Morphology of male genitalia of culicids is generally species-specific and often used as a taxonomic marker. However, some characters of the male genitalia vary intraspecifically and are not taxonomically diagnostic. This might be the case of Aedes scapularis, a Neotropical culicid with vector competence for arboviruses and filarial worms. Males of this species may or not present a retrorse process (RP) in the genitalic claspette filaments, which led authors to suspect that this variance might be indicative of population divergence or incipient speciation process. This suspicion has not been investigated hitherto and it is not known if there are variable patterns of RPs. We hypothesized that the presence of the RP varies intraspecifically in Ae. scapularis and then we statistically evaluated the variability of this character in a single population. To this study the genitalia of 73 males of Ae. scapularis were prepared, and their RPs were meristically quantified and categorized according to the phenotypes observed. We noted that the presence or RPs is a polymorphic character because it varied inter and intra-individually. The presence of a single RP on each claspette filament was the predominant pattern (77%), but absent or multiple RPs in each filament were also found either in bilateral symmetry or asymmetry. Thus, we conclude that the presence of RPs owing to its high variability is not indicative of populational divergence or diagnostic of species complex within Ae. scapularis.

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.

Fungus-growing termites (Macrotermitinae) are important pests in tropical countries. They are difficult to control with existing baiting methods, as chitin synthesis inhibitors are not effectual as active ingredients. We tested two neurotoxins, fipronil and imidacloprid, as potential bait active ingredients against Macrotermes gilvus (Hagen) in Singapore. In laboratory bioassays, M. gilvus showed no preference for doses of 0–64 ppm fipronil, or for doses of 0–250 ppm imidacloprid, indicating no repellence. We tested each insecticide in toilet paper as a bait matrix in a field experiment. After 28 days, termites had eaten 5–13% of the fipronil treated toilet paper, abandoned bait and monitoring stations, contacted no new stations, and repaired poorly their experimentally damaged mounds. Termites ate no imidacloprid treated toilet paper, abandoned bait stations although contacted new stations, and repaired fully their damaged mounds. Termites ate 60–70% of the control toilet paper, remained in bait stations, and fully repaired damaged mounds. After 56 days, all five fipronil colonies were eliminated, whereas all of the imidacloprid and control colonies were healthy. The results suggest that fipronil could be an effective active ingredient in bait systems for fungus-growing termites in tropical countries.

Mite growth inhibitors (MGIs), such as etoxazole and hexythiazox, are valuable IPM tools for Tetranychus urticae control in hops due to their unique mode of action and selectivity. Hence, it is necessary to standardize bioassay methods to evaluate the efficacy of MGIs, monitor resistance, and identify mechanisms underlying MGI resistance in the field. Here, we developed a three-tiered approach for evaluating ovicidal toxicity of MGIs to T. urticae, which simulated different MGI exposure scenarios in the field. The most effective bioassay method was direct exposure of T. urticae eggs to MGIs. With this method, four field-collected T. urticae populations showed low-to-moderate resistance to MGIs. Cross-resistance among MGIs and from MGIs to bifenazate and bifenthrin was detected. Besides target site insensitivity, enhanced cytochrome P450 and esterase activities also contribute to the MGI resistance in hop yard-collected T. urticae populations. Low-to-moderate MGI resistance in T. urticae populations may be mediated by multiple mechanisms. Positive selection pressure on the I1017F mutation is moderate in field-collected T. urticae populations. Further studies are required to identify metabolic detoxification genes that confer resistance to MGIs for precise resistance monitoring.

There are several possible causes of aggregated nesting in solitary Aculeata, one being joint defense against parasites. We tested whether females prefer nesting in aggregations, even if they consist of heterospecifics. We compared the colonization and nesting parasitism of trap-nests with and without a red mason bee aggregation. The results did not support our hypothesis that females prefer nesting in aggregations. The numbers of wild Aculeata nests did not differ between trap-nests with and without an aggregation. Unexpectedly, parasitism rates were higher in trap-nests with aggregations. When analyzing only nests of wild insects (mostly wasps), the differences in parasitism disappeared. Natural nesting sites may be such a limited resource that females nested in the first trap-nest they encountered and did not discriminate between our treatments, or wasps might share too few parasites species with bees to benefit from joint nest defense.

Olfaction is a key sense routing foraging behaviour in parasitoids. Preferences for food, mate and host stimuli can be innate in parasitic wasps. Alternatively, learning-mediated mechanisms play a crucial role. Females of the braconid parasitoid Psyttalia concolor exploit olfactory cues arising from tephritid hosts and related microhabitats. However, little is known on the olfactory stimuli routing males searching for mates. In this study, we focused on the attractiveness of Bactrocera oleae-induced olive volatiles towards P. concolor males. Furthermore, we evaluated learning occurrence in virgin males, when trained for selected unattractive volatile organic compounds (VOCs) associated with mate rewards. (E)-β-Ocimene, α-pinene and limonene attracted virgin males in Y-tube bioassays. Unattractive VOCs evoked positive chemotaxis after associative learning training. P. concolor males exposed to VOCs during a successful or unsuccessful mating, showed short-term preference for these VOCs (<1 h). However, memory consolidation was strictly dependent on reward value. Indeed, males experiencing a successful mating showed a fast consolidation into protein dependent long-term memory, appearing after 24 h. On the other hand, males experiencing a less valuable training experience (i.e. unsuccessful courtship), did not show consolidated memory after 24 h. Overall, our findings suggest that P. concolor virgin males may exploit VOCs from the host microhabitat to boost their mate searching activity, thus their reproductive success. However, since learning is a costly process, P. concolor males retained durable memories just in presence of a valuable reward, thus avoiding maladaptive behaviours.

The flee-weevil Orchestes steppensis Korotyaev (Coleoptera: Curculionidae) is a steppe eastern Palaearctic species, notable as a serious pest of elms (Ulmus spp., Ulmaceae), by feeding on the leaves (adults) or mining them heavily (larvae), especially of Ulmus pumila L. in Xinjiang, China. We have corrected the previous misidentifications of this weevil in China as O. alni (L.) or O. mutabilis Boheman and demonstrated that it is likely to be an invasive species in Xinjiang. Prior to this study, natural enemies of O. steppensis were unknown in Xinjiang. Resulting from field investigation and rearing in the laboratory during 2013–2016, seven parasitoid species were found to be primary and solitary, attacking larval and pupal stages of the host weevil. Pteromalus sp. 2 is the dominant species and also is the most competitive among the seven parasitoids, which could considered to be a perspective biological control agent of O. steppensis. Yet, the current control of this pest by the local natural enemies in Xinjiang is still currently inefficient, even though in 2016 parasitism was about 36% on U. pumila in Urumqi, so the potential for a classical biological control program against it needs to be further investigated, including an assessment of its parasitoids and other natural enemies in the native range of O. steppensis. The presented information on the natural enemies of this weevil can be also important for a potential classical biological control program against it in North America (Canada and USA), where it is a highly damaging and rapidly spreading invasive species.

The lesser date moth (LDM) Batrachedra amydraula is a significant pest of date palm fruits. Previously, detection and monitoring of the pest was inaccurate due to high costs of sampling with lifting machines. We report a practical system for detection and monitoring of LDM based on pheromone traps and relevant models. Dose–response experiments with LDM pheromone traps indicated a 1 mg lure is optimal for monitoring. Delta traps with adhesive covering their entire inner surface gave the highest captures while trap colour was unimportant. Sampling pheromone traps throughout the night indicated male flight began at 1:00–2:00 and reached a peak 2 h before sunrise. Monitoring traps exposed all year long in Israel revealed three generations with different abundance. Trapping transects in a date plantation indicated interference from a monitoring trap became minimal at distances >27 m away. Inter-trap distances closer than this may lower efficiency of monitoring and mass trapping in control programs. Our estimate of the circular effective attraction radius (EARc) of a 1 mg delta trap for LDM (3.43 m) shows this bait is among the most attractive compared with baits for other insects. We developed encounter-rate equations with the pheromone trap EARc to model the interplay between population levels, trap density and captures that are useful for detection of invasive LDM and its control by mass trapping. The integrated methodologies are applicable to many pest species.

