the most frequently collected insects in all areas. Chironomidae were the predominant aquatic Nematocera. The comparison of treated and untreated sites did not show significant differences that would indicate any direct or indirect effect of routine Bti treatment on the availability of flying insects. Additional to food availability, food selection must be considered when investigating food resources for aerial feeding predators. In this study, food selection of Delichon urbicum (House Martin) as an example was investigated with the help of neck ring samples. The preferred prey of the investigated D. urbicum colony consisted of diurnal insects with terrestrial larvae (Aphidina, Brachycera, Coleoptera). Chironomidae were consumed, but not preferred.

Ectropis grisescens Warren and Ectropis obliqua (Prout) are two morphologically similar sibling species with overlapping ranges. In this study, manipulative laboratory experiments were conducted to examine the possibility of reproductive interference in sympatric populations of E. grisescens and E. obliqua and the potential consequences of the mating interaction. Our results showed that the presence of males or females of different species could incur mating interference and significant reduction of F1 offspring. The reduction was not significant relevant to the initial relative abundance of E. grisescens and E. obliqua. Detailed observations of mating opportunity showed that female mating frequencies of both species were not significantly affected by the absolute species density, but the mating success of E. obliqua females with conspecific males depended on species ratio. In addition, adding males to the other species resulted in lower number of offspring suggesting that the males’ behaviour might be linked with mating interference. Males of both E. grisescens and E. obliqua could interfere the intraspecific mating of the other species, but the impact of the mating interference differed. These combined data indicated that asymmetric reproductive interference existed in E. grisescens and E. obliqua under laboratory conditions, and the offspring of the mixed species were significantly reduced. The long term outcome of this effect is yet to be determined since additional reproductive factors such as oviposition rate and progeny survival to adulthood may reduce the probability of demographic displacement of one species by the other in overlapping niches.

In Australia Bt cotton has been planted since 1996, and has greatly improved the control of its key target Helicoverpa armigera (Hübner). There is no strong evidence that genetically modified cotton has been selected for significant physiological resistance to Bt toxin in field populations. There are many possible explanations for the lack of apparent selection that range from high compliance with the resistance management strategy for this technology to a lack of behavioral preference in key traits such as oviposition that could favor survival. To date most experiments that test oviposition of H. armigera on Bt cotton vs. conventional cotton have been done with susceptible moths. We determine the oviposition preference of a field isolated Bt resistant line of H. armigera and a susceptible counterpart when given a choice of non-Bt cotton and Bt-cotton with the same genetic background, and test whether there is any relationship between oviposition site selection (different plant structures) and the survival of the first instar larvae. Within cotton plants, our experiments consistently showed that both resistant and susceptible moths did not choose plants or plant parts that were less toxic in terms of Bt toxin on which to lay eggs. There was one exception in that susceptible moths were more likely to lay eggs on squares of Bt cotton plants than squares of non-Bt cotton. As expected, the mortality of susceptible H. armigera neonates was significantly higher on structures of Bt cotton plants than on those structures of conventional cotton, and survival was greater on flowers than on other structures of Bt cotton. This confirms opportunities for selection for resistance, and demonstrates no advantage in this respect to carrying resistance genes that might overcome the Bt toxins.

Callosobruchus chinensis (Linnaeus) is one of the most destructive pests of leguminous seeds. Genetic differentiation and diversity analysis of 345 C. chinensis individuals from 23 geographic populations using 20 polymorphic simple sequence repeats revealed a total of 149 alleles with an average of 7.45 alleles per locus. The average Shannon's information index was 1.015. The gene flow and genetic differentiation rate values at the 20 loci ranged from 0.201 to 1.841 and 11.0–47.2%, with averages of 0.849 and 24.4%, respectively. In the 23 geographic populations, the effective number of alleles and observed heterozygosity ranged from 1.441 to 2.218 and 0.191–0.410, respectively. Shannon's information index ranged from 0.357 to 0.949, with the highest value in Hohhot and the lowest in Rudong. In all comparisons, the fixation index (FST) values ranged from 0.049 to 0.441 with a total FST value of 0.254 among the 23 C. chinensis populations, indicating a moderate level of genetic differentiation and gene flow among these populations. Analysis of molecular variance revealed that the genetic variation within populations accounted for 76.7% of the total genetic variation. The genetic similarity values between populations varied from 0.617 to 0.969, whereas genetic distances varied from 0.032 to 0.483. Using unweighted pair-group method using arithmetical averages cluster analysis, the 23 geographic collections were classified into four distinct genetic groups but most of them were clustered into a single group. The pattern of the three concentrated groups from polymerase chain reactions analysis showed a somewhat different result with cluster.

