Parasitic wasps are fascinating organisms that rely on a wide range of communication channels to locate their hosts. Associative learning for foraging kairomones has been demonstrated for various parasitic wasps, but little is known for parasitoids of Tephritidae flies. Psyttalia concolor (Hymenoptera: Braconidae) is a koinobiont parasitoid able to attack at least 14 tephritid pests. Females are innately attracted by some host-induced plant volatiles (HIPVs), whereas others of the same bouquet are unattractive. We hypothesize that females may detect unattractive HIPVs in association with key resources, such as food and hosts, learning to respond favourably to these cues in consecutive experiences. We evaluated associative learning for HIPVs in P. concolor females, testing if they are able to associate a food reward with the presence of different dosages of three HIPVs, thus developing a preference for an odour innately unattractive. Results demonstrated that P. concolor responded favourably to the learned cue in consecutive experiences. For all tested HIPVs (nonanoic acid, decanoic acid and geranyl acetone), regardless of dosage, trained females preferred the reward-associated odour, whereas naïve did not. Both HIPV-trained and naïve females did not show consistent differences in latencies when choosing HIPVs over blank. HIPV-trained and naïve wasps did not spend more time on HIPVs over blank. Odour learning is of adaptive importance for this generalist parasitoid, since it enhances host location efficiency by reducing the time wasted on the decision of where to search for hosts. From an applied perspective, these HIPVs could be used to train mass-reared P. concolor in pre-release, to potentially improve its efficacy in the field.

Specialization is an important attribute of a biological control agent. The maritime pine bast scale, Matsucoccus feytaudi Ducasse (Hemiptera Matsucoccidae), is an invasive species in Southeast France and the North of Italy. Iberorhyzobius rondensis Eizaguirre (Coleoptera: Coccinellidae), is a recently described ladybird species. Both adults and larvae are predaceous, feeding on egg masses of M. feytaudi, and are strongly attracted to M. feytaudi’s sex pheromone. To evaluate the potential of I. rondensis as a biocontrol agent of the scale, we studied its niche breadth and prey range with emphasis on pine forests and hemipterans as tested prey. In this study, I. rondensis was found to achieve complete development only when fed on M. feytaudi egg masses (92.9% survival) and an artificial prey: eggs of Ephestia kuehniella Zeller (27.6% survival). From the 2nd instar onwards, complete development could be achieved using other prey species, although larvae had significantly higher mortality and slower development. In choice tests, M. feytaudi was the preferred prey. Surveys of the ladybird populations in the Iberian Peninsula revealed that it was found exclusively on Pinus pinaster Aiton, the sole host of M. feytaudi. The unusual specialization of I. rondensis, among other predaceous ladybirds, makes it an appropriate candidate for classical biological control of M. feytaudi.

The North American and European maize pest Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) was used to assess whether conditions of the natal field, subsequent laboratory rearing, or genetic population origin affect phenotypic traits of fitness, activity, or morphometrics. Standardized laboratory bioassays with large sample sizes revealed that none of the 16 tested traits, except crawling behaviours, appeared consistently stable across all seven tested colonies. Environmental conditions in the natal field of the F0 generation affected trait averages of the subsequently reared F1 generation in laboratory in ca. 47% of cases, and trait variability in 67% of cases. This was apparent for fitness and morphometrics, but less obvious for activity traits. Early generation laboratory rearing affected trait averages in ca. 56% of cases: morphometrics changed; fecundity and egg survival increased from F1 to F2. Trait variability increased or decreased in 38% of cases. Laboratory rearing for over more than 190 generations affected the trait averages in 60% of cases, reflected by decreases in flight activity and increases in body size, weight, and fecundity to some extent. It had little effect on trait variability, especially so for morphometric variability. The genetic population origin affected average levels of 55% and variability of 63% of phenotypic traits. A comparison among D. v. virgifera studies might be difficult if they use different populations or laboratory colonies. It is advised to consider possible effects of original field conditions, laboratory rearing, and population genetics when planning comparative studies targeting fitness, activity, or morphometric questions regarding Diabrotica species.

Studying aquatic benthic macroinvertebrates (BMIs) in the field requires accurate taxonomic identification, which can be difficult and time consuming. Conventionally, head capsule morphology has been used to identify wild larvae of Chironomidae. However, due to the number of species and possible damage and/or deformity of their head capsules, another supporting approach for identification is needed. Here, we provide hemoglobin (Hb) protein in hemolymph of chironomids as a new biomarker that may help resolve some of the ambiguities and difficulties encountered during taxonomic identification. Chironomids collected from two locations in Maine and New Jersey, USA were identified to the genus level and in some cases to the species-level using head capsule and body morphologies. The head capsule for a particular individual was then associated with a corresponding Hb protein profile generated from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE). Distinct Hb profiles were observed from one group (Thienemannimyia) and four genera (Chironomus, Cricotopus, Dicrotendipes, and Glyptotendipes) of chironomids. Several species were polymorphic, having more than one Hb profile and/or having bands of the same size as those of other species. However, major bands and the combination of bands could distinguish individuals at the genus and sometimes species-level. Overall, this study showed that Hb profiles can be used in combination with head capsule morphology to identify wild chironomids.

Polar terrestrial invertebrates are suggested as being vulnerable to temperature change relative to lower latitude species, and hence possibly also to climate warming. Previous studies have shown Antarctic and Arctic Collembola and Acari to possess good heat tolerance and survive temperature exposures above 30 °C. To test this feature further, the heat tolerance and physiological plasticity of heat stress were explored in the Arctic collembolan, Megaphorura arctica, from Svalbard and the Antarctic midge, Eretmoptera murphyi, from Signy Island. The data obtained demonstrate considerable heat tolerance in both species, with upper lethal temperatures ≥35 °C (1 h exposures), and tolerance of exposure to 10 and 15 °C exceeding 56 days. This tolerance is far beyond that required in their current environment. Average microhabitat temperatures in August 2011 ranged between 5.1 and 8.1 °C, and rarely rose above 10 °C, in Ny-Ålesund, Svalbard. Summer soil microhabitat temperatures on Signy Island have previously been shown to range between 0 and 10 °C. There was also evidence to suggest that E. murphyi can recover from high-temperature exposure and that M. arctica is capable of rapid heat hardening. M. arctica and E. murphyi therefore have the physiological capacity to tolerate current environmental conditions, as well as future warming. If the features they express are characteristically more general, such polar terrestrial invertebrates will likely fare well under climate warming scenarios.

Traditional morphology-based taxonomy of eriophyoid mites (Acari: Eriophyoidea) has been challenged by molecular-based technologies in the detection of cryptic species. However, the implications of such cryptic diversity appear to differ when methods based on different types of data are used. Here, samples of a host-associated eriophyoid mite species, Tetra pinnatifidae, collected from different host plants and localities are evaluated. The congruence of results based on morphometric (32 characters), mitochondrial (16S), and nuclear (28S) data were evaluated and showed a host-associated cryptic diversity dividing this morphospecies into several groups/clades that were morphometrically indistinguishable. In comparison, the 16S data confirmed cryptic speciation and intra-clade host-associated diversity, while 28S did not. In contrast, 28S data revealed potential gene flow between host-associated populations. High mitochondrial divergence, as well as low nuclear and morphological divergence indicated very recent stage of cryptic diversity of this eriophyoid mite.

Area-wide sterile insect technique (SIT) programs assume that offspring reduction of the target population correlates with the mating success of the sterile males released. However, there is a lack of monitoring tools to prove the success of these programs in real-time. Field-cage tests were conducted under the environmental conditions of the Mediterranean coast of Spain to estimate: (a) the mating success of sterile Vienna-8 (V8) Ceratitis capitata males using molecular markers and (b) their efficacy to reduce C. capitata populations under six release ratios of wild females to wild males to V8 males (1:0:0, 1:1:0, 1:1:1, 1:1:5, 1:1:10, and 1:1:20). Statistical models were developed to predict: (a) the number of females captured in traps, (b) sperm ID (sterile or not) in spermathecae of the trapped females, and (c) the viable offspring produced, using release ratio and temperature as predictors. The number of females captured was affected by relative humidity. However, its influence in the model was low. Female captures were significantly higher in ratios 1:0:0 compared to ratios where V8 males were released. The proportion of V8 sperm in spermathecae increased with temperature and with the number of V8 males released, but leveled off between ratios 1:1:10 and 1:1:20. In all seasons, except winter (no offspring), viable offspring increased with temperature and was lowest for ratio 1:1:20. For the first time, a strong negative relationship between proportion of V8 sperm detected by molecular tools and C. capitata offspring was established. The models obtained should contribute to enhance the efficacy of SIT programs against this pest.

The egg parasitoid Telenomus podisi is a natural control agent of pentatomids, including Euschistus heros and Tibraca limbativentris, and success of parasitism is dependent upon the parasitoid finding the host. We tested the influence of host egg volatiles and the synthetic sex pheromone (zingiberenol) of T. limbativentris on chemotaxic behaviour of T. podisi, as well as, the impact of the original host on parasitoid selection. We used mated female T. podisi (48 h old) that emerged from the eggs of T. limbativentris or E. heros. The bioassays related to chemotaxy were performed in a Y-tube olfactometer and, to parasitism success, in laboratory and semi-field conditions. Telenomus podisi females that emerged from either the stink bug eggs, chose the pheromone more than control, or the pheromone plus eggs of E. heros in the semi-field bioassay, led to greater parasitism. Females that emerged from E. heros eggs chose egg volatiles from their original host rather than those from T. limbativentris, while females emerging from T. limbativentris, chose the egg volatiles of both hosts equally. When T. limbativentris was the original host, T. podisi females parasitized T. limbativentris over E. heros, while those emerging from E. heros exclusively parasitized E. heros eggs. These results demonstrated that T. podisi is more likely to parasitize the host in which it developed and that the original host can exert influence on the choice by those parasitoids. Understanding how the factors that mediate host–parasitoid communication are interrelated can help biological control programmes establish more effective and reliable tools with T. podisi.

The South American fruit fly, Anastrepha fraterculus, is a complex of cryptic species composed of at least seven morphotypes. Some of them, such as the Peruvian and Brazilian 1 morphotypes (which include Argentinean populations), exhibit strong pre-copulatory isolation, yet it is possible to obtain heterotypic crosses when forcing copulation of adults under laboratory conditions. The cross involving Peruvian males and Argentinean females produces F1 offspring with reduced viability in terms of egg hatch. This low hatchability could be caused by a reduced amount of sperm transferred to and stored by females mated with heterotypic males, which in turn could affect their post-copulatory behaviour. To test these hypotheses, we investigated sperm transfer and female mating and remating behaviour for homotypic and heterotypic crosses between adults of two morphotypes (Brazilian 1 [Argentina] and Peruvian [Peru]) of the A. fraterculus cryptic species complex. As reported before, Argentinean males and females mated earlier in the day than the other three mating combinations. Peruvian females engaged in shorter copulation times than Argentinean females. Peruvian females tended to store smaller quantities of sperm than Argentinean females, and almost a half of the crosses involving Argentinean males and Peruvian females were unsuccessful (no sperm transfer). However, there was no evidence that the cross between Peruvian males and Argentinean females resulted in storage of a critically small amount of sperm (posing risk of sperm shortage). Argentinean females were more willing to remate than Peruvian females, irrespective of male morphotype, but latency to remating was not affected by male or female morphotype. This study shows that mating behaviour differs between some of the A. fraterculus complex morphotypes, with female but not male morphotype determining female likelihood to remate.

In the present study predation parameters, i.e. consumption rate, conversion efficiency and growth rate, and total developmental duration of immature stages of two congeneric ladybirds, Coccinella septempunctata (L.) and Coccinella transversalis F., have been evaluated in presence of conspecific and heterospecific fourth instar larval and adult female tracks. We hypothesized that the semiochemicals within larval/adult female tracks might act as foraging/feeding deterrent pheromones (FDPs) and would reduce the predation parameters; and would prolong total developmental duration of ladybird predators. Results of the study positively affirmed our hypothesis. The deterrence in prey consumption and reduction in conversion efficiency and growth rate was density dependent with species-specific variations. Consumption rate, conversion efficiency, and growth rate of larval instars decreased and the total developmental duration of immature stages increased when exposed to an increasing density of zero, two, three, and four conspecific/heterospecific larval/adult female tracks. Between ladybird species, C. septempunctata had higher consumption rate, growth rate, and total developmental durations, whereas conversion efficiency was higher in C. transversalis. Despite the presence of semiochemical tracks as foraging barriers, early instars showed higher conversion efficiencies and growth rates, whereas fourth instars had highest consumption rate in all experimental treatments. The present study, therefore, suggests that semiochemical tracks significantly reduce the predation potential and prolong developmental duration of conspecifics and heterospecifics. They, thus behave as FDP.

In the Western Himalayas, Drosophila nepalensis is more abundant during the colder and drier winter than the warmer rainy season but the mechanistic bases of such adaptations are largely unknown. We tested effects of developmental plasticity on desiccation-related traits (body size, body melanization and water balance traits) that may be consistent with changes in seasonal abundance of this species. D. nepalensis grown at 15°C has shown twofold higher body size, greater melanization (∼15-fold), higher desiccation resistance (∼55 h), hemolymph as well as carbohydrate content (twofold higher) as compared with corresponding values at 25°C. Water loss before succumbing to death was much higher (∼16%) at 15°C than 25°C. Developmental plastic effects on body size are associated with changes in water balance-related traits (bulk water, hemolymph and dehydration tolerance). The role of body melanization was evident from the analysis of assorted darker and lighter flies (from a mass culture of D. nepalensis reared at 21°C) which lacked differences in dry mass but showed differences in desiccation survival hours and rate of water loss. For adult acclimation, we found a slight increase in desiccation resistance of flies reared at lower growth temperature, whereas in flies reared at 25°C such a response was lacking. In D. nepalensis, greater developmental plasticity is consistent with its contrasting levels of seasonal abundance. Finally, in the context of global climate change in the Western Himalayas, D. nepalensis seems vulnerable in the warmer season due to lower adult as well as developmental acclimation potential at higher growth temperature (25°C).

The mango blossom gall midge, Procontarinia mangiferae, is a multivoltine species that induces galls in inflorescences and leaves of the mango tree, Mangifera indica. In subtropical Reunion Island, populations of P. mangiferae are observed all-year round, but the pattern and the role of dormancy in their life cycle have never been documented. We performed field and laboratory experiments using more than 15,000 larvae. We demonstrated that a larval diapause may affect a part of the midge population, regardless of the season. The total duration of the diapause varied from 6 weeks to more than 1 year. One year of field monitoring showed that the highest incidence of diapause was observed in larvae collected during the summer from mango leaves, where it affected approximately one-third of the individuals. This facultative diapause allows the permanent presence of P. mangiferae in the orchards. By recording diapause duration during 22 weeks under controlled conditions, we showed that high temperatures (26 °C) increased diapause duration and extended the range of the dates of diapause emergence, whereas cool temperatures (20 °C) shortened diapause duration and shortened the range of the dates of emergence from diapause. A temperature decrease from 26 to 20 °C triggered the emergence of diapausing individuals. These mechanisms ensure the synchronization of the emergence of diapausing individuals with the appearance of mango inflorescences, which is also induced by cool winter temperatures.

Members of the family Cimicidae (Heteroptera: Cimicomorpha) are temporary bloodsuckers on birds and bats as primary hosts and humans as secondary hosts. Acanthocrios furnarii (2n=12=10+XY, male) and Psitticimex uritui (2n=31=28+X1X2Y, male) are two monotypic genera of the subfamily Haematosiphoninae, which have achiasmatic male meiosis of collochore type. Here, we examined chromatin organization and constitution of cimicid holokinetic chromosomes by determining the amount, composition and distribution of constitutive heterochromatin, and number and location of nucleolus organizer regions (NORs) in both species. Results showed that these two bloodsucker bugs possess high heterochromatin content and have an achiasmatic male meiosis, in which three regions can be differentiated in each autosomal bivalent: (i) terminal heterochromatic regions in repulsion; (ii) a central region, where the homologous chromosomes are located parallel but without contact between them; and (iii) small areas within the central region, where collochores are detected. Acanthocrios furnarii presented a single NOR on an autosomal pair, whereas P. uritui presented two NORs, one on an autosomal pair and the other on a sex chromosome. All NORs were found to be associated with CMA3 bright bands, indicating that the whole rDNA repeating unit is rich in G+C base pairs. Based on the variations in the diploid autosomal number, the presence of simple and multiple sex chromosome systems, and the number and location of 18S rDNA loci in the two Cimicidae species studied, we might infer that rDNA clusters and genome are highly dynamic among the representatives of this family.

Effective pest management with lower amounts of pesticides relies on accurate prediction of insect pest growth rates. Knowledge of the factors governing this trait and the resulting fitness of individuals is thus necessary to refine predictions and make suitable decisions in crop protection. The European grapevine moth, Lobesia botrana, the major pest of grapes in Europe, is responsible for huge economic losses. Larvae very rarely leave the grape bunch on which they were oviposited and thus cannot avoid intraspecific competition. In this study, we determined the impact of intraspecific competition during the larval stage on development and adult fitness in this species. This was tested by rearing different numbers of larvae on an artificial diet and measuring developmental and reproductive life history traits. We found that intraspecific competition during larval development has a slight impact on the fitness of L. botrana. The principal finding of this work is that larval density has little effect on the life history traits of survivors. Thus, the timing of eclosion, duration of subsequent oviposition, fecundity appears to be more uniform in L. botrana than in other species. The main effect of larval crowding was a strong increase of larval mortality at high densities whereas the probability of emergence, sex ratio, pupal mass, fecundity and longevity of mated females were not affected by larval crowding. Owing to increased larval mortality at high larval densities, we hypothesized that mortality of larvae at high densities provided better access to food for the survivors with the result that more food was available per capita and there were no effect on fitness of survivors. From our results, larval crowding alters the reproductive capacity of this pest less than expected but this single factor should now be tested in interaction with limited resources in the wild.

Conopomorpha sinensis Bradley (Lepidoptera: Gracilariidae) is one of the major fruit borer pests of litchi and longan in Southern China. Although chemical control is effective, alternative, biorational strategies are preferable, and should be developed. Predators play an important role in the biological control of agricultural pests, but an accurate method for the evaluation of predation in agriculture has not yet been developed. Here, we report a new, specific primer pair to amplify a C. sinensis cytochrome c oxidase subunit I (COI) sequence fragment that can be used to detect consumption of C. sinensis by its predators. C. sinensis DNA was found in several arthropods collected in the field, including the important C. sinensis predators Menochilus sexmaculata (Coccinellidae), Leucauge magnifica (Tetragnathidae), Propylea japonica (Coccinellidae), and Oxyopes sertatus (Oxyopidae). The detection rates of C. sinensis COI DNA in these predators were 39.3, 36.4, 27.3, and 27.2%, respectively. Laboratory consumption and hunting capacity analysis of M. sexmaculata and P. japonica adults indicated that they exhibit a Holling type II functional response on C. sinensis eggs under field temperatures. A polymerase chain reaction digestion analysis of M. sexmaculata and P. japonica adults after consumption of a single C. sinensis egg indicated that positive detection decreased with the extension of digestion time, and estimated prey DNA half-lives were 16.3 h in M. sexmaculata and 6.0 h in P. japonica. These data serve to characterize two major predators of C. sinensis with potential for biological control of C. sinensis in litchi orchards.

Temperature is a key environmental factor in determining the population size of Cnaphalocrocis medinalis in summer. High temperatures inhibit survival, development and fecundity of this insect. However, biological responses of female and male adults to heat shock, and physiological mechanism of high temperature suppressing population development are still ambiguous. We experimentally tested the impact of heat shock (5 h day−1) on biological traits, spermatogenesis and sperm transfer of adults of C. medinalis. The result showed that heat exposure to 39 and 40 °C for 5 h reduced longevity and copulation frequency of adults, and hatchability of eggs. Immediate survival rate of males was lower than that of females after 3 days of exposure to 41 °C. The oviposition period, copulation frequency, fecundity of adults and hatchability of eggs were significantly lower when male adults were exposed to 40 or 41 °C for 3 days. Heat shock decreased frequency and success rate of mating when males were exposed, and it also resulted in postponement of mating behaviour and prolongation of mating duration as both the female and male adults were exposed. Heat shock did not affect spermatogenesis, but significantly inhibited sperms maturation. Moreover, males could not ejaculate sperm into females during copulation when these male moths received heat shock. Heat shock remarkably suppressed mating behaviour and sperm transfer, which led to a dramatic decline of rice leaf folder populations.

Filariasis is a mosquito-borne disease that causes lymphedema and the main vector is Culex quinquefasciatus. A simple measure was taken to eradicate the vector using nanoemulsion. Eucalyptus oil nanoemulsion was formulated in various ratios comprising of eucalyptus oil, tween 80 and water by ultrasonication. The stability of nanoemulsion was observed over a period of time and 1:2 ratios of eucalyptus oil (6%) and surfactant (12%) was found to be stable. The formulated eucalyptus oil nanoemulsion was characterized by transmission electron microscopy and dynamic light scattering. The nanoemulsion droplets were found to have a Z-average diameter of 9.4 nm and were spherical in shape. The larvicidal activity of eucalyptus oil nanoemulsion and bulk emulsion was tested and compared. Our nanoemulsion showed higher activity when compared to bulk emulsion. The histopathology of larvae-treated and untreated nanoemulsion was analyzed. Furthermore, biochemical assays were carried out to examine the effect of nanoemulsion on biochemical characteristics of larvae. The treated larval homogenate showed decrease in total protein content and a significant reduction in the levels of acetylcholinesterase. The levels of acid and alkaline phosphatase also showed reduction as compared to control larval homogenate.

Infection of the commercially important silkworm, Bombyx mori by a tachnid parasitoid, Exorista bombycis induced activation of genes and cellular responses associated with apoptosis in integumental epithelial cells. Composite cellular profile showed initial autophagy, intermediate endoplasmic reticulum degranulation and deformed nucleus as well as later DNA fragmentation indicating apoptosis. Two cell death-associated proteins, autophagy 5-like (Atg5L) and apoptosis-inducing factor (AIF), in addition to caspase, are identified from the infected integumental epithelium through mass spectrometric analysis. Genes encoding these proteins showed age-dependent activation after the infection as revealed by quantitative expression analysis. Atg5 showed early upregulation in association with signs of autophagy whereas AIF showed late upregulation in association with DNA condensation and fragmentation. Expression of AIF showed negative correlation with that of Atg5 after the infection. On the other hand, in control, caspase expression showed positive correlation with AIF expression indicative of regulated expression in normal larval epithelium, which was absent after infection. Activation of Atg5, AIF and caspase genes in close association with different cell death events revealed the synchronized differential expression of apoptosis-associated genes in response to the macroparasitism. Enhanced expression of Atg5, AIF and caspase genes coupled with the appearance of cell death symptoms indicate parasitism-induced activation of genetic machinery to modulate cell death events in the epithelium, which was hither to unknown in invertebrate systems.