Tetraneura ulmi (L.), a member of Eriosomatinae subfamily, is one of the gall-forming aphids occurring on elms. Sap-sucking behaviour of founding mothers results in the formation of new plant organs. This study documents the changes in the content of plant biogenic amines (putrescine, cadaverine, spermidine, tryptamine, spermine and histamine) and key enzymes of their biosynthesis: lysine decarboxylase (LDC), tyrosine decarboxylase and ornithine decarboxylase (ODC) in galls and other parts of Siberian elm (Ulmus pumila L.) leaves during the galling process. The direction and intensity of these changes for particular amines and enzymes were dependent on the stage of gall development and part of the galling leaf. Generally, the amine content tended to increase in gall tissues during the 1st and 2nd period of the galling process and decreased in later phases. LDC and ODC activities were markedly enhanced, especially in gall tissues at the initial stage of the galling process.

Cytochrome P450s (CYPs or P450s) have been long recognized as very important enzymes in the metabolism of xenobiotic and endogenous compounds, but only a few CYPs have been functionally characterized in insects. The effort in functional characterization of insect P450s is heavily hindered by technical difficulties in preparing active, individual P450 enzymes directly from the target insect. In this paper, we describe the functional expression of two additional pyrethroid resistance-associated CYP9A genes (CYP9A12 and CYP9A17) from the polyphagous pest Helicoverpa armigera in the facile Escherichia coli. The functionality of E. coli produced CYP9A12, CYP9A14, and CYP9A17 was investigated and activities of these CYP9As were compared against three probe substrates after reconstitution with NADPH-dependent cytochrome P450 reductase. The results showed that active forms of CYP9A12 and CYP9A17 were expressed in E. coli with a content of about 1.0–1.5 nmol mg−1 protein in membrane preparations. In vitro assays showed that CYP9A14 was capable of catalyzing O-dealkylation of methoxyresorufin (MROD), ethoxyresorufin (EROD), and benzyloxyresorufin (BROD), while CYP9A12 and CYP9A17 exhibited only MROD and EROD activities. Kinetic studies demonstrated that CYP9A14 had the greatest kcat/Km value for MROD, and CYP9A17 for EROD, while the lowest kcat/Km values for both MROD and EROD were observed for CYP9A12. The distinct biochemical traits suggest that the three paralogous CYP9As may play different roles in xenobiotic metabolism in this important pest.

Thiamine is a vitamin that has been shown to act as a trigger to activate plant defence and reduce pathogen and nematode infection as well as aphid settling and reproduction. We have here investigated whether thiamine treatments of seeds (i.e. seed dressing) would increase plant resistance against aphids and whether this would have different effects on a generalist than on specialist aphids. Seeds of wheat, barley, oat and pea were treated with thiamine alone or in combination with the biocontrol bacteria Pseudomonas chlororaphis MA 342 (MA 342). Plants were grown in climate chambers. The effects of seed treatment on fecundity, host acceptance and life span were studied on specialist aphids bird cherry-oat aphid (Rhopalosiphum padi L.) and pea aphid (Acyrthosiphon pisum Harris) and on the generalist green peach aphid (Myzus persicae, Sulzer). Thiamine seed treatments reduced reproduction and host acceptance of all three aphid species. The number of days to reproduction, the length of the reproductive life, the fecundity and the intrinsic rate of increase were found reduced for bird cherry-oat aphid after thiamine treatment of the cereal seeds. MA 342 did not have any effect in any of the plant-aphid combinations, except a weak decrease of pea aphid reproduction on pea. The results show that there are no differential effects of either thiamine or MA 342 seed treatments on specialist and generalist aphids and suggest that seed treatments with thiamine has a potential in aphid pest management.

The Red Palm Weevil (RPW) Rhynchophorus ferrugineus (Olivier 1790) is an invasive pest from southeastern Asia and Melanesia that in the last 30 years has spread widely in the Middle East and Mediterranean Basin. Its stem-boring larvae cause great damage to several palm species of the Arecaceae family, many of which are economically important for agricultural and ornamental purposes. Therefore, great attention has recently been focused in studying this species to identify sustainable and effective eradication strategies, such as sterile insect technique (SIT). The rapid spread of RPW is associated with its high reproductive success. To evaluate the suitability of a SIT strategy, particular physiological and behavioral aspects of RPW reproduction, such as the presence of polyandry and post-copulatory sperm selection mechanisms, were investigated. To determine paternity of progeny from multiply mated females, double-crossing experiments were carried out confining individual females with either a wild-type male or a γ-irradiated male (Co-60). Fecundity and fertility of females were scored to evaluate post-copulatory sperm selection. Results showed that progeny were almost exclusively produced by the sperm of the second male, suggesting that a last-male sperm precedence is expressed at high levels in this species, and providing interesting insights for an area-wide RPW management strategy such as the SIT.

The aim of this study was to verify the occurrence of the coffee berry borer, Hypothenemus hampei (Ferrari), and to evaluate the population fluctuation of the pest in the Brazilian Cerrado (Federal District). The study was conducted, between November 2014 and October 2015, at Embrapa Cerrados (Planaltina/DF, Brazil) in an irrigated conilon coffee production area. In November 2014, 120 samples (ten berries/sample) were collected from berries that had fallen on the ground from the previous harvest. Between November 2014 and October 2015, insects were collected weekly, using traps (polyethylene terephthalate bottles) baited with ethyl alcohol (98 GL), ethyl alcohol (98 GL) with coffee powder, or molasses. Between January and July 2015, samples were collected fortnightly from 92 plants (12 berries per plant). All samples were evaluated for the presence of adult coffee berry borers. Samples from the previous harvest had an attack incidence of 72.4%. The baited traps captured 4062 H. hampei adults, and showed no statistical difference in capture efficiency among the baits. Pest population peaked in the dry season, with the largest percentage of captured adults occurring in July (31.0%). An average of 18.6% of the collected berries was attacked by the borer and the highest percentage incidence was recorded in July (33.2%). Our results suggest that the coffee berry borer, if not properly managed, could constitute a limiting factor for conilon coffee production in the Brazilian Cerrado.

Helicoverpa gelotopoeon, the South American bollworm, is a polyphagous pest of the Heliothinae complex that causes damage to soybean, cotton, and chickpea crops. Some species within this complex have developed resistance to genetically modified crops and insecticides, which has led to increased interest in their genetic diversity and population structure. The objective of this study was to characterize biological and reproductive parameters of two populations of H. gelotopoeon collected in two different provinces of Argentina. Intra- and inter-population crosses revealed that H. gelotopoeon populations from both regions of Argentina did not present evidence of pre-zygotic and post-zygotic incompatibility, suggesting that Tucumán and Córdoba populations of H. gelotopoeon belong to a single wide-ranging species. Our data support the occurrence of substantial gene flow between H. gelotopoeon populations, probably due to the widely documented, long-range migratory capacity of Heliothinae species.

