Small-bodied males invest in larger testes when highly ornamented.

Sperm competition and male mating rate are two non-mutually exclusive key evolutionary pressures selecting for larger testes within and across animal taxa. A few studies have tried to test the role of mating rate in the absence of sperm competition. Under the mating rate hypothesis, particular phenotypes of a given population that are expected to gain more mates (e.g., more ornamented males) are expected to make higher investments in testes size (a proxy for sperm production). We test this prediction in Polistes simillimus, a neotropical paper wasp in which females are single mated (no sperm competition) and males can mate with multiple partners. Testes size was predicted by body size (positive association), sexual ornamentation (negative association), and their interaction (among small males, testes size was positively related to ornamentation, but the opposite pattern was observed among large males). We propose that small-bodied well-ornamented males may face the highest risk of sperm depletion. Small-bodied males make relatively higher investment in testes size when highly ornamented. This strategy might be less profitable to large males, as they have overall larger testes. Our results provide strong evidence for the mating rate hypothesis.

The evolution of parasitism from mutualism in wasps pollinating the fig, Ficus microcarpa, in Yunnan Province, China.

Theory identifies factors that can undermine the evolutionary stability of mutualisms. However, theory's relevance to mutualism stability in nature is controversial. Detailed comparative studies of parasitic species that are embedded within otherwise mutualistic taxa (e.g., fig pollinator wasps) can identify factors that potentially promote or undermine mutualism stability. We describe results from behavioral, morphological, phylogenetic, and experimental studies of two functionally distinct, but closely related, Eupristina wasp species associated with the monoecious host fig, Ficus microcarpa, in Yunnan Province, China. One (Eupristina verticillata) is a competent pollinator exhibiting morphologies and behaviors consistent with observed seed production. The other (Eupristina sp.) lacks these traits, and dramatically reduces both female and male reproductive success of its host. Furthermore, observations and experiments indicate that individuals of this parasitic species exhibit greater relative fitness than the pollinators, in both indirect competition (individual wasps in separate fig inflorescences) and direct competition (wasps of both species within the same fig). Moreover, phylogenetic analyses suggest that these two Eupristina species are sister taxa. By the strictest definition, the nonpollinating species represents a "cheater" that has descended from a beneficial pollinating mutualist. In sharp contrast to all 15 existing studies of actively pollinated figs and their wasps, the local F. microcarpa exhibit no evidence for host sanctions that effectively reduce the relative fitness of wasps that do not pollinate. We suggest that the lack of sanctions in the local hosts promotes the loss of specialized morphologies and behaviors crucial for pollination and, thereby, the evolution of cheating.

A new species of Diolcogaster (Hymenoptera: Braconidae) from Brazil of potential use in the biological control of the pest Hypercompe brasiliensis (Stoll) (Lepidoptera: Erebidae).

Species of Diolcogaster parasitize Lepidoptera pests of commercial plants. The diversity of this genus is high, but few species of Diolcogaster have been described. The description of a new Diolcogaster species provides information for the biological control using this insect. This study presents the description and key notes on the biology of a new Diolcogaster parasitoid wasp. This species was reared from a caterpillar of Hypercompe brasiliensis collected after feeding on a Gloxinia perennis plant important to floriculture. Two complementary identification analyzes were performed on Diolcogaster adult bodies. The first was the analyses of its external morphology and the second its molecular analysis (mitochondrial DNA). The morphological analysis defined the insect as a new species of Diolcogaster, named Diolcogaster joanesi sp. nov. A maximum-likelihood (ML) analysis partially confirmed the morphological analysis, placing D. joanesi within a cluster including a previously identified species (Diolcogaster choi) and seven other morphospecies. The proximity of D. joanesi to D. choi is discussed and an updated key for all New World species of the xanthaspis group is provided. Twenty-eight adult wasps were obtained (22 females and six males) out of 50 cocoons which larvae emerged from the caterpillar host. The findings contribute to the broader understanding of Diolcogaster in the Neotropics and its potential for the biological control of lepidopteran defoliators.

Marimbondos: systematics, biogeography, and evolution of social behaviour of neotropical swarm-founding wasps (Hymenoptera: Vespidae: Epiponini).

Neotropical swarm-founding wasps are divided into 19 genera in the tribe Epiponini (Vespidae, Polistinae). They display extensive variation in several colony-level traits that make them an attractive model system for reconstructing the evolution of social phenotypes, including caste dimorphism and nest architecture. Epiponini has been upheld as a solid monophyletic group in most phylogenetic analyses carried out so far, supported by molecular, morphological and behavioural data. Recent molecular studies, however, propose different relationships among the genera of swarm-founding wasps. This study is based on the most comprehensive epiponine sampling so far and was analyzed by combining morphological, nesting and molecular data. The resulting phylogenetic hypothesis shows many of the traditional clades but still impacts the way certain behavioural characters, such as nest structure and castes, evolved, and thus requires some re-interpretations. Angiopolybia as sister to the remaining Epiponini implies that nest envelopes and a casteless system are plesiomorphic in the tribe. Molecular dating points to an early tribal diversification during the Eocene (c. 55-38 Ma), with the major differentiation of current genera concentrated in the Oligocene/Miocene boundary.

Wasp Waist and Flight: Convergent Evolution in Wasps Reveals a Link between Wings and Body Shapes.

Insect flight is made possible by different morphological structures: wings produce the lift, the thorax drives the wings' movements, and the abdomen serves as a secondary control device. As such, the covariation of these structures could reflect functional constraints related to flight performances. This study examines evolutionary convergences in wasp body shapes to provide the first evidence for morphological integration among insect wings, thorax, and abdomen. The shapes of the forewings and hind wings, thorax, and petiole (connecting abdomen and thorax) of 22 Vespidae species were analyzed using computerized tomography and geometric morphometrics. Results show a clear relationship between petiole and wings or thorax shapes but not between wings and thorax. Wasps with elongated bodies have pointed wings, both features thought to improve flight maneuverability. In contrast, stouter species have rounded wings, which may allow for higher flight speeds. These integration patterns suggest that multiple selective regimes on flight performance, some of them biased toward maneuverability or maximal speed, drove the morphological diversity in Vespidae. The results also suggest that wing shapes evolved under constraints related to the body type they have to lift. The abdomen morphology is thus another factor to take into account to understand the flight performance of insects.

Species- and size-related differences in dopamine-like immunoreactive clusters in the brain of Nasonia vitripennis and N. giraulti.

An extreme reduction in body size has been shown to negatively impact the memory retention level of the parasitic wasp Nasonia vitripennis. In addition, N. vitripennis and Nasonia giraulti, closely related parasitic wasps, differ markedly in the number of conditioning trials required to form long-term memory. These differences in memory dynamics may be associated with differences in the dopaminergic neurons in the Nasonia brains. Here, we used dopamine immunoreactivity to identify and count the number of cell bodies in dopaminergic clusters of normal- and small-sized N. vitripennis and normal-sized N. giraulti. We counted in total a maximum of approximately 160 dopaminergic neurons per brain. These neurons were present in 9 identifiable clusters (D1a, D1b, D2, D3, D4a, D4b, D5, D6 and D7). Our analysis revealed that N. giraulti had fewer cells in the D2 and D4a clusters but more in D4b, compared with normal-sized N. vitripennis. In addition, we found fewer cells in the D5 and D7 cluster of small-sized N. vitripennis compared to normal-sized N. vitripennis. A comparison of our findings with the literature on dopaminergic clusters in the fruit fly Drosophila melanogaster and the honey bee Apis mellifera indicates that clusters D2, D3 and D5 may play a role in memory formation in Nasonia wasps. The results from both the species comparison and the size comparison are therefore of high interest and importance for our understanding of the complex intricacies that underlie the memory dynamics of insects.

Quantitative genetics of wing morphology in the parasitoid wasp Nasonia vitripennis: hosts increase sibling similarity.

The central aim of evolutionary biology is to understand patterns of genetic variation between species and within populations. To quantify the genetic variation underlying intraspecific differences, estimating quantitative genetic parameters of traits is essential. In Pterygota, wing morphology is an important trait affecting flight ability. Moreover, gregarious parasitoids such as Nasonia vitripennis oviposit multiple eggs in the same host, and siblings thus share a common environment during their development. Here we estimate the genetic parameters of wing morphology in the outbred HVRx population of N. vitripennis, using a sire-dam model adapted to haplodiploids and disentangled additive genetic and host effects. The results show that the wing-size traits have low heritability (h2 ~ 0.1), while most wing-shape traits have roughly twice the heritability compared with wing-size traits. However, the estimates increased to h2 ~ 0.6 for wing-size traits when omitting the host effect from the statistical model, while no meaningful increases were observed for wing-shape traits. Overall, host effects contributed to ~50% of the variation in wing-size traits. This indicates that hosts have a large effect on wing-size traits, about fivefold more than genetics. Moreover, bivariate analyses were conducted to derive the genetic relationships among traits. Overall, we demonstrate the evolutionary potential for morphological traits in the N. vitripennis HVRx-outbred population, and report the host effects on wing morphology. Our findings can contribute to a further dissection of the genetics underlying wing morphology in N. vitripennis, with relevance for gregarious parasitoids and possibly other insects as well.

Mechanisms of ovipositor insertion and steering of a parasitic wasp.

Drilling into solid substrates with slender beam-like structures is a mechanical challenge, but is regularly done by female parasitic wasps. The wasp inserts her ovipositor into solid substrates to deposit eggs in hosts, and even seems capable of steering the ovipositor while drilling. The ovipositor generally consists of three longitudinally connected valves that can slide along each other. Alternative valve movements have been hypothesized to be involved in ovipositor damage avoidance and steering during drilling. However, none of the hypotheses have been tested in vivo. We used 3D and 2D motion analysis to quantify the probing behavior of the fruit-fly parasitoid Diachasmimorpha longicaudata (Braconidae) at the levels of the ovipositor and its individual valves. We show that the wasps can steer and curve their ovipositors in any direction relative to their body axis. In a soft substrate, the ovipositors can be inserted without reciprocal motion of the valves. In a stiff substrate, such motions were always observed. This is in agreement with the damage avoidance hypothesis of insertion, as they presumably limit the overall net pushing force. Steering can be achieved by varying the asymmetry of the distal part of the ovipositor by protracting one valve set with respect to the other. Tip asymmetry is enhanced by curving of ventral elements in the absence of an opposing force, possibly due to pretension. Our findings deepen the knowledge of the functioning and evolution of the ovipositor in hymenopterans and may help to improve man-made steerable probes.

Running in circles in phylomorphospace: host environment constrains morphological diversification in parasitic wasps.

Understanding phenotypic diversification and the conditions that spur morphological novelty or constraint is a major theme in evolutionary biology. Unequal morphological diversity between sister clades can result from either differences in the rate of morphological change or in the ability of clades to explore novel phenotype ranges. We combine an existing phylogenetic framework with new phylogenomic data and geometric morphometrics to explore the relative roles of rate versus mode of morphological evolution for a hyperdiverse group: cryptine ichneumonid wasps. Data from genomic ultraconserved elements confirm that cryptines are divided into two large clades: one specialized in the use of hosts that are deeply concealed under hard substrates, and another with a much more diversified host range. Using a phylomorphospace approach, we show that both clades have experienced similar rates of morphological evolution. Nonetheless, the more specialized group is much more restricted in morphospace occupation, indicating that it repeatedly evolved morphological change through the same morphospace regions. This is in agreement with our prediction that host use imposes constraints in the morphospace available to lineages, and reinforces an important distinction between evolutionary stasis as opposed to a scenario of continual morphological change restricted to a certain range of morphotypes.

No gains for bigger brains: Functional and neuroanatomical consequences of relative brain size in a parasitic wasp.

Heritable genetic variation in relative brain size can underlie the relationship between brain performance and the relative size of the brain. We used bidirectional artificial selection to study the consequences of genetic variation in relative brain size on brain morphology, cognition and longevity in Nasonia vitripennis parasitoid wasps. Our results show a robust change in relative brain size after 26 generations of selection and six generations of relaxation. Total average neuropil volume of the brain was 16% larger in wasps selected for relatively large brains than in wasps selected for relatively small brains, whereas the body length of the large-brained wasps was smaller. Furthermore, the relative volume of the antennal lobes was larger in wasps with relatively large brains. Relative brain size did not influence olfactory memory retention, whereas wasps that were selected for larger relative brain size had a shorter longevity, which was even further reduced after a learning experience. These effects of genetic variation on neuropil composition and memory retention are different from previously described effects of phenotypic plasticity in absolute brain size. In conclusion, having relatively large brains may be costly for N. vitripennis, whereas no cognitive benefits were recorded.

Brain structure differences between solitary and social wasp species are independent of body size allometry.

Evolutionary transitions in social behavior are often associated with changes in species' brain architecture. A recent comparative analysis showed that the structure of brains of wasps in the family Vespidae differed between solitary and social species: the mushroom bodies, a major integrative brain region, were larger relative to brain size in the solitary species. However, the earlier study did not account for body size effects, and species' relative mushroom body size increases with body size in social Vespidae. Here we extend the previous analysis by measuring the effects of body size variation on brain structure differences between social and solitary vespid wasps. We asked whether total brain volume was greater relative to body size in the solitary species, and whether relative mushroom body size was greater in solitary species, after accounting for body size effