Many hemipteroids are major pests and vectors of microbial pathogens, infecting crops. Saliva of the hemipteroids is critical in enabling them to be voracious feeders on plants, including the economically important ones. A plethora of hemipteroid salivary enzymes is known to inflict stress in plants, either by degrading the plant tissue or by affecting their normal metabolism. Hemipteroids utilize one of the following three strategies of feeding behaviour: salivary sheath feeding, osmotic-pump feeding and cell-rupture feeding. The last strategy also includes several different tactics such as lacerate-and-flush, lacerate-and-sip and macerate-and-flush. Understanding hemipteroid feeding mechanisms is critical, since feeding behaviour directs salivary composition. Saliva of the Heteroptera that are specialized as fruit and seed feeders, includes cell-degrading enzymes, auchenorrhynchan salivary composition also predominantly consists of cell-degrading enzymes such as amylase and protease, whereas that of the Sternorhyncha includes a variety of allelochemical-detoxifying enzymes. Little is known about the salivary composition of the Thysanoptera. Cell-degrading proteins such as amylase, pectinase, cellulase and pectinesterase enable stylet entry into the plant tissue. In contrast, enzymes such as glutathione peroxidase, laccase and trehalase detoxify plant chemicals, enabling the circumvention of plant-defence mechanisms. Salivary enzymes such as M1-zinc metalloprotease and CLIP-domain serine protease as in Acyrthosiphon pisum (Aphididae), and non-enzymatic proteins such as apolipophorin, ficolin-3-like protein and ‘lava-lamp’ protein as in Diuraphis noxia (Aphididae) have the capacity to alter host-plant-defence mechanisms. A majority of the hemipteroids feed on phloem, hence Ca++-binding proteins such as C002 protein, calreticulin-like isoform 1 and calmodulin (critical for preventing sieve-plate occlusion) are increasingly being recognized in hemipteroid–plant interactions. Determination of a staggering variety of proteins shows the complexity of hemipteroid saliva: effector proteins localized in hemipteran saliva suggest a similarity to the physiology of pathogen–plant interactions.

Mutualistic relationships between honeydew-producing insects and ants have been widely recognized for several decades. Iridomyrmex rufoniger (Lowne) is the commonest ant species associated with black scale, Saissetia oleae (Olivier), in the citrus orchards of the mid latitudes of coastal New South Wales. Citrus trees with high densities of both red and black scale and high ant activity were identified and the results of excluding ants from half of those trees (using a polybutene band on each trunk) were compared with the results of not excluding ants from the other half. Trees with a low incidence of black scale and ants were also studied. Exclusion of ants from trees was soon followed by collapse of black scale populations because most individuals were asphyxiated by their own honeydew. Also, parasitism of the red scale by Encarsia perniciosi (Tower) and Encarsia citrina Craw was significantly higher than in the control trees over the following year, as was the predation rate on red scale due to three coccinellid predators, Halmus chalybeus (Boisduval), Rhyzobius hirtellus Crotch and Rhyzobius lophanthae (Blaisdell). In contrast, another coccinellid, Orcus australasiae (Boisduval), and a noctuid moth larva, Mataeomera dubia Butler, were seen in low numbers on banded (ant exclusion) trees, probably because of the low availability of their black scale prey, but were significantly higher on control trees apparently because of their invulnerability to ants.

In biological control, successful establishment of a natural enemy species depends on its adaptability in the introduced range including its ability to re-establish desired ecological interactions with the pest. These are affected by genetic parameters hitherto largely unresolved in biological control. The larger grain borer (LGB), Prostephanus truncatus, an invasive species from meso-America, is the most important post-harvest pest of maize in Africa. We studied the genetic structure of Teretrius nigrescens, a predatory beetle previously released for the control of the pest in Africa, to test the hypothesis that establishment patterns were a result of ecotype–environment mismatch and to follow up on our earlier reports of distinct lineages of the predator. We studied 13 populations of T. nigrescens, using 16 polymorphic microsatellite markers. Five genetic populations with a hierarchical structure and significant isolation by distance were detected. The most diverse population was found in southern Mexico, consistent with earlier lineage coexistence observations. Populations introduced to Africa maintained genetic similarity to local geographic populations of their area of origin. The more successful Benin releases were also more genetically diverse. Loss of rare alleles and a higher frequency of existing private alleles in some populations indicated population expansions following bottleneck events. Sustainable biological control should accommodate pest and natural enemy species, and monitor genetic changes associated with introduction and release.

