Our work aims to assess how butterfly communities in the Italian Maritime Alps changed over the past 40 years, in parallel with altitudinal shifts occurring in plant communities. In 2019, we sampled butterflies at 7 grassland sites, between 1300–1900 m, previously investigated in 2009 and 1978, by semi-quantitative linear transects. Fine-scale temperature and precipitation data elaborated by optimal interpolation techniques were used to quantify climate changes. The changes in the vegetation cover and main habitat alterations were assessed by inspection of aerial photographs (1978–2018/1978–2006–2015). The vegetation structure showed a marked decrease of grassland habitats and an increase of woods (1978–2009). Plant physiognomy has remained stable in recent years (2009–2019) with some local exceptions due to geomorphic disturbance. We observed butterfly ‘species substitution’ indicating a general loss in the more specialised and a general gain in more tolerant elements. We did not observe any decrease in species richness, but rather a change in guild compositions, with (i) an overall increased abundance in some widespread and common lowland species and (ii) the disappearance (or strong decrease) of some alpine (high elevation) species, so that ‘resilience’ could be just delusive. Changes in butterfly community composition were consistent with predicted impacts of local warming. Full article

Many arachnid taxa have evolved unique, highly specialized sensory structures such as antenniform legs in Amblypygi (whip spiders), for instance, or mesosomal pectines in scorpions. Knowledge of the neuroanatomy as well as functional aspects of these sensory organs is rather scarce, especially in comparison to other arthropod clades. In pseudoscorpions, no special sensory structures have been discovered so far. Nevertheless, these animals possess dominant, multifunctional pedipalps, which are good candidates for being the primary sensory appendages. However, only little is known about the anatomy of the nervous system and the projection pattern of pedipalpal afferents in this taxon. By using immunofluorescent labeling of neuronal structures as well as lipophilic dye labeling of pedipalpal pathways, we identified the arcuate body, as well as a comparatively small mushroom body, the latter showing some similarities to that of Solifugae (sun spiders and camel spiders). Furthermore, afferents from the pedipalps terminate in a glomerular and a layered neuropil. Due to the innervation pattern and structural appearance, we conclude that these neuropils are the first integration centers of the chemosensory and mechanosensory afferents. Within Arthropoda, but also other invertebrates or even vertebrates, sensory structures show rather similar neuronal arrangement. Thus, these similarities in the sensory systems of different evolutionary origin have to be interpreted as functional prerequisites of the respective modality. Full article

Agriculture is dependent on insect pollination, yet in areas of intensive production agriculture, there is often a decline in plant and insect diversity. As native habitats and plants are replaced, often only the weeds or unwanted vegetation persist. This study compared insect diversity on mango, Mangifera indica, a tropical fruit tree dependent on insect pollination, when weeds were present in cultivation versus when they were removed mechanically. The pollinating insects on both weeds and mango trees were examined as well as fruit set and yield in both the weed-free and weedy treatment in South Florida. There were significantly more pollinators and key pollinator families on the weedy mango trees, as well as significantly greater fruit yield in the weedy treatment compared to the weed-free treatment. Utilizing weeds, especially native species, as insectary plants can help ensure sufficient pollination of mango and increase biodiversity across crop monocropping systems. Full article

High morphological variation is often associated with species longevity, and it is hypothesized that urban-dwelling species may require more plasticity in functional traits such as body size in order to maximize fitness in heterogeneous environments. There has been published research regarding the functional trait diversity of urban bee pollinators. However, no two cities are identical, so the implementation of multi-city studies is vital. Therefore, we compared body size variation in female Halicus ligatus sweat bees from May–October 2016 from three distinct Midwestern United States cities: Chicago, Detroit, and Saint Louis. Additionally, to elucidate potentially influential environmental factors, we assessed the relationship between temperature and measured body size. We collected bees in community gardens and urban farms and measured their head width and intertegular distance as a proxy for overall body size. We utilized an ANCOVA to determine whether body size variation differed significantly across the three surveyed cities. Results indicated that H. ligatus females in Chicago, Detroit, and Saint Louis had significantly different body size ranges. These findings highlight the importance of intraspecific body size variation and support our prediction that bees from different urban environments will have distinct ranges in body size due to local ecological factors affecting their populations. Additionally, we found a significant influence of temperature, though this is probably not the only important ecological characteristic impacting bee body size. Therefore, we also provided a list of predictions for the future study of specific variables that are likely to impact functional trait diversity in urban bees. Full article

Ant-like flies comprise nine Iberian endemic species of flightless Tachydromia. Severe knowledge gaps on distribution and ecological requirements hinder conservation assessments. Species distribution models were applied to unveil habitat suitability and to provide guidelines for future studies. An ensemble modeling approach combining ten different techniques was implemented with the biomod2 package. Occurrence data was partitioned into six sets, including two multi-species groups and four species. The most relevant drivers of habitat suitability are climate-related, followed by forest type and structure, according to well-defined biogeographic gradients. T. lusitanica and T. ebejeri are adapted to mild temperatures and high-humidity environments. Their distribution is connected to the Temperate–Eurosiberian life zone. T. semiaptera and T. iberica are adapted to progressively drier and hotter central and southern parts of the Iberian Peninsula, connected to transitional Temperate–submediterranean areas. Ant-like fly’ ranges overlap with deciduous/marcescent oak species, acting as suitable indicators of their presence in Iberia. Southern marcescent forests emerge as “islands” with particular interest for future prospections. Ant-like flies are threatened by several factors such as climate change and habitat destruction, including urbanization and forest fires. This study provides vital tools to better assess the ant-like flies’ conservation status and to manage their habitat. Full article

Condition-specific competition, when environmental conditions alter the outcome of competition, can foster the persistence of resident species after the invasion of a competitively superior invader. We test whether condition-specific competition can facilitate the areawide persistence of the resident and principal West Nile virus vector mosquito Culex pipiens with the competitively superior invasive Aedes albopictus in water from different urban container habitats. (2) Methods: We tested the effects of manipulated numbers of A. albopictus on C. pipiens’ survival and development in water collected from common functional and discarded containers in Baltimore, MD, USA. The experiment was conducted with typical numbers of larvae found in field surveys of C. pipiens and A. albopictus and container water quality. (3) Results: We found increased densities of A. albopictus negatively affected the survivorship and development of C. pipiens in water from discarded containers but had little effect in water from functional containers. This finding was driven by water from trash cans, which allowed consistently higher C. pipiens’ survival and development and had greater mean ammonia and nitrate concentrations that can promote microbial food than other container types. (4) Conclusions: These results suggest that the contents of different urban containers alter the effects of invasive A. albopictus competition on resident C. pipiens, that trash cans, in particular, facilitate the persistence of C. pipiens, and that there could be implications for West Nile virus risk as a result. Full article

Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is an important economic pest of citrus crops because it vectors the causal pathogen of huanglongbing (HLB; aka citrus greening). Population suppression of D. citri with insecticides has been disproportionally relied on for HLB management and a greater diversity of more sustainable tools is needed. Periglandula spp. is a fungal endosymbiont (family Clavicipitaceae) that forms a mutualistic relationship with members of plants in family Convolvulaceae. This association results in the production of ergot alkaloids that were previously documented as having psyllicidal properties. We investigated the mortality and behavior of D. citri exposed to crude extracts from morning glories in the plant family Convolvulaceae, as well as synthetic ergot alkaloids. Nymphs and adults were exposed to the crude plant extracts from Periglandula positive species of Convolvulaceae, as well as five synthetic ergot alkaloids. Treatments were prepared by exposing clippings of citrus to 100 ng/µL of crude extract from Periglandula-positive species of Ipomoea (I. imperati, I. leptophylla, I. pandurata and I. tricolor), and Turbina corymbosa, and from one Periglandula-negative species (I. alba) (100 ng/µL). Mortality of adult and nymphal D. citri was significantly higher than the control after exposure to extracts from I. tricolor and I. imperati. The synthetic ergot alkaloids, lysergol (10–100 ng/µL), ergonovine maleate (100 ng/µL), agroclavine (10–100 ng/µL), and ergosine (10–100 ng/µL) increased mortality of D. citri nymphs, while ergosine (100 ng/µL) and agroclavine (100 ng/µL) increased mortality of adults compared to water controls. Fewer D. citri adults settled on plants treated with crude extracts or synthetic ergot alkaloids than on water controls at 48 h after release. D. citri that fed on citrus leaves treated with 10 ng/μL solution of crude extract from the Periglandula-positive species Ipomoea (I. imperati, I. leptophylla, I. pandurata, I. tricolor), and Turbina corymbosa excreted significantly less honeydew compared with a negative water control and extract from Periglandula-negative species (I. alba). Our results indicate that crude extracts and ergot alkaloids exhibit toxic and sub-lethal effects on D. citri that could be useful for management of this pest. Full article

The adaptation by planthoppers to feed and develop on resistant rice is a challenge for pest management in Asia. We conducted a series of manipulative experiments with the brown planthopper (Nilaparvata lugens (Stål)) on the resistant rice variety IR62 (BPH3/BPH32 genes) to assess behavioral and bionomic changes in planthoppers exhibiting virulence adaptation. We also examined the potential role of yeast-like symbionts (YLS) in virulence adaptation by assessing progeny fitness (survival × reproduction) following controlled matings between virulent males or females and avirulent males or females, and by manipulating YLS densities in progeny through heat treatment. We found virulence-adapted planthoppers developed faster, grew larger, had adults that survived for longer, had female-biased progeny, and produced more eggs than non-selected planthoppers on the resistant variety. However, feeding capacity—as revealed through honeydew composition—remained inefficient on IR62, even after 20+ generations of exposure to the resistant host. Virulence was derived from both the male and female parents; however, females contributed more than males to progeny virulence. We found that YLS are essential for normal planthopper development and densities are highest in virulent nymphs feeding on the resistant host; however, we found only weak evidence that YLS densities contributed more to virulence. Virulence against IR62 in the brown planthopper, therefore, involves a complex of traits that encompass a series of behavioral, physiological, and genetic mechanisms, some of which are determined only by the female parent. Full article

On tropical mountains, predation pressure decreases with elevation. Accordingly, one expects an elevational decay in the prevalence of costly defensive traits such as aposematic coloration. Using light-trap catches of Arctiinae moths (353 species, 4466 individuals), assembled along a forested gradient in the megadiverse tropical Andes of southern Ecuador, we show that the incidence of aposematic coloration decreases strongly between 1040 and 2670 m asl. While over 60% of Arctiinae moths were warningly colored at lowest sites, this fraction decreased to less than 20% in montane forest, yet increased slightly again at the highest sites in the very open Purdiaea nutans forest. In parallel, the incidence of hymenopteran mimics and of species that mimic chemically defended beetles decreased with elevation. Hymenopteran mimics accounted for less than 5% of Arctiinae moths at sites above 2100 m, and beetle mimics were essentially lacking at high elevations. These patterns coincide with a change in gross taxonomic composition of Arctiinae ensembles and with an increase in average body size towards higher elevations. Representatives of Euchromiina and Ctenuchina became scarce with altitude, whereas the prevalence of Lithosiinae increased. Our findings suggest that the variable selective pressures along the elevational gradient favor warning coloration primarily at lower sites, whereas cryptic appearance of adult moths dominates in the tropical upper montane forest. Full article

Aedes aegypti is the principal mosquito vector for many arthropod-borne viruses (arboviruses) including dengue, chikungunya, and Zika. In the United States, excessive permethrin use has led to a high frequency of resistance in mosquitoes. Insecticide resistance is a significant obstacle in the struggle against vector-borne diseases. To help overcome metabolic resistance, synergists that inhibit specific metabolic enzymes can be added to formulated pyrethroid products. Using modified CDC bottle bioassays, we assessed the effect of three inhibitors (piperonyl butoxide (PBO), which inhibits oxidase activity; S.S.S-tributyl phosphorotrithioate (DEF), which inhibits esterase activity; and diethyl maleate (DM), which inhibits glutathione transferase activity) + permethrin. We performed these against 20 Florida Ae. aegypti populations, all of which were resistant to permethrin. Our data indicated that 11 out of 20 populations (55%) exhibited metabolic resistance. Results revealed 73% of these populations had significant increases in mortality attributed to DEF + permethrin, 64% to PBO + permethrin, and 55% to DM + permethrin compared to permethrin alone. Currently, PBO is the only metabolic enzyme inhibitor added to formulated pyrethroid products used for adult mosquito control. Our results suggest that the DEF and DM inhibitors could also be useful additives in permethrin products, especially against metabolically resistant Ae. aegypti mosquitoes. Moreover, metabolic assays should be conducted to better inform mosquito control programs for designing and implementing integrated vector management strategies. Full article

Developmental temperature plays important roles in the expression of insect traits through thermal developmental plasticity. We exposed the aphid parasitoid Aphidius colemani to different temperature regimes (10, 20, or 28 °C) throughout larval development and studied the expression of morphological and physiological traits indicator of fitness and heat tolerance in the adult. We showed that the mass decreased and the surface to volume ratio of parasitoids increased with the development temperature. Water content was not affected by rearing temperature, but parasitoids accumulated more lipids when reared at 20 °C. Egg content was not affected by developmental temperature, but adult survival was better for parasitoids reared at 20 °C. Finally, parasitoids developed at 20 °C showed the highest heat stupor threshold, whereas parasitoids developed at 28 °C showed the highest heat coma threshold (better heat tolerance CTmax1 and CTmax2, respectively), therefore only partly supporting the beneficial acclimation hypothesis. From a fundamental point of view, our study highlights the role of thermal plasticity (adaptive or not) on the expression of different life history traits in insects and the possible correlations that exist between these traits. From an applied perspective, these results are important in the context of biological control through mass release techniques of parasitoids in hot environments. Full article

Insecticide resistance is an informative model for studying the appearance of adaptive traits. Simultaneously, understanding how many times resistance mutations originate is essential to design effective resistance management. In the mosquito Culex pipiens, target–site resistance to the insecticide diflubenzuron (DFB) has been recently found in Italian and Turkish populations. Three point mutations confer it at the codon 1043 of the chitin synthase 1 gene (chs-1): I1043L, I1043M, and I1043F. Whether the resistant mutations originated independently from different susceptible alleles or sequentially from resistant alleles and whether resistant alleles from Italy and Turkey have originated once or multiple times remain unresolved. Here, we sequenced a fragment of the chs-1 gene carrying the resistant mutations and inferred the phylogenetic relationships among susceptible and resistant alleles. Confirming previous findings, we found the three mutations in Italy and the I1043M in Turkey. Notably, the I1043F was also found for the first time in Turkish samples, highlighting the need for extensive monitoring activities. Phylogenetic analyses are consistent with an independent origin of the I1043F, I1043M, and I1043L mutations from different susceptible alleles and with multiple independent origins of the Italian and Turkish I1043M and I1043F alleles. Full article

Honey bee (Apis mellifera) health is impacted by viral infections at the colony, individual bee, and cellular levels. To investigate honey bee antiviral defense mechanisms at the cellular level we further developed the use of cultured primary cells, derived from either larvae or pupae, and demonstrated that these cells could be infected with a panel of viruses, including common honey bee infecting viruses (i.e., sacbrood virus (SBV) and deformed wing virus (DWV)) and an insect model virus, Flock House virus (FHV). Virus abundances were quantified over the course of infection. The production of infectious virions in cultured honey bee pupal cells was demonstrated by determining that naïve cells became infected after the transfer of deformed wing virus or Flock House virus from infected cell cultures. Initial characterization of the honey bee antiviral immune responses at the cellular level indicated that there were virus-specific responses, which included increased expression of bee antiviral protein-1 (GenBank: MF116383) in SBV-infected pupal cells and increased expression of argonaute-2 and dicer-like in FHV-infected hemocytes and pupal cells. Additional studies are required to further elucidate virus-specific honey bee antiviral defense mechanisms. The continued use of cultured primary honey bee cells for studies that involve multiple viruses will address this knowledge gap. Full article

Garments treated with chemical insecticides are commonly used to prevent mosquito bites. Resistance to insecticides, however, is threatening the efficacy of this technology, and people are increasingly concerned about the potential health impacts of wearing insecticide-treated clothing. Here, we report a mathematical model for fabric barriers that resist bites from Aedes aegypti mosquitoes based on textile physical structure and no insecticides. The model was derived from mosquito morphometrics and analysis of mosquito biting behavior. Woven filter fabrics, precision polypropylene plates, and knitted fabrics were used for model validation. Then, based on the model predictions, prototype knitted textiles and garments were developed that prevented mosquito biting, and comfort testing showed the garments to possess superior thermophysiological properties. Our fabrics provided a three-times greater bite resistance than the insecticide-treated cloth. Our predictive model can be used to develop additional textiles in the future for garments that are highly bite resistant to mosquitoes. Full article

The entomophilous pollination niche (abundance, phenotypic traits, foraging behaviours and environmental tolerances of insect pollinators) helps to understand and better manage crop pollination. We apply this niche approach to assess how an entomophilous crop (blueberry, Vaccinium ashei) can be expanded into new territories (i.e., northern Spain) far from their original area of domestication (North America). Insect visits to blueberry flowers were monitored in a plantation on 12 different days, at 8 different times during day and covering various weather conditions. Abundance, visitation rate, pollen gathering behaviour, and frequency of inter-plant and inter-row movements were recorded. The pollinator assemblage was basically composed of one managed honeybee species (50.8% of visits) and three native bumblebee species (48.3%). There was a marked pattern of seasonal segregation throughout bloom, with bumblebees dominating the early bloom and honeybee the late bloom. Pollinators also segregated along gradients of daily temperature and relative humidity. Finally, the two pollinator types differed in foraging behaviour, with bumblebees having a visitation rate double that of honeybee, collecting pollen more frequently and changing plant and row more frequently. The spatio-temporal and functional complementarity between honeybee and bumblebees suggested here encourages the consideration of an integrated crop pollination strategy for blueberries, based on the concurrence of both wild and managed bees. Full article

Arundo donax (giant reed) is invasive in Mediterranean, sub-, and tropical riparian systems worldwide. The armored scale Rhizaspidiotus donacis is approved for biocontrol in North America, but an adventive population was recently discovered in southern California. We documented this population’s distribution, phylogeny, phenology, potential host spillover to Phragmites spp., and potential for parasitism by a common biocontrol parasitoid of citrus scale. The adventive scale was found within a single watershed and is genetically closest to Iberian scale genotypes. Rhizaspidiotus donacis developed on Phragmites haplotypes but at much lower densities than Arundo. The adventive population is univoltine, producing crawlers from March-June. Aphytis melinus parasitoids exhibited sustained interest in R. donacis during choice and no-choice trials and oviposition resulted in a small second generation. Rhizaspidiotus donacis appears limited in distribution by its univoltinism and sessile adult females. This presents challenges for broad biocontrol implementation but allows for targeted application. The genetic differentiation between imported biocontrol samples and adventive populations presents an opportunity for exploring benefits of hybrids and/or alternative genotypes where establishment has been difficult. While unlikely to occur in situ, spillover to vulnerable endemic Phragmites or deleterious parasitoid effects on scale biocontrol agents warrants consideration when planning use of R. donacis. Full article

Most organisms are dependent on sensory cues from their environment for survival and reproduction. Fireflies (Coleoptera: Lampyridae) represent an ideal system for studying sensory niche adaptation due to many species relying on bioluminescent communication; as well as a diversity of ecologies. Here; using transcriptomics; we examine the phototransduction pathway in this non-model organism; and provide some of the first evidence for positive selection in the phototransduction pathway beyond opsins in beetles. Evidence for gene duplications within Lampyridae are found in inactivation no afterpotential C and inactivation no afterpotential D. We also find strong support for positive selection in arrestin-2; inactivation no afterpotential D; and transient receptor potential-like; with weak support for positive selection in guanine nucleotide-binding protein G(q) subunit alpha and neither inactivation nor afterpotential C. Taken with other recent work in flies; butterflies; and moths; this represents an exciting new avenue of study as we seek to further understand diversification and constraint on the phototransduction pathway in light of organism ecology. Full article

Wireworms are yellowish soil-dwelling larvae that damage a wide range of arable crops. The most common wireworms found in European cultivated fields (except for the Caucasus) belong to the genus Agriotes (Coleoptera: Elateridae). In several European countries, environment-impacting insecticides are applied on a prophylactic basis to control them. However, before any treatment can be applied, European legislation requires that an assessment is done when pest population levels exceed a damage threshold. The threshold substantially depends on wireworm species, thus quick reliable larval identification is needed to implement the appropriate integrated pest management practices. Furthermore, research into non-chemical strategies involves carrying out tests with live and identified wireworms. Thus, thousands of wireworms were observed in a bid to identify live larvae so that larval density could be assessed and compared with species-specific thresholds before sowing, and laboratory experiments were carried out. This work led to a horizontal identification table that allows for quick and accurate identification of live larvae. This key, unlike traditional dichotomous keys, simultaneously considers a set of multiple discriminating morphological characters in order of stability. The key can be reliably used by less experienced users and, once minimum familiarity is acquired, most larvae can be identified rapidly, with high precision. Full article

The Culex pipiens (Diptera: Culicidae) mosquito is of high medical importance as it is considered the prime vector of West Nile virus. In Europe, this species consists of two forms, named pipiens and molestus, that exhibit substantial differences in their overwintering biology. Diflubenzuron (DFB) and Bacillus thuringiensis var. israelensis (Bti) are two of the most used larvicides in mosquito control, including that of Culex pipiens. The high dependency on these two larvicides poses major concerns for resistance development. The evolution and stability of resistance to insecticides has been associated with fitness costs that may be manifested under stressful conditions, such as the winter period. This study investigated how short-term selection of pipiens and molestus forms to both larvicides affect their overwintering success. Larvae from each form were subjected to the same selective pressure (80% mortality) for three successive generations with DFB and Bti. At the end of this process, the winter survival between the selected populations and the controls (colonies without selection) was determined for each form. Selection to both larvicides significantly reduced the winter survival rates of molestus but not of pipiens form, indicating potential differences in the persistence of the selected individuals from year to year between the two forms. Full article