The distribution of phlebotomine sand flies is widely reported to be changing in Europe. This can be attributed to either the discovery of sand flies in areas where they were previously overlooked (generally following an outbreak of leishmaniasis or other sand fly-related disease) or to true expansion of their range as a result of climatic or environmental changes. Routine surveillance for phlebotomines in Europe is localized, and often one of the challenges for entomologists working in non-leishmaniasis endemic countries is the lack of knowledge on how to conduct, plan and execute sampling for phlebotomines, or how to adapt on-going sampling strategies for other haematophagous diptera. This review brings together published and unpublished expert knowledge on sampling strategies for European phlebotomines of public health concern in order to provide practical advice on: how to conduct surveys; the collection and interpretation of field data; suitable techniques for the preservation of specimens obtained by different sampling methods; molecular techniques used for species identification; and the pathogens associated with sand flies and their detection methods.

Brown marmorated stink bug, Halyomorpha halys (Stål), is a highly polyphagous invasive species attacking both cultivated and wild plants increasing its threat to ecosystems as a global pest. However, dispersal biology of this invasive species is not well understood. This study evaluated the flight capacity and behavior of H. halys under laboratory, semi-field, and field conditions. Flight mills were used to measure the baseline flight capacity of adults collected year round from the field and included both foraging and overwintering populations. The effects of abiotic conditions such as wind speed and temperatures on the free flight parameters of H. halys were evaluated under semi-field and field conditions. The mean flight distances over a 22-h period were 2442 and 2083 m for male and female, respectively. Most individuals (89%) flew <5 km, though some flew much further with a maximum flight distance observed of 117 km. Flight distances by H. halys increased after emergence from overwintering sites in spring and reached their highest point in June. The incidence of take off by H. halys was significantly affected by the wind speed; when provided with still air conditions, 83% of individuals took off, but the rates decreased to <10% when wind speed was increased to or above 0.75 m s−1. The incidence of take off by H. halys was significantly affected by ambient temperature and light intensity in the field, whereas relative humidity and insect sex did not. When the temperature was at 10–15°C, 3% of individuals took off, but the proportion of H. halys taking flight increased to 61, 84, and 87% at 15–20, 20–25, and 25–30°C, respectively. In the field, prevailing flight direction was biased toward the opposite direction of the sun's position, especially in the morning. The implications of H. halys flight biology are discussed in the context of developing monitoring and management programs for this invasive species.

The Australian plague locust, Chortoicetes terminifera (Walker), is an important agricultural pest and oviposits into compacted soil across vast semi-arid and arid regions prone to irregular heavy summer rainfall. This study aimed to quantify the effects of flooding (control, 7, 14, 21, 28 and 35 days) at different temperatures (15, 20 and 25°C) and embryonic development stages (25 and 75%) on egg viability, hatchling nymph body mass and survival to second-instar. Egg viability after flooding was dependent on temperature and flood duration. Eggs inundated at 15°C showed ≥53.5% survival regardless of flood duration and development stage compared with ≤29.6% for eggs at 25°C for ≥21 days early in development and ≥14 days late in development. Hatchling nymphs did not differ in body mass relative to temperature or flood duration, but weighed more from eggs inundated early in development rather than late. Survival to second-instar was ≤55.1% at 15 and 20°C when eggs were flooded for ≥28 days late in development, ≤35.6% at 25°C when flooded for ≥28 days early in development, and zero when flooded for ≥21 days late in development. These results suggest that prolonged flooding in summer and early autumn may cause very high egg mortality and first-instar nymph mortality of any survivors, but is likely to only ever affect a small proportion of the metapopulation. More common flash flooding for ≤14 days is unlikely to cause high mortality and have any direct effect on distribution and abundance.