Aphids are important agricultural pests worldwide. Their control is largely based on chemical insecticides. One species that shows important invasive abilities and host-plant-related differences is Therioaphis trifolii (Monell) (Hemiptera: Aphididae). T. trifolii maculata, also known as spotted alfalfa aphid (SAA), can be very injurious to alfalfa crops in certain regions, such as in Saudi Arabia for effective control it is essential to diagnose and monitor the resistance mechanisms in the SAA populations. In the present study, we analysed acetylcholinesterase (ace) target site insensitivity mechanisms. A 650 bp length DNA containing the putative acetylcholinesterase (ace1) precursor was obtained and compared with other Hemipteran species. The sequences of many individual aphids collected from alfalfa crops in Saudi Arabia were analysed for the presence of resistance mutations: no resistance mutations were found at the resistance mutation loci 302; however, the presence of a serine–phenylalanine substitution (S431F) was identified in one individual. The S431F substitution, has been shown to confer significant levels of both organophosphate and carbamate resistance in other aphid species, and is now found for the first time in T. trifolii. We subsequently developed a simple polymerase chain reaction–restriction fragment length polymorphism assays for the S431F mutation, using a TaqI restriction site destroyed by the S431F mutation. The novel diagnostic assay may support the implementation of Insecticide Resistance Management strategies, for the control of SAA in alfalfa crops in the Kingdom of Saudi Arabia, and other countries worldwide.

Flat grain beetles of the genus Cryptolestes (Coleoptera: Laemophloeidae) are one of the economically most important stored-product pests which feed on many kinds of agricultural products, especially grains. Nine of more than 40 described Cryptolestes species are recognized as stored-product pests and two of the pest species have a cosmopolitan distribution. Given the rapid growth in global trade of food products, ecological barriers to the spread of pests are easily overcome. Therefore, development of reliable systems for routine quarantine inspection and early infestation detection is vital. In the present study, we established a new rapid and accurate cytochrome c oxidase subunit I-based system for molecular identification of five common stored-product Cryptolestes species, namely, Cryptolestes capensis, Cryptolestes ferrugineus, Cryptolestes pusilloides, Cryptolestes pusillus and Cryptolestes turcicus. Five species-specific primer pairs for traditional uniplex polymerase chain reaction assay are described and their specificity and sensitivity for the identification process is evaluated using larval samples of 12 different populations from three continents (Asia, Europe and North America).

Metabarcoding, the coupling of DNA-based species identification and high-throughput sequencing, offers enormous promise for arthropod biodiversity studies but factors such as cost, speed and ease-of-use of bioinformatic pipelines, crucial for making the leapt from demonstration studies to a real-world application, have not yet been adequately addressed. Here, four published and one newly designed primer sets were tested across a diverse set of 80 arthropod species, representing 11 orders, to establish optimal protocols for Illumina-based metabarcoding of tropical Malaise trap samples. Two primer sets which showed the highest amplification success with individual specimen polymerase chain reaction (PCR, 98%) were used for bulk PCR and Illumina MiSeq sequencing. The sequencing outputs were subjected to both manual and simple metagenomics quality control and filtering pipelines. We obtained acceptable detection rates after bulk PCR and high-throughput sequencing (80–90% of input species) but analyses were complicated by putative heteroplasmic sequences and contamination. The manual pipeline produced similar or better outputs to the simple metagenomics pipeline (1.4 compared with 0.5 expected:unexpected Operational Taxonomic Units). Our study suggests that metabarcoding is slowly becoming as cheap, fast and easy as conventional DNA barcoding, and that Malaise trap metabarcoding may soon fulfill its potential, providing a thermometer for biodiversity.

Understanding the impacts of weather fluctuations, and environmental gradients, on the abundance of vectors is fundamental to grasp the dynamic nature of the entomological risk for disease transmission. The mosquito Armigeres subalbatus (Coquillet) is a common vector of filariasis. Nevertheless, its population dynamics have been relatively poorly studied. Here, we present results from a season long study where we studied spatio-temporal abundance patterns of Ar. subalbatus across the altitudinal gradient of Mt. Konpira in Nagasaki, Japan. Spatially, we found that abundance of adult Ar. subalbatus decreased with altitude and increased in areas where the ground was rich in leaf litter. Similarly, adult activity was observed only when relative humidity was over 65%. Temporally, we found that peaks in abundance followed large rainfall events. Nevertheless, this mosquito was under significant density dependence regulation. Our results suggest that Ar. subalbatus population peaks following large rainfall events could reflect the recruitment of individuals that were dormant as dry eggs. We did not find a clear signal of temperature on abundance changes of this mosquito, but only on its phenology. Since ground cover seemed more critical than temperature to its spatial distribution, we propose that this mosquito might have some degree of autonomy to changes in temperature.

Grasshopper plagues have seriously disturbed grassland ecosystems in Inner Mongolia, China. The accurate prediction of grasshopper infestations and control of grasshopper plagues have become urgent needs. We sampled 234, 342, 335, and 369 plots in Xianghuangqi County of Xilingol League in 2010, 2011, 2012, and 2013, respectively, and measured the density of the most dominant grasshopper species, Oedaleus decorus asiaticus, and the latitude, longitude, and associated relatively stable habitat factors at each plot. We used Excel–GeogDetector software to explore the effects of individual habitat factors and the two-factor interactions on grasshopper density. We estimated the membership of each grasshopper density rank and determined the weights of each habitat category. These results were used to construct a model system evaluating grasshopper habitat suitability. The results showed that our evaluation system was reliable and the fuzzy evaluation scores of grasshopper habitat suitability were good indicators of potential occurrence of grasshoppers. The effects of the two-factor interactions on grasshopper density were greater than the effects of any individual factors. O. d. asiaticus was most likely to be found at elevations of 1300–1400 m, flat terrain or slopes of 4–6°, typical chestnut soil with 70–80% sand content in the top 5 cm of soil, and medium-coverage grassland. The species preferred temperate bunchgrass steppe dominated by Stipa krylovii and Cleistogenes squarrosa. These findings may be used to improve models to predict grasshopper occurrence and to develop management guidelines to control grasshopper plagues by changing habitats.

The leafminer Coptodisca sp. (Lepidoptera: Heliozelidae), recently recorded for the first time in Europe on Italian black and common walnut trees, is shown to be the North-American Coptodisca lucifluella (Clemens) based on morphological (forewing pattern) and molecular (cytochrome oxidase c subunit I sequence) evidence. The phylogenetic relatedness of three species feeding on Juglandaceae suggests that C. lucifluella has likely shifted, within the same host plant family, from its original North-American hosts Carya spp. to Juglans spp. Over the few years since its detection, it has established in many regions in Italy and has become a widespread and dominant invasive species. The leafminer completes three to four generations per year, with the first adults emerging in April–May and mature larvae of the last generation starting hibernation in September–October. Although a high larval mortality was recorded in field observations (up to 74%), the impact of the pest was substantial with all leaves infested at the end of the last generation in all 3 years tested. The distribution of the leafminer in the canopy was homogeneous. The species is redescribed and illustrated, a lectotype is designated and a new synonymy is established.

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

Seasonal trends and the parasitoid complex of Chinese wax scale (Ceroplastes sinensis) was studied from July 2010 to February 2013. Six commercial citrus groves located in northeastern Spain were sampled fortnightly. Chinese wax scale completed a single annual generation. Egg oviposition started in May and continued until mid-July. Egg hatching began in mid-June, and in the first quarter of August, the maximum percentage of hatched eggs was reached. In the same groves, the parasitoid species of C. sinensis were determined together with their seasonal trends, relative abundance and occurrence on C. sinensis. Four hymenoptera were found parasitizing C. sinensis, mainly on third instars and females: Coccophagus ceroplastae (Aphelinidae), Metaphycus helvolus (Encyrtidae), Scutellista caerulea (Pteromalidae) and Aprostocetus ceroplastae (Eulophidae). The most abundant species was A. ceroplastae, corresponding to 54% of the parasitoids emerged. Coccophagus ceroplastae and M. helvolus represented 19%, whereas S. caerulea comprised 8% of the total. This study is the first published record of C. ceroplastae in Spain and the first record of M. helvolus on C. sinensis in Spain. Concerning the economical thresholds normally used, sampling plans developed for the management of C. sinensis in citrus groves should target population densities of around 12–20% of invaded twigs, equivalent to 0.2–0.5 females per twig. The sample size necessary to achieve the desired integrated pest management precision is 90–160 twigs per grove for the enumerative plan and about 160–245 twigs per grove for the binomial plan.

Granivory can play a pivotal role in influencing regeneration, colonization as well as abundance and distribution of plants. Due to their high abundance, nutrient content and longevity, seeds are an important food source for many animals. Among insects, carabid beetles consume substantial numbers of seeds and are thought to be responsible for a significant amount of seed loss. However, the processes that govern which seeds are eaten and are therefore prevented from entering the seedbank are poorly understood. Here, we assess if DNA-based diet analysis allows tracking the consumption of seeds by carabids. Adult individuals of Harpalus rufipes were fed with seeds of Taraxacum officinale and Lolium perenne allowing them to digest for up to 3 days. Regurgitates were tested for the DNA of ingested seeds at eight different time points post-feeding using general and species-specific plant primers. The detection of seed DNA decreased with digestion time for both seed species, albeit in a species-specific manner. Significant differences in overall DNA detection rates were found with the general plant primers but not with the species-specific primers. This can have implications for the interpretation of trophic data derived from next-generation sequencing, which is based on the application of general primers. Our findings demonstrate that seed predation by carabids can be tracked, molecularly, on a species-specific level, providing a new way to unravel the mechanisms underlying in-field diet choice in granivores.

Emerald ash borer (EAB), Agrilus planipennis, is a serious invasive forest pest in North America responsible for killing tens to hundreds of millions of ash trees since it was accidentally introduced in the 1990s. Although host-plant resistance and natural enemies are known to be important sources of mortality for EAB in Asia, less is known about the importance of different sources of mortality at recently colonized sites in the invaded range of EAB, and how these relate to host tree crown condition. To further our understanding of EAB population dynamics, we used a large-scale field experiment and life-table analyses to quantify the fates of EAB larvae and the relative importance of different biotic mortality factors at 12 recently colonized sites in Maryland. We found that the fates of larvae were highly dependent on EAB life stage and host tree crown condition. In relatively healthy trees (i.e., with a low EAB infestation) and for early instars, host tree resistance was the most important mortality factor. Conversely, in more unhealthy trees (i.e., with a moderate to high EAB infestation) and for later instars, parasitism and predation were the major sources of mortality. Life-table analyses also indicated how the lack of sufficient levels of host tree resistance and natural enemies contribute to rapid population growth of EAB at recently colonized sites. Our findings provide further evidence of the mechanisms by which EAB has been able to successfully establish and spread in North America.

Continuous ingestion of the phloem sap of plants by aphids can remove a significant amount of photoassimilates. Based on our earlier works, we hypothesized that due to the reduced aphid feeding time caused by antibiosis, wheat plants may achieve growth tolerance to aphids. We tested this hypothesis using three wheat cultivars, XY22 (Xiaoyan22), AK58 (Bainongaikang58) and XN979 (Xinong979) and the grain aphid, Sitobion avenae. In the choice test, S. avenae did not show any preference among the three wheat cultivars. However, S. avenae had a lower body weight and a lower intrinsic rate of increase when feeding on XY22 than on AK58 and XN979. The electrical penetration graph results indicated that S. avenae had significantly shorter mean and total phloem ingestion periods on XY22 than on AK58 or XN979. The aphids required a similar time to reach the phloem sap on the three wheat cultivars, but required more time to establish sustained phloem ingestion on XY22. These results suggest that the resistance factors of XY22 may be phloem based. Moreover, XY22 suffered less biomass loss in response to aphid infestation compared with XN979, suggesting that XY22 also had a better growth tolerance to S. avenae than XN979. Wheat resistance level to S. avenae was partially correlated with plant photosynthetic rates, and peroxidase activities. These results confirmed that the limitation in aphid feeding from plant phloem in wheat cultivar XY22 was related to antibiosis but not antixenosis, which caused XY22 tolerance to S. avenae.

Scaphoideus titanus is the invasive vector of the phytoplasma causing the Flavescence dorée in European vineyards. This epidemic is a serious threat to viticulture that has been increasing for more than 60 years in Europe. We studied the effect of synchrony with the plant phenology and the effect of plant-sap quality on the individual fitness. Thus, we conducted laboratory experiments to determine if insect hatchings were synchronized with grapevine bud break. We used two natural populations: one from a cold winter vineyard and one from a mild winter vineyard. In both cases, egg hatching was synchronized with bud break and leaf appearance. The phloem quality of the young and old leaves as a food source was analysed by high-performance liquid chromatography, and the effects on S. titanus growth were evaluated. Phloem composition varied with the grapevine cutting's age but also varied between leaves of different ages from the same plant. The older leaves were less nutritious because they had the highest carbon-to-nitrogen ratio and the lowest content of essential amino acids. Despite diverse phloem qualities, no fitness difference was observed. We found that the synchronization of egg hatchings with bud break is well regulated. However, the nymphs are not affected by the phloem-sap quality, suggesting that S. titanus may accept different food qualities and that egg hatching synchrony could contribute to population expansion in vineyards.

The Mediterranean corn borer, Sesamia nonagrioides, occurs sympatrically in the northeast of Spain with other lepidopteran pests such as Ostrinia nubilalis and Mythimna unipuncta. In this study, we evaluated the electrophysiological and behavioural response of mated and unmated females and males of S. nonagrioides to their own complete pheromone blend, to its own four components separately, and to the pheromone components of the sympatric species O. nubilalis and M. unipuncta. Results of the electroantennogram recordings revealed that females of S. nonagrioides can detect their own pheromone blend and its individual components. Moreover, our results show that unmated females and males of S. nonagrioides are more sensitive to the female pheromone, showing higher electrophysiological response than the mated females and males. Electroantennogram recordings showed that males and females can detect the major sexual pheromone component of O. nubilalis (Z)-11-tetradecenyl acetate and the minor component of the pheromone of M. unipuncta (Z)-9-hexadecenyl acetate. When the sex pheromone stimulus was presented in the dual-choice assays, gravid females of S. nonagrioides were attracted to both their own complete pheromone blend and one of their own minor pheromone components, (Z)-11-hexadecenal, but the major sexual pheromone component of O. nubilalis acts as a behavioural antagonist to the females.

The shoot beetles Tomicus minor, Tomicus yunnanensis, and Tomicus brevipilosus have been decimating Pinus yunnanensis trees for more than 30 years in Southwestern China. To understand the chemical ecological relationship between pines and Tomicus, and among the three beetle species, we compared the attraction of these beetles to damaged shoots, extracts from damaged shoots, and volatiles from damaged shoots collected by the dynamic headspace sampling method. Experiments were performed using a modified open-arena olfactometer. The male T. minor and both sexes of T. brevipilosus were more strongly attracted to damaged shoots than to undamaged shoots and they showed attraction to shoots damaged by the same species. Female T. minor and both sexes of T. yunnanensis were attracted to shoots damaged by female T. brevipilosus. The three beetle species were attracted to shoot extracts and dynamic headspace volatiles from shoots damaged by the same species and sex. Female T. minor and male T. yunnanensis were also attracted to dynamic headspace volatiles from shoots damaged by both sexes of T. brevipilosus. The results suggested that specific semiochemicals that are induced or produced by T. brevipilosus also attract T. minor and T. yunnanensis. The semiochemicals in damaged shoots affect the attraction of the three beetle species and play an important chemical communication role in weakening the host trees during the beetles’ shoot-feeding phase.

Xanthohumol, a prenylated flavonoid from hops, and a supercritical carbon dioxide extract of spent hops were studied for their antifeedant activity against stored product insect pests: Sitophilus granarius L., Tribolium confusum Duv. and Trogoderma granarium Everts. Xanthohumol exhibited medium deterrent activity against the adults of S. granarius L. and larvae of T. confusum Duv. The spent hops extract was more active than xanthohumol towards the adults of T. confusum Duv. The potential application of the crude spent hops extract as a feeding deterrent against the stored product pests is proposed.

The egg parasitoid Trissolcus vassilievi (Mayr) (Hymenoptera: Scelionidae) is a significant natural enemy of the sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae), the most important pest of wheat in Iran. This study examined the developmental time and egg-to-adult survival of two geographically separate populations of T. vassilievi on two corresponding host populations at five constant temperatures ranging from 15.0 to 35.0±1°C. No wasps of either population emerged at 15.0°C and the temperature threshold for development was similar between populations, ranging from 13.1±0.3 to 13.8±0.4°C for males and 12.2±0.1 to 12.6±0.1°C for females, but the thermal constant varied with gender and parasitoid population. Development of wasps from the colder Tabriz location was slower, with thermal constants for males and females of 172.6±3.1 and 204.1±1.2 degree-days, respectively, compared to Varamin wasps with 164.7±3.0 and 195.6±1.3 degree-days, respectively. Based on genetic inheritance patterns, reciprocal crosses between the two populations were expected to result in females with thermal phenotypes intermediate to their parental populations, and males that resembled their mothers. However, female progeny of crosses more closely resembled their maternal population, indicating a maternal effect on thermal phenotype. Furthermore, the magnitude of the maternal effect on the thermal constant was asymmetric and was more strongly expressed by Varmin than Tabriz females. These results suggest the possibility of using selective crosses between wasp populations, in combination with artificial selection in the laboratory, to tune the thermal phenotype of parasitoids to specific regions prior to augmentative releases.

Necrophoric behaviour is critical sanitation behaviour in social insects. However, little is known about the necrophoric responses of workers towards different developmental stages in a colony as well as its underlying mechanism. Here, we show that Solenopsis invicta workers display distinct necrophoric responses to corpses of workers and pupae. Corpses of workers killed by freezing (dead for <1 h) were carried to a refuse pile, but pupal corpses would take at least 1 day to elicit workers’ necrophoric response. Metarhizium anisopliae-infected pupal corpses accelerated the necrophoric behaviour of resident workers, with 47.5% of unaffected corpses and 73.8% infected corpses discarded by 1 day post-treatment). We found that fungus-infected pupal corpses had a higher concentration of fatty acids (palmitic acid, oleic acid and linoleic acid) on their surface. We experimentally confirmed that linoleic and oleic acids would elicit a necrophoric response in workers. The appearance of linoleic and oleic acids appeared to be chemical signals involved in recognition of pupal corpses, and M. anisopliae infection could promote the accumulation of fatty acids on surface of pupal corpses resulting in accelerated necrophoric responses of workers.

Obolodiplosis robiniae is native to North America and is an important introduced insect pest that forms leaf margin roll galls on species of genus Robinia (Fabaceae) in China. It was first detected in China in 2004, but subsequently spread and provoked local outbreaks. An analysis of a 676-bp sequence of the mitochondrial DNA cytochrome oxidase subunit I was conducted in 560 individuals from 28 populations, in order to (1) assess population genetic structuring and (2) explore possible explanations for the rapid spread and invasion success of O. robiniae. Yet, only four haplotypes were identified and the nucleotide diversity was low (π = 0.00005) and among the 560 specimens studied, only ten showed haplotypic variation involving no more than three substitutions. The result showed a low degree of genetic diversity among populations of the successful invasive gall midge, which suggested that the pest experienced a severe genetic bottleneck and a loss of genetic diversity after its introduction. The successful establishment and spread of O. robiniae in China is attributed to the wide distribution of its host plant, thus allowing ample opportunities for gene flow in the pest species, and to the advantageous life history characteristics of O. robiniae.

Quantification of the movement of insects at field and landscape levels helps us to understand their ecology and ecological functions. We conducted a meta-analysis on movement of carabid beetles (Coleoptera: Carabidae), to identify key factors affecting movement and population redistribution. We characterize the rate of redistribution using motility μ (L2 T−1), which is a measure for diffusion of a population in space and time that is consistent with ecological diffusion theory and which can be used for upscaling short-term data to longer time frames. Formulas are provided to calculate motility from literature data on movement distances. A field experiment was conducted to measure the redistribution of mass-released carabid, Pterostichus melanarius in a crop field, and derive motility by fitting a Fokker–Planck diffusion model using inverse modelling. Bias in estimates of motility from literature data is elucidated using the data from the field experiment as a case study. The meta-analysis showed that motility is 5.6 times as high in farmland as in woody habitat. Species associated with forested habitats had greater motility than species associated with open field habitats, both in arable land and woody habitat. The meta-analysis did not identify consistent differences in motility at the species level, or between clusters of larger and smaller beetles. The results presented here provide a basis for calculating time-varying distribution patterns of carabids in farmland and woody habitat. The formulas for calculating motility can be used for other taxa.

Population genetics of invading pests can be informative for understanding their ecology. In this study, we investigated population genetics of the invasive alfalfa weevil Hypera postica in Fukuoka Prefecture, Japan. We analyzed mitochondrial tRNALeu-COII, nuclear EF-1α gene fragments, and Wolbachia infection in relation to three leguminous host plants: Vicia angustifolia, Vicia villosa, and a new host Astragalus sinicus cultivated as a honey source and green manure crop. A parsimony network generated from mitochondrial gene sequences uncovered two major haplotypic groups, Western and Egyptian. In contrast to reported Wolbachia infection of the Western strain in the United States, none of our analyzed individuals were infected. The absence of Wolbachia may contribute to the stable coexistence of mitochondrial strains through inter-strain reproductive compatibility. Hypera postica genetic variants for the mitochondrial and nuclear genes were associated neither with host plant species nor with two geographic regions (Hisayama and Kama) within Fukuoka. Mitochondrial haplogroups were incongruent with nuclear genetic variants. Genetic diversity at the nuclear locus was the highest for the populations feeding on V. angustifolia. The nuclear data for A. sinicus-feeding populations indicated past sudden population growth and extended Bayesian skyline plot analysis based on the mitochondrial and nuclear data showed that the growth of A. sinicus-feeding population took place within the past 1000 years. These results suggest a shorter history of A. sinicus as a host plant compared with V. angustifolia and a recent rapid growth of H. postica population using the new host A. sinicus.