The Artocarpus fruit fly, Bactrocera umbrosa (Fabricius) (Diptera: Tephritidae), is an oligophagous fruit pest infesting Moraceae fruits, including jackfruit (Artocarpus heterophyllus Lamarck), a fruit commodity of high value in Malaysia. The scarcity of fundamental biological, physiological and ecological information on this pest, particularly in relation to behavioural response to phytochemical lures, which are instrumental to the success of many area-wide fruit fly control and management programmes, underpins the need for studies on this much-underrated pest. The positive response of B. umbrosa males to methyl eugenol (ME), a highly potent phytochemical lure, which attracts mainly males of many Bactrocera species, was shown to increase with increasing age. As early as 7 days after emergence (DAE), ca. 22% of males had responded to ME and over 50% by 10 DAE, despite no occurrence of matings (i.e. the males were still sexually immature). Male attraction to ME peaked from 10 to 27 DAE, which corresponded with the flies’ attainment of sexual maturity. In wind-tunnel assays during the dusk courtship period, ME-fed males exhibited earlier calling activity and attracted a significantly higher percentage of virgin females compared with ME-deprived males. ME-fed males enjoyed a higher mating success than ME-deprived males at 1-day post ME feeding in semi-field assays. ME consumption also promotes aggregation behaviour in B. umbrosa males, as demonstrated in wind-tunnel and semi-field assays. We suggest that ME plays a prominent role in promoting sexual communication and enhancing mating performance of the Artocarpus fruit fly, a finding that is congruent with previous reports on the consequences of ME acquisition by other economically important Bactrocera species.

Ceratovacuna lanigera Zehntner is a major leaf pest of sugarcane. Widely distributed, it affects both the yield and quality of sugarcane in China. This study aimed to assess real yield and sugar yield losses, and the effect of C. lanigera damage on emergence of newly planted and ratoon cane under current production levels. Field experiments were carried out from 2014 to 2016 in Yunnan Province China. At maturity, plants were harvested and weighed to determine yield, and the effect on sugarcane quality and sucrose content analyzed. Real yield decreased by average of 46,185 kg hm−2 (range: 37,545–61,845 kg hm−2) in damaged versus undamaged areas, with an average yield loss rate of 35.9% (28.5–45.7%). Juice yield decreased by an average of 3.01% (2.4–4.13%) and sucrose content by 6.38% (5.48–8.16%). Juice brix decreased by an average of 7.66°BX (6.95–9.05°BX) and juice gravity purity by 12.35% (8.43–19.97%). In contrast, the reducing sugar content increased by an average of 1.21% (1.01–1.3%). Emergence rates of newly planted cane decreased by an average of 26.0% (24.7–27.3%). The emergence number of ratoon cane decreased by 66,834 hm2 (57,429–76,238 hm−2) and relative emergence loss rates of ratoon cane decreased by an average of 57.8% (57.6–58.0%). These findings confirm that C. lanigera damage severely affects sugarcane yield and quality in Yunnan Province. The results will help the implementation of effective control measures, thereby supporting sustainable development of the Chinese sugar industry.

Disentangling trophic interactions among species is important for elucidating mechanisms underlying ecosystem functioning and services. Carabid beetles are an important guild of predators that may regulate pest populations in arable landscapes, but their generalist feeding behavior hinders predictions about their actual contribution to pest control. In order to assess carabids’ potential for pest control, we simultaneously analyzed the carbon and nitrogen stable isotope ratios of a community of 45 co-occurring species in wheat and oilseed rape fields. With the expectation to identify distinct trophic groups based on the mean and the variance of carabid isotopic signatures, we observed a high degree of overlap in trophic positions between species. However, we also observed that species could be successfully categorized into two groups according to whether or not their carbon signatures varied independently from variations in the crop baseline. We interpret these results as differential primary resource uptake or by differential mobility aptitude in foraging. Accordingly, we propose that the isotopic signal can inform us on the presence/absence of links between generalist predators and cultivated plants through the trophic networks they belong to, and consequently on their potential role as pest natural enemies. We therefore suggest the complementarity of stable isotope analysis for obtaining a time-integrated assessment of carabid trophic behavior that may be combined with more direct molecular diet analysis allowing the simultaneous quantification of specific trophic links within agricultural landscapes.