The current study reports mortality and effects on cellular immune response of several entomopathogenic fungi including isoleates BB1, BB2 and BB3 of Beauveria bassiana, Metarhizium anisopliae, Isaria fumosoroseus and Lecanicilium lecanii against larvae of Chilo suppressalis. Prohemocytes, granulocytes, plasmatocytes and oenocytoids were identified as the main circulating hemocytes in the hemolymph of larvae using Giemsa staining solution. Entomopathogenic fungi caused differential mortality on larvae: BB1, BB3, M. anisopliae lead to the highest mortality on larvae and L. lecanii caused the lowest mortality. The highest numbers of total hemocytes were observed 3 h post-injection of B. bassiana isolates and 6 h for the other treatments. The highest numbers of plasmatocytes were observed 3 h post-injection of BB1 and Tween 80, whereas BB2, BB3, M. anisopliae, I.fumosoroseus and L. lecani caused plasmatocyte increase 6 h post-injection. Similar results were obtained in case of granulocytes but only Tween 80 showed the highest number of hemocytes 3 h post-injection. The highest numbers of nodules were found at various time intervals after injection of fungal isolates and latex bead. The highest activities of phenoloxidase were observed 12 h post-injection by BbB1, BbB3, M. anisopliae and latex bead; 3–6 h post-injection by BbB2, 6 h post-injection by I. fumosoroseus and 3–6 h post-injection by L. lecanii. Our data demonstrate the possibility of utilizing different fungal extracts in the field to help reduce the risk of resistance evolution in C. suppressalis and encourage experimentations aimed to increase the number of biological control agent for insect pests such as the striped rice stem borer C. suppressalis.

The mutualism between fig trees and their wasp pollinators is a model system for many ecological and evolutionary studies. However, the immature stages of pollinating fig wasps have rarely been studied. We monitored developing fig wasps of known ages and performed a series of dissections at 24 h intervals to identify key developmental traits of Ceratosolen solmsi marchali Mayr (Hymenoptera: Agaonidae), a pollinator of Ficus hispida L. (Moraceae). We identified where in the Ficus ovary eggs were deposited and time to hatch. We were also able to identify the timing and key underlying characters of five larval instars, three sub-pupal stages, and a single prepupal stage. We provide detailed morphological descriptions for the key stages and report some behavioral observations of the wasps in the several developmental stages we recorded. Scanning electron microscope images were taken.

Fruit flies (Diptera: Tephritidae) are major pests worldwide. The sterile insect technique, where millions of flies are reared, sterilized by irradiation and then released, is one of the most successful and ecologically friendly methods of controlling populations of these pests. The mating behaviour of irradiated and non-irradiated flies has been compared in earlier studies, but there has been little attention paid to the anti-predator behaviour of mass-reared flies, especially with respect to wild flies. Tephritid flies perform a supination display to their jumping spider predators in order to deter attacks. In this study, we evaluated the possibility of using this display to determine the anti-predator capabilities of mass-reared irradiated, non-irradiated flies, and wild flies. We used an arena setup and observed bouts between jumping spiders (Phidippus audax Hentz) and male Mexican fruit flies (Anastrepha ludens Loew). We show that although all flies performed a supination display to their predator, wild flies were more likely to perform a display and were significantly more successful in avoiding attack than mass-reared flies. We suggest that this interaction can be used to develop a rapid realistic method of quality control in evaluating anti-predator abilities of mass-reared fruit flies.

The seasonal dynamics of neutral genetic diversity and the insecticide resistance mechanisms of insect pests at the farm scale are still poorly documented. Here this was addressed in the green peach aphid Myzus persicae (Sulzer) (Hemiptera: Aphididae) in Central Chile. Samples were collected from an insecticide sprayed peach (Prunus persica L.) orchard (primary host), and a sweet-pepper (Capsicum annum var. grossum L.) field (secondary host). In addition, aphids from weeds (secondary hosts) growing among these crops were also sampled. Many unique multilocus genotypes were found on peach trees, while secondary hosts were colonized mostly by the six most common genotypes, which were predominantly sensitive to insecticides. In both fields, a small but significant genetic differentiation was found between aphids on the crops vs. their weeds. Within-season comparisons showed genetic differentiation between early and late season samples from peach, as well as for weeds in the peach orchard. The knock-down resistance (kdr) mutation was detected mostly in the heterozygote state, often associated with modified acetylcholinesterase throughout the season for both crops. This mutation was found in high frequency, mainly in the peach orchard. The super-kdr mutation was found in very low frequencies in both crops. This study provides farm-scale evidence that the aphid M. persicae can be composed of slightly different genetic groups between contiguous populations of primary and secondary hosts exhibiting different dynamics of insecticide resistance through the growing season.

Individual wild-caught sandflies from Iran were examined for infections of Wolbachia pipientis by targeting the major surface protein gene wsp of this intracellular α-proteobacterium. In total, 638 male and female sandflies were screened, of which 241 were found to be positive for one of three wsp haplotypes. Regardless of geographical origins and habitats, Phlebotomus (Phlebotomus) papatasi and other sandfly species were found to be infected with one common, widespread strain of A-group W. pipientis (Turk 54, GenBank accession EU780683; AY288297). In addition, a new A-group haplotype (Turk07, GenBank accession KC576916) was isolated from Phlebotomus (Paraphlebotomus) mongolensis and Phlebotomus (Pa.) caucasicus, and a new B-group haplotype (AZ2331, GenBank accession JX488735) was isolated from Phlebotomus (Larroussius) perfiliewi. Therefore, Wolbachia was found to occur in at least three of the incriminated vectors of zoonotic cutaneous leishmaniasis and zoonotic visceral leishmaniasis in different geographical regions of Iran. It may provide a new tool for the future control of leishmaniasis.

The tachinid fly Exorista larvarum (L.) (Diptera: Tachinidae) is a polyphagous larval endoparasitoid that deposits its eggs on the host exoskeleton of lepidopteran and tenthredinid larvae. The attachment of larval E. larvarum and the formation of the respiratory funnel were studied during infestation in the last larval instar of the wax moth, Galleria mellonella (L.) (Lepidoptera: Pyralidae). The tachinid larvae burrow through the host integument after hatching, using their robust cephalopharyngeal skeleton, leaving a dark spot at the point of their penetration as a result of host cuticle melanization. Endoparasitoid penetration induces the host cellular defence, resulting in the formation of a haemocyte capsule consisting of multi-cellular sheaths. This enveloping capsule later undergoes melanization, which is mostly obvious towards the posterior part of the endoparasitoid. The endoparasitoid uses the host encapsulation response to build a respiratory funnel from the modified host integument, leading to the host surface. The encapsulated larva remains attached to the respiratory funnel via an anal hook and cuticular spines until fully developed. Additional immunohistochemical analyses were used to study host–parasitoid interactions. Indirect immunofluorescence showed no labelling of potential tachinid antigens and confirmed no effect on the surrounding host tissues. A simulated parasitization with coated polybead microspheres revealed the mortal impact of tachinid antigens to the host. Hosts injected with antigen-coated polybeads died as a consequence of an acute and extensive immunological response to the tachinid antigens and not due to the trauma caused by foreign objects inside their body.

Mealybugs (Hemiptera: Pseudococcidae) are important pests for crops worldwide. Different species, cryptic taxa under the same species name or even populations within a species can differ in biological characteristics, such as phenology, resistance to insecticides, virus transmission and susceptibility to natural enemies. Therefore, their management efficacy depends on their accurate identification. Microsatellite genetic markers are efficient in revealing the fine-scale taxonomic status of insects, both at inter- and intra-specific level. Despite their potential uses, microsatellites have been developed only for one mealybug species so far. Hence, it is unclear whether microsatellites may be useful to assess mealybug population differentiation and structuring. In this work, we tested the feasibility of developing microsatellite markers in mealybugs by: (i) producing and characterizing microsatellite DNA libraries for three species: Pseudococcus viburni, Pseudococcus comstocki and Heliococcus bohemicus, and (ii) by developing and testing markers for Ps. viburni. The obtained libraries contained balanced percentages of dinucleotide (ranging from 15 to 25%) and trinucleotide (from 5 to 17%) motifs. The marker setup for Ps. viburni was successful, although 70% of the primers initially tested were discarded for a lack of polymorphism. Finally, 25 markers were combined in two multiplex polymerase chain reactions with 21 displaying no evidence of deviation from Hardy–Weinberg equilibrium. Ps. viburni markers were tested on one population from France and one from Chile. The markers revealed a significant genetic differentiation between the two populations with an Fst estimate of 0.266.

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.

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.

The sterile insect technique (SIT) is a non-chemical approach used to control major pests from several insect families, including Tephritidae, and entails the mass-release of sterile insects that reduce fertility of wild populations. For SIT to succeed, released sterile males must mature and compete with wild males to mate with wild females. To reach sexual maturity, the Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), must obtain adequate nutrition after adult emergence; however, in current SIT programs sterile B. tryoni receive a pre-release diet that lacks key nutrients required to sustain sexual development. The chief objective of this study was to determine whether pre-release yeast hydrolysate (YH) supplements affect the persistence and abundance of sexually mature sterile male B. tryoni under field conditions. Experiments were run in outdoor cages under conditions of low and high environmental stress that differed markedly in temperature and humidity, and in the field. Under low environmental stress conditions, survival of sterile B. tryoni was monitored in cages under three diet treatments: (i) sugar only, (ii) sugar plus YH or (iii) sugar plus YH for 48 h and sugar only thereafter. Under high environmental stress conditions survival of sterile B. tryoni was monitored in cages under four diet treatments: (i) white sugar only, (ii) brown sugar only, (iii) white sugar plus YH and (iv) brown sugar plus YH. In a replicated field study, we released colour-marked sterile B. tryoni from two diet regimes, YH-supplemented or YH-deprived, and monitored abundance of sexually mature males. In the low-stress cage study, there was no effect of diet, although overall females lived longer than males. In the high stress cage study, mortality was lower for YH-fed flies than YH-deprived flies and females lived longer than males. In the field, YH supplementation resulted in higher abundance of sexually mature sterile males, with 1.2 YH-fed flies trapped for every YH-deprived fly trapped. Under field conditions, YH supplementation can increase over-flooding ratios and hence may improve the effectiveness of SIT programmes.