Specialisation and plasticity are important for many forms of collective behaviour, but the interplay between these factors is little understood. In insect societies, workers are often developmentally primed to specialise in different tasks, sometimes with morphological or physiological adaptations, facilitating a division of labour. Workers may also plastically switch between tasks or vary their effort. The degree to which developmentally primed specialisation limits plasticity is not clear and has not been systematically tested in ecologically relevant contexts. We addressed this question in 20 free-foraging bumble bee (Bombus terrestris) colonies by continually manipulating colonies to contain either a typically diverse, or a reduced (“homogeneous”), worker body size distribution while keeping the same mean body size, over two trials. Pooling both trials, diverse colonies produced a larger comb mass, an index of colony performance. The link between body size and task was further corroborated by the finding that foragers were larger than nurses even in homogeneous colonies with a very narrow body size range. However, the overall effect of size diversity stemmed mostly from one trial. In the other trial, homogeneous and diverse colonies showed comparable performance. By comparing behavioural profiles based on several thousand observations of individuals, we found evidence that workers in homogeneous colonies in this trial rescued colony performance by plastically increasing behavioural specialisation and/or individual effort, compared to same-sized individuals in diverse colonies. Our results are consistent with a benefit to colonies of large and small specialists under certain conditions, but also suggest that plasticity or effort can compensate for reduced (size-related) specialisation. Thus, we suggest that an intricate interplay between specialisation and plasticity is functionally adaptive in bumble bee colonies. Full article

Understanding the role of facultative endosymbionts on the host’s ecology has been the main aim of the research in symbiont–host systems. However, current research on host–endosymbiont dynamics has failed to examine the genetic background of the hosts and its effect on host–endosymbiont associations in real populations. We have addressed the seasonal dynamic of facultative endosymbiont infections among different host clones of the grain aphid Sitobion avenae, on two cereal crops (wheat and oat) and whether their presence affects the total hymenopteran parasitism of aphid hosts at the field level. We present evidence of rapid seasonal shifts in the endosymbiont frequency, suggesting a positive selection of endosymbionts at the host-level (aphids) through an agricultural growing season, by two mechanisms; (1) an increase of aphid infections with endosymbionts over time, and (2) the seasonal replacement of host clones within natural populations by increasing the prevalence of aphid clones closely associated to endosymbionts. Our results highlight how genotypic variation of hosts can affect the endosymbiont prevalence in the field, being an important factor for understanding the magnitude and direction of the adaptive and/or maladaptive responses of hosts to the environment. Full article

Habitat simplification and intensive use of pesticides are main drivers of global arthropod declines and are, thus, decreasing natural pest control. Organic farming, complex landscapes, and local vineyard management practices such as implementation of flower-rich cover-crop mixtures may be a promising approach to enhance predator abundance and, therefore, natural pest control. We examined the effect of organic versus integrated management, cover-crop diversity in the vineyard inter-rows, and landscape composition on the natural pest control of Lobesia botrana eggs and pupae. Predation of L. botrana pupae was reduced by organic farming and species-poor cover-crops by about 10%. Predation rates of L. botrana eggs did not differ significantly in any of the studied management options. Dominant predators were earwigs (Forficulidae), bush crickets (Tettigoniidae), and ants (Formicidae). Negative effects of organic viticulture are most likely related to the negative nontarget effects on arthropods related to the frequent sulfur and copper applications in combination with the avoidance of strongly damaging insecticides by integrated winegrowers. While a 10% difference in predation rates on a single pest stage is unlikely to have strong practical implications, our results show that the assumed effectiveness of environmentally friendly agriculture needs to be evaluated for specific crops and regions. Full article

Aedes aegypti control requires dedicated resources that are usually scarce, limiting the reach and sustainability of vector control programs. This generates a need to focus on areas at risk of disease transmission and also understand the factors that might modulate local mosquito abundance. We evaluated the eco-bio-social factors that modulate indoor and outdoor relative abundance of female Ae. aegypti in communities of South Texas. We conducted housing quality and Knowledge Attitudes and Practices surveys in households that were part of a weekly mosquito surveillance program in November of 2017 and 2018. Our results showed widespread knowledge of mosquitoes and Zika virus by our participants. However, less than 35% considered them as serious problems in this region. The presence of window-mounted air conditioning units increased the risk of female mosquito relative abundance indoors. An increase in outdoor relative abundance was associated with larger properties and a higher number of children between 6 to 17 years of age. Interestingly, we observed that an increasing number of children <5 years of age modulated both indoor and outdoor relative abundance, with a 52% increase indoors and 30% decrease outdoors. The low perception of mosquito and disease risk highlights engagement needs for vector-borne disease prevention in this region. The identified risk factors can help guide public health officials in their efforts to reduce human and vector contact. Full article

The fall migration of monarch butterflies, Danaus plexippus (L.), in the Pacific Northwest was studied during 2017–2019 by tagging 14,040 captive-reared and 450 wild monarchs. One hundred and twenty-two captive-reared monarchs (0.87%) were recovered at distances averaging 899.9 ± 98.6 km for Washington-released and 630.5 ± 19.9 km for Oregon-released monarchs. The greatest straight-line release to recovery distance was 1392.1 km. A mean travel rate of 20.7 ± 2.2 km/day and maximum travel of 46.1 km/day were recorded. Recovery rates were greater for Oregon-released monarchs (0.92%) than Washington-released (0.34%) or Idaho-released monarchs (0.30%). Most monarchs (106/122) were recovered SSW-S-SSE in California, with 82 at 18 coastal overwintering sites. Two migrants from Oregon were recovered just weeks after release ovipositing in Santa Barbara and Palo Alto, CA. Two migrants released in central Washington recovered up to 360.0 km to the SE, and recoveries from Idaho releases to the S and SE suggests that some Pacific Northwest migrants fly to an alternative overwintering destination. Monarchs released in southern Oregon into smoky, poor quality air appeared to be as successful at reaching overwintering sites and apparently lived just as long as monarchs released into non-smoky, good quality air. Migration and lifespan for monarchs infected with the protozoan parasite, Ophryocystis elektroscirrha (McLaughlin and Myers), appeared to be similar to the migration and survival of uninfected monarchs, although data are limited. Our data improve our understanding of western monarch migration, serving as a basis for further studies and providing information for conservation planning. Full article

Co-evolution is a major driving force shaping the outcome of host-parasite interactions over time. After host shifts, the lack of co-evolution can have a drastic impact on novel host populations. Nevertheless, it is known that Western honey bee (Apismellifera) populations can cope with host-shifted ectoparasitic mites (Varroa destructor) by means of natural selection. However, adaptive phenotypic traits of the parasites and temporal variations in host resistance behavior are poorly understood. Here, we show that mites made adaptive shifts in reproductive strategy when associated with resistant hosts and that host resistance traits can change over time. In a fully-crossed field experiment, worker brood cells of local adapted and non-adapted (control) A.mellifera host colonies were infested with mites originating from both types of host colonies. Then, mite reproduction as well as recapping of cells and removal of infested brood (i.e., Varroa Sensitive Hygiene, VSH) by host workers were investigated and compared to data from the same groups of host colonies three years earlier. The data suggest adaptive shifts in mite reproductive strategies, because mites from adapted hosts have higher probabilities of reproduction, but lower fecundity, when infesting their associated hosts than mites in treated colonies. The results confirm that adapted hosts can reduce mite reproductive success. However, neither recapping of cells nor VSH were significantly expressed, even though the latter was significantly expressed in this adapted population three years earlier. This suggests temporal variation in the expression of adaptive host traits. It also appears as if mechanisms not investigated here were responsible for the reduced mite reproduction in the adapted hosts. In conclusion, a holistic view including mite adaptations and studies of the same parasite/host populations over time appears overdue to finally understand the mechanisms enabling survival of V.destructor-infested honey bee host colonies. Full article

In eusocial insects (e.g., ants, bees, and termites), the roles of different castes are assigned to different individuals. These castes possess unique phenotypes that are specialized for specific tasks. The acquisition of sterile individuals with specific roles is considered a requirement for social evolution. In termites, the soldier is a sterile caste. In primitive taxa (family Archotermopsidae and Stolotermitidae), however, secondary reproductives (neotenic reproductives) with their mandibles developed into weapons (so-called reproductive soldiers, also termed as soldier-headed reproductives or soldier neotenics) have been reported. To understand the developmental mechanism of this unique caste, it is necessary to understand the environmental cues and developmental processes of reproductive soldiers under natural conditions. Here, we established efficient conditions to induce reproductive soldiers in Zootermopsis nevadensis. Male reproductive soldiers frequently developed after the removal of both the king and soldiers from an incipient colony. Similarly, high differentiation rates of male reproductive soldiers were observed after king-and-soldier separation treatment using wire mesh. However, no male reproductive soldiers were produced without direct interaction with the queen. These results suggest that male reproductive soldier development is repressed by direct physical interactions with both the king and soldiers and facilitated by direct physical interaction with the queen. Full article

Arthropod-borne viruses (arboviruses) infect mosquito salivary glands and then escape to saliva prior to virus transmission. Arbovirus transmission from mosquitoes can be modulated by salivary gland infection barriers (SGIBs) and salivary gland escape barriers (SGEBs). We determined the influence of SGIBs and SGEBs by estimating the quantitative genetic contributions of Aedes aegypti half-sib families (Mapastepec, Mexico) infected with three dengue 2 (DENV2), two chikungunya (CHIKV), and two Zika (ZIKV) genotypes. We determined virus titer per salivary gland and saliva at seven days post-infection and virus prevalence in the half-sib population. CHIKV or ZIKV genotypes did not present SGIB, whereas DENV2 genotypes showed low rates of SGIB. However, virus titer and prevalence due to additive genetic factors in the half-sib family displayed a significant narrow-sense heritability (h²) for SGIB in two of the three DENV2 genotypes and one CHIKV and one ZIKV genotype. SGEBs were detected in all seven virus strains: 60–88% of DENV2 and 48–62% of CHIKV or ZIKV genotype infections. SGEB h² was significant for all CHIKV or ZIKV genotypes but not for any of the DENV2 genotypes. SGIBs and SGEBs exhibited classical gene-by-gene interaction dynamics and are influenced by genetic factors in the mosquito and the virus. Full article

The status of the Tramea species present in the Galapagos Islands (Odonata, Libellulidae) has been the subject of a long-standing debate among odonatologists. Here, we use molecular and morphological data to analyze a series of specimens from this genus collected in 2018 from the Islands of San Cristobal, Isabela, and Santa Cruz, with the aim of determining their relationship with Tramea calverti Muttkowski and with their currently considered senior synonym T. cophysa Hagen. We combined sequencing of mitochondrial and nuclear DNA with morphological examination of several specimens of Tramea, including representatives of continental T. cophysa and T. calverti. Our molecular analyses place the Tramea from Galapagos in the same clade as T. calverti, with T. cophysa as a closely related species. The morphological analyses found only one consistent difference between T. cophysa and T. calverti: the presence of an accessory lobe in the male vesica spermalis of T. cophysa that is absent in T. calverti and in the Tramea from Galapagos. In agreement with our genetic results, the overall morphological differences documented by us indicate that the Galapagos material examined is conspecific with T. calverti. In light of this, and following the principle of priority in taxonomic nomenclature, Tramea calverti Muttkowski, 1910 should hereafter be considered a junior synonym of Tramea darwini Kirby, 1889. Full article

Given the high medical importance of ticks, we analyzed the most common preventive behaviors and practices adopted by medical students from Poland, Germany, and Thailand, and the level of their knowledge of ticks and tick-borne diseases. A survey consisting of 19 questions was conducted among 636 randomly selected students. The study showed that the Polish and German students preferred inspection of the body on their return home (86.9% and 63.5%, respectively) and wearing protective clothes (79.8% and 32.3%, respectively) as part of prophylaxis. The Thai students most often chose wearing protective clothes (54.7%) and preventive behavior in tick habitats (42.7%). Approximately 7% of the Polish medical students and as many as 22% of the German and Thai respondents did not use any means of prevention. Our analyses suggest that the use of preventive methods and respondents’ behaviors depend on socio-demographic factors and the level of health education. The insufficient practical implementation of tick prevention measures by the medical students suggests a need for verification of health education programs in schools as well as effective popularization and educational activities. It is also necessary to develop a public health protection strategy against the effects of tick bites. Full article

The leaffooted bug, Leptoglossus zonatus (Heteroptera: Coreidae), has become a key pest of almonds, pistachios, and pomegranates in California. Adults and nymphs directly feed on nuts and fruits, which reduces crop yield and quality and can facilitate pathogen infections. Current monitoring strategies require growers to actively sample the tree canopy, with no economic thresholds being developed for this pest. To improve monitoring of L. zonatus, a three-year study was conducted to identify an optimal trap. A hanging cross-vane panel trap was identified as the best trap type in Year 1, and subsequent work in Years 1–3 focused on refining its use by modifying surface texture and color. Results indicated that coating trap surfaces with the lubricant fluon improved trap catching ability, and adults were most frequently recovered in yellow traps. A hanging cross-vane panel trap with these features could serve as the basis for the development of a new monitoring system for this pest in orchards, which could be improved further if semiochemical lures will be developed. Full article

We provide the first molecular phylogeny of Keroplatidae and Lygistorrhinidae, families of fungus gnats (Diptera: Bibionomorpha: Sciaroidea). Phylogenies reconstructed by Maximum Likelihood and Bayesian methods, based on four nuclear and four mitochondrial gene markers (5106 base pairs) sequenced for 75 genera and 105 species, show Keroplatidae as monophyletic only with the family Lygistorrhinidae included, herewith treated as the subfamily Lygistorrhininae stat. nov. The subfamily Arachnocampinae is retained in the family, although lowering its overall support. An early branching clade, comprising species of Platyura Meigen, 1803 and Paleoplatyura melanderi Fisher, 1941, forms subfamily Platyurinae Loew, 1850 stat. nov. The subfamilies Sciarokeroplatinae and Macrocerinae grouped together with three genera considered here as Keroplatidae incertae sedis. Subfamily Lygistorrhininae forms a sister clade to subfamily Keroplatinae, both retained monophyletic with high support. The traditional division of the subfamily Keroplatinae into the tribes Orfeliini and Keroplatini appears as outdated, resting largely on adaptive characters prone to parallel evolution. We find support for an alternative tribe corresponding to the Cloeophoromyia–Asindulum genus group, but a tribal reclassification of the Keroplatinae is left for future studies. The genus Heteropterna Skuse, 1888 is considered as identical with Ctenoceridion Matile, 1972 syn. nov. Full article

Red wood ants are keystone species of forest ecosystems in Europe. Environmental factors and habitat characteristics affect the size of their nest mounds, an important trait being in concordance with a colony’s well-being and impact on its surroundings. In this study, we investigated the effect of large-scale (latitude and altitude) and small-scale environmental factors (e.g., characteristics of the forest) on the size of nest mounds of Formica polyctena in Central Europe. We predicted that the change in nest size is in accordance with Bergmann’s rule that states that the body size of endotherm animals increases with the higher latitude and/or altitude. We found that the size of nests increased along the latitudinal gradient in accordance with Bergmann’s rule. The irradiation was the most important factor responsible for the changes in nest size, but temperature and local factors, like the perimeter of the trees and their distance from the nest, were also involved. Considering our results, we can better understand the long-term effects and consequences of the fast-changing environmental factors on this ecologically important group. This knowledge can contribute to the planning of forest management tactics in concordance with the assurance of the long-term survival of red wood ants. Full article

The thicknesses of the cuticle and rectal pads, and the spiracle morphology were compared for four termite species from different habitats, including one drywood termite, Cryptotermes brevis Walker, one “wetwood” termite, Cryptotermes cavifrons Banks, one subterranean termite, Coptotermes formosanus Shiraki, and one dampwood termite, Neotermes jouteli (Banks). Cuticle thicknesses were significantly different among all four termite species. Neotermes jouteli had the thickest cuticle, while Co. formosanus had the thinnest. The cuticle of C. brevis was thicker than that of C. cavifrons and Co. formosanus, which may reflect a comparably greater need to prevent water loss in drier habitats for C. brevis. Rectal pad widths were significantly different among all four termite species, except those of C. brevis with N. jouteli. The rectal pads of N. jouteli and C. brevis were thicker than those of C. cavifrons and Co. formosanus, and the rectal pads of C. cavifrons were thicker than those of Co. formosanus in turn. Larger rectal pads likely account for the water conservation mechanism of producing dry, pelleted frass in the kalotermitids (N. jouteli, C. brevis, and C. cavifrons). Morphological observations of the spiracles showed the presence of protuberances (atrial arms) in the three kalotermitids. The function of this protuberance is unclear, but it may serve as a sac-like structure, aiding in gas exchange, or a moisture trap aiding in the prevention of water loss through evaporation. Full article

Ticks are hematophagous ectoparasites of humans and animals. These arthropods employ different strategies in their host-seeking activity; most often, it is the “nest”- and “pasture-questing” behaviour. Some species, e.g., Dermacentor reticulatus, exhibit both types of activity depending on their developmental stage. The aim of the present study was to investigate the host-seeking activity of adult D. reticulatus ticks in the eastern part of Poland. To this end, ticks were collected with the flagging method during their seasonal activity in three different types of habitat. Active specimens were marked with a permanent marker and then released. This was repeated consistently at 7-day intervals using a different colour of the marker each time, which allowed tracking the questing activity of the specimens. Most frequently, repetitive tick activity (repeated up to seven times) was noted in a locality surrounded by urban developments. In an agriculturally unused open meadow habitat, 69.9% of D. reticulatus ticks were found to undertake questing activity only once. D. reticulatus females proved to be more aggressive and determined to find a host than the males of this species. Adult D. reticulatus ticks are able to stay in the habitat for a long time and undertake multiple host-seeking activities. The greatest threat of attacks on animals, including domestic animals, and sporadically humans, by these ticks occurs in meadow habitats, which are preferred by this species. Full article

Butterfly–host plant relationships can inform our understanding of ecological and trophic interactions that contribute to ecosystem function, resiliency, and services. The ecology of danaid–milkweed (Apocynaceae) host plant interactions has been studied in several biomes but is neglected in deserts. Our objective was to determine effects of plant traits, seasonality, and landscape-level host plant availability on selection of Mojave milkweed (Asclepias nyctaginifolia A. Gray) by ovipositing monarch butterflies (Danaus plexippus plexippus) and queen butterflies (Danaus gilippus thersippus) in the Californian Mojave Desert. We surveyed all known Mojave milkweed locations in the Ivanpah Valley, California (n = 419) during early, mid-, and late spring in 2017. For each survey, we counted monarch and queen butterfly eggs on each Mojave milkweed plant. We also measured canopy cover, height, volume, and reproductive stage of each Mojave milkweed plant. We counted a total of 276 queen butterfly eggs and zero monarch butterfly eggs on Mojave milkweed host plants. We determined that count of queen butterfly eggs significantly increased with increasing Mojave milkweed canopy cover. Additionally, count of queen butterfly eggs was: (1) greater on adult Mojave milkweed plants than on juvenile and seedling plants and greater on juvenile Mojave milkweed plants than on seedling plants; and (2) greater during early spring than mid-spring—we recorded no eggs during late spring. Based on aggregation indices, queen butterfly eggs occurred on Mojave milkweed plants in a nonrandom, clustered pattern throughout the Ivanpah Valley. We provide the first evidence of trophic interactions between queen butterflies and Mojave milkweed at multiple spatial scales in the Mojave Desert, suggesting that conservation and management practices for both species should be implemented concurrently. Given its role as an herbivore, pollinator and prey, the queen butterfly may serve as a model organism for understanding effects of anthropogenic disturbance (e.g., solar energy development) on “bottom-up” and trophic interactions among soils, plants and animals in desert ecosystems. Full article

The fall armyworm (Spodoptera frugiperda) is a pest of tropical origin which recently invaded Africa, the Far East and Australia. Temperature, therefore, plays an important role in its invasion biology, since this pest does not go into diapause. The aim of this study was to determine the development rate of S. frugiperda at different temperatures and to calculate the number of degree-days (°D) required for each stage to complete its development. This study was conducted at five different temperatures—18, 22, 26, 30 and 32 ± 1 °C. Larvae were reared individually in Petri dishes with sweetcorn kernels provided as food. The development rate of S. frugiperda increased linearly with increasing temperatures between 18 and 30 °C and larval survival was the highest between 26 and 30 °C. The optimal range for egg, larval and egg-to-adult development was between 26 and 30 °C. The optimum temperature with the fastest larval development rate and lowest mortality was at 30 °C. The pupal development period ranged between 7.82 and 30.68 days (32–18 °C). The minimum temperature threshold for egg and larva development was 13.01 and 12.12 °C, respectively, 13.06 °C for pupae and 12.57 °C for egg-to-adult development. Degree-day requirements for the development of the respective life cycle stages of S. frugiperda were 35.68 ± 0.22 for eggs, 204.60 ± 1.23 °D for larvae, 150.54 ± 0.93 °D for pupae and 391.61 ± 1.42 °D for egg-to-adult development. Full article

Management of the common bed bug (Cimex lectularius L.) necessitates the use of multiple control techniques. In addition to synthetic pesticides and mechanical interventions, plant-derived essential oils represent one of the control options. Mixtures of two or more essential oil components (monoterpenoids) exhibit synergistic toxicity effects against insects due to increased cuticular penetration. Monoterpenoids, such as carvacrol, eugenol and thymol, are neurologically active and inhibit the nerve firing activity of C. lectularius. However, the effects of mixtures of these monoterpenoids on their toxicity and neuroinhibitory potential against C. lectularius are not known. In this study, the toxicity levels of a tertiary mixture of carvacrol, eugenol and thymol (1:1:1 ratio) and a binary mixture of synthetic insecticides, bifenthrin and imidacloprid (1:1 ratio) were evaluated against C. lectularius through topical bioassays and electrophysiology experiments. Both a mixture of monoterpenoids and the mixture of synthetic insecticides exhibited synergistic effects in topical bioassays. In electrophysiology experiments, the monoterpenoid mixture led to greater neuroinhibitory effects, whereas a mixture of synthetic insecticides caused higher neuroexcitatory effects in comparison to single compounds. This study shows evidence for neurological mechanisms of synergistic interactions between monoterpenoids and provides information regarding the utilization of natural compound mixtures for C. lectularius management. Full article

The ant-loving beetle genus Panabachia Park 1942 is a poorly studied beetle lineage from the new world tropics. We recently collected Panabachia from several previously unrecorded locations in the páramo biome of the high Ecuadorian Andes, with males exhibiting great morphological variation in the distribution of the foveae and depressions in the pronotum, as well as aspects of the male genitalia. Here, we employ phylogenetic and species delimitation methods with mitochondrial (COI) and nuclear protein-coding (wingless) gene sequences to examine the concordance of morphological characters and geography with hypothesized species boundaries. Three methods of species delimitation (bPTP, GMYC and Stacey) were used to estimate the number of species, and divergence times between putative species using molecular clock calibration. Phylogenetic analysis revealed two parallel radiations, and species delimitation analyses suggest there are between 17 and 22 putative species. Based on clade support and concordance across species delimitation methods we hypothesize 17 distinct clusters, with allopatric speciation consistent with most geographic patterns. Additionally, a widespread species appears to be present in northern páramo sites, and some sister species sympatry may indicate other diversification processes have operated on certain lineages of Panabachia. Divergence time estimates suggest that Panabachia originated in the Miocene, but most species analyzed diverged during the Pliocene and Pleistocene (5.3–0.11 Mya), contemporaneous with the evolution of páramo plant species. Full article

Zoraptera is a small and predominantly tropical insect order with an unresolved higher classification due to the extremely uniform external body morphology. We, therefore, conducted a multigene molecular phylogeny of extant Zoraptera and critically re-evaluated their morphological characters in order to propose a natural infraordinal classification. We recovered a highly-resolved phylogeny with two main clades representing major evolutionary lineages in Zoraptera, for which we propose family ranks. The two families exhibit striking differences in male genitalia and reproductive strategies. Each family contains two subclades (subfamilies) supported by several morphological synapomorphies including the relative lengths of the basal antennomeres, the number and position of metatibial spurs, and the structure of male genitalia. The newly proposed higher classification of Zoraptera includes the family Zorotypidae stat. revid. with Zorotypinae Silvestri, 1913 (Zorotypus stat. revid., Usazoros Kukalova-Peck and Peck, 1993 stat. restit.) and Spermozorinae subfam. nov. (Spermozoros gen. nov.), and Spriralizoridae fam. nov. with Spiralizorinae subfam. nov. (Spiralizoros gen. nov., Scapulizoros gen. nov., Cordezoros gen. nov., Centrozoros Kukalova-Peck and Peck, 1993, stat. restit., Brazilozoros Kukalova-Peck and Peck, 1993, stat. restit.), and Latinozorinae subfam. nov. (Latinozoros Kukalova-Peck and Peck, 1993, stat. restit.). An identification key and morphological diagnoses for all supraspecific taxa are provided. Full article

(1) Intensive agriculture has a high impact on pollinating insects, and conservation strategies targeting agricultural landscapes may greatly contribute to their maintenance. The aim of this work was to quantify the effect that the vegetation of crop margins, with either herbaceous or shrubby plants, had on the abundance and diversity of bees in comparison to non-restored margins. (2) The work was carried out in an area of intensive agriculture in southern Spain. Bees were monitored visually and using pan traps, and floral resources were quantified in crop margins for two years. (3) An increase in the abundance and diversity of wild bees in restored margins was registered, compared to non-restored margins. Significant differences in the structure of bee communities were found between shrubby and herbaceous margins. Apis mellifera and mining bees were found to be more polylectic than wild Apidae and Megachilidae. The abundance of A. mellifera and mining bees was correlated to the total floral resources, in particular, to those offered by the Boraginaceae and Brassicaceae; wild Apidae and Megachilidae were associated with the Lamiaceae. (4) This work emphasises the importance of floral diversity and shrubby plants for the maintenance of rich bee communities in Mediterranean agricultural landscapes. Full article

The lone star tick (Amblyomma americanum L.) is native to the United States, with its primary range encompassing the Southeast and portions of the Midwest. It is an aggressive ectoparasite that actively seeks out hosts through detection of carbon dioxide and vibrations and can transfer ehrlichiosis-causing bacteria as well as a carbohydrate that causes alpha-gal syndrome (red meat allergy) in humans. It has become of increasing concern as its range has recently expanded into coastal regions of the Northeast. Historically, harsh northeastern winter weather conditions made these areas inhospitable for A. americanum survival, but a warming climate coupled with increased host availability seem to have facilitated their range expansion. We developed a study to observe the effects of weather conditions on adult A. americanum overwintering survival. The study was conducted over three years in Connecticut and Maine. Ground-level conditions were manipulated to determine the effects of differing combinations of natural insulative barriers (leaf litter and snow accumulation) on adult A. americanum survival. We determined that there was a significant difference in survival between the two states, between years in Maine, and between sexes within Connecticut. However, presence or absence of snow and/or leaf litter had no impact on survival. Overall, we found a positive correlation between mean hourly temperature and adult survival in Maine, where temperatures were consistently below freezing. The results of this study can be included in an adaptive, predictive analytic model to accommodate the expected fluctuations and range expansion of A. americanum that will most likely accompany an increase in temperatures throughout the Northeast. Full article

The invasive Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae), is the primary vector of the phloem-infecting bacterium, Candidatus Liberibacter asiaticus. Candidatus L. asiaticus is the putative causal agent of Huanglongbing (HLB) disease, a destructive disease of Citrus. While many Citrus species are susceptible to D. citri probing and HLB disease, there are marked behavioral differences in D. citri probing responses and Ca. Liberibacter asiaticus infection severity among Citrus species. Using four mandarin hybrid selections and pummelo plants variably resistant to D. citri probing, oviposition, and survival, we explored probing differences using electropenetrography (EPG), conducted an oviposition and survival study, and determined host plant metabolites using gas-chromatography mass-spectroscopy (GC-MS). We found thirty-seven D. citri probing variables to be significantly different among tested mandarin selections and pummelo, in addition to differential oviposition and survivorship abilities on tested plants. We found sixty-three leaf metabolites with eight being significantly different among tested mandarin selections and pummelo. Detailed analysis of probing behavior, oviposition, survivorship, and host plant metabolite concentrations reveals the complex, layered resistance mechanisms utilized by resistant Citrus against D. citri probing. EPG is a powerful technology for screening Asian citrus psyllid resistant Citrus to elucidate host plant-vector interactions, with an aim to minimize vector probing and eliminate the spread of the bacterial pathogen, Ca. L. asiaticus. Full article

A push-pull strategy for reducing populations of the thrips Frankliniella occidentalis (Pergande), F. bispinosa (Morgan) and F. tritici (Fitch) in tomato was evaluated. Push components consisted of ultraviolet (UV)-reflective mulch and foliar applications of kaolin and the pull component consisted of the companion plant Bidens alba (L.). Replicated field experiments were conducted in 2011 and 2012. Adult and larval thrips were reduced by UV-reflective mulch during early and mid-flowering of tomato. Spray applications of kaolin were effective in reducing adult and larval thrips during early, mid- and late-flowering. The pull effects of the B. alba companion plants were additive and sometimes interactive with the push effects of UV-reflective mulch and kaolin in reducing the adult males of each thrips species and the females of F. bispinosa. The strategy was not effective in reducing the adult females of F. tritici and F. occidentalis. In addition to attracting the Frankliniella species adults, the companion plants were hosts for the thrips predator Orius insidiosus (Say). The companion plants combined with UV-reflective mulch and kaolin proved effective as a push-pull system for suppressing flower thrips, including F. occidentalis which is a serious pest of tomato worldwide. Full article

Emerging pests are increasingly threatening fruit orchard health across the Mediterranean area. Tephritidae, representing serious threats for Europe, are numerous, and the fruit flies Bactrocera zonata and those belonging to Bactrocera dorsalis complex are among the most alarming species. These species are highly polyphagous and B. zonata has already spread to some Mediterranean countries. Due to these ongoing threats, in the Campania Region (southern Italy), a survey with traps and infested fruits analysis was performed with the aim of detecting the presence of species of Bactrocera dorsalis complex. In two mixed fruit-trees fields, some adults belonging to a species of Bactrocera were captured in traps baited with the highly attractive male lure (methyl eugenol). They were distinguished from similar-looking Bactrocera spp. by morphological and molecular comparative analyses. Considering the existing morphological keys, specimens were tentatively identified as B. dorsalis but molecular characterization with COI split them into two clades. Some specimens were grouped with B. dorsalis similar to B. kandiensis and B. kandiensis and others in a clade including B. dorsalis and B. invadens (syn. B. dorsalis). ITS1 sequences instead confirmed morphological identification. The integrative approach allowed identifying all the specimens collected as belonging to the B. dorsalis complex. This finding represents the first field interception in Europe of a member of one of the most dangerous groups of fruit flies. Full article

The phytosanitary emergency caused by the spread of Xylella fastidiosa in the Mediterranean has raised demands for a better understanding of the ecology of its presumed and candidate insect vectors. Here, we present the results of a two-year survey carried out in olive groves across southern, eastern and Central Spain and northeastern Portugal. Several sampling methods were tested and compared to select the most appropriate to estimate population levels of potential vectors of X. fastidiosa. The spittlebugs Philaenus spumarius and Neophilaenus campestris (Hemiptera: Aphrophoridae) were the main species associated with olive groves. Both species were widely present on herbaceous ground vegetation within the olive groves; P. spumarius mainly associated with Asteraceae and N. campestris with Poaceae. Due to the patchy distribution of spittlebugs within the olive groves, sweep nets were the most effective and least time-consuming sampling method for the estimation of population size both in the ground cover and tree canopies. Trends in population density showed that spittlebugs can be abundant on ground vegetation but very rare on olive canopies. Spittlebugs disperse in late spring to non-cultivated hosts that act as natural reservoirs. In late fall, adults return to the olive groves for oviposition. However, olive trees may act as transient hosts for spittlebugs and high population densities of these insect vectors should be avoided in areas where X. fastidiosa is present. Full article

In urban settings, chikungunya, Zika, and dengue viruses are transmitted by Aedes aegypti mosquitoes. Since these viruses co-circulate in several regions, coinfection in humans and vectors may occur, and human coinfections have been frequently reported. Yet, little is known about the molecular aspects of virus interactions within hosts and how they contribute to arbovirus transmission dynamics. We have previously shown that Aedes aegypti exposed to chikungunya and Zika viruses in the same blood meal can become coinfected and transmit both viruses simultaneously. However, mosquitoes may also become coinfected by multiple, sequential feeds on single infected hosts. Therefore, we tested whether sequential infection with chikungunya and Zika viruses impacts mosquito vector competence. We exposed Ae. aegypti mosquitoes first to one virus and 7 days later to the other virus and compared infection, dissemination, and transmission rates between sequentially and single infected groups. We found that coinfection rates were high after sequential exposure and that mosquitoes were able to co-transmit both viruses. Surprisingly, chikungunya virus coinfection enhanced Zika virus transmission 7 days after the second blood meal. Our data demonstrate heterologous arbovirus synergism within mosquitoes, by unknown mechanisms, leading to enhancement of transmission under certain conditions. Full article

California strawberries have two major arthropod pests—the twospotted spider mite, Tetranychus urticae and the western tarnished plant bug, Lygus hesperus, which result in significant losses to the yield and quality of marketable berries. Other important insect pests that are frequently seen in strawberry include the greenhouse whitefly, Trialeurodes vaporariorum and the western flower thrips, Frankliniella occidentalis that cause varying levels of damage depending on the level of infestation. Chemical pesticides play a major role in managing these pests but not without the associated risk of pesticide resistance and environmental safety. Two field studies were conducted in commercial strawberry fields in Santa Maria, one of the strawberry growing areas in California Central Coast, to determine the efficacy of chemical, botanical and microbial pesticides in the integrated pest management (IPM) of strawberry. Chemical, botanical and microbial pesticides were evaluated against T. urticae in a small plot study in 2013 and against L. hesperus and other insect pests in a large plot study in 2015 in commercial strawberry fields. Bug vacuums were also used in the 2015 study. Results demonstrated that non-chemical alternatives can play an important role in strawberry IPM. Full article

The mechanisms allowing the widespread invasive pest Drosophila suzukii to survive from early spring until the availability of the first fruit crops are still unclear. Seasonal biology and population dynamics of D. suzukii were investigated in order to better understand the contribution of the early spring hosts to the infestation of the first fruit crops of the season. We identified hosts available to D. suzukii in early spring and assessed their suitability for the pest oviposition and reproductive success under field and laboratory conditions. The natural infestation rate of one of these hosts, Aucuba japonica, was assessed over springtime and the morphology of the flies that emerged from infested A. japonica fruits was characterized under field conditions. Then, these findings were correlated with long-term monitoring data on seasonal reproductive biology and morphology of the pest, using a cumulative degree-days (DD) analysis. Field sampling revealed that overwintered D. suzukii females were physiologically able to lay eggs at 87 DD which coincided with the detection of the first infested early spring hosts. The latter were continuously and increasingly infested by D. suzukii eggs in nature from early spring until the end of May, in particular Aucuba japonica. Individuals emerged from most of these hosts were characterized by a poor fitness and a rather low success of emergence. In the field, only few summer morphs emerged from naturally infested A. japonica fruits around the end of May-beginning of June. However, field monitoring in orchards revealed that D. suzukii individuals consisted solely of winter morphs until mid-June. These observations indicate that overwintered D. suzukii females are the predominant source for the infestations in the first available fruit crops of the season. We discuss these findings in the context of possible pest control strategies. Full article

Mosquito-borne diseases account for the deaths of approximately 700,000 people annually throughout the world, with many more succumbing to the debilitating side effects associated with these etiologic disease agents. This is exacerbated in many countries where the lack of mosquito control and resources to prevent and treat mosquito-borne disease coincide. As populations of mosquito species grow more resistant to currently utilized control chemistries, the need for new and effective chemical means for vector control is more important than ever. Previous work revealed that plant essential oils enhance the toxicity of permethrin against multiple mosquito species that are of particular importance to public health. In this study, we screened permethrin and deltamethrin in combination with plant essential oils against a pyrethroid-susceptible and a pyrethroid-resistant strain of both Aedes aegypti and Anopheles gambiae. A number of plant essential oils significantly enhanced the toxicity of pyrethroids equal to or better than piperonyl butoxide, a commonly used synthetic synergist, in all strains tested. Significant synergism of pyrethroids was also observed for specific combinations of plant essential oils and pyrethroids. Moreover, plant essential oils significantly inhibited both cytochrome P450 and glutathione S-transferase activities, suggesting that the inhibition of detoxification contributes to the enhancement or synergism of plant essential oils for pyrethroids. This study highlights the potential of using diverse plant oils as insecticide additives to augment the efficacy of insecticidal formulations. Full article

Many methods for trajectory simulation, such as Hybrid Single-Particle Lagrangian Integrated Trajectory (HYSPLIT), have been developed over the past several decades and contributed greatly to our knowledge in insect migratory movement. To improve the accuracy of trajectory simulation, we developed a new numerical trajectory model, in which the self-powered flight behaviors of insects are considered and trajectory calculation is driven by high spatio-temporal resolution weather conditions simulated by the Weather Research and Forecasting (WRF) model. However, a rigorous evaluation of the accuracy of different trajectory models on simulated long-distance migration is lacking. Hence, in this study our trajectory model was evaluated by a migration event of the corn earworm moth, Helicoverpa zea, in Texas, USA on 20–22 March 1995. The results indicate that the simulated migration trajectories are in good agreement with occurrences of all pollen-marked male H. zea immigrants monitored in pheromone traps. Statistical comparisons in the present study suggest that our model performed better than the popularly-used HYSPLIT model in simulating migration trajectories of H. zea. This study also shows the importance of high-resolution atmospheric data and a full understanding of migration behaviors to the computational design of models that simulate migration trajectories of highly-flying insects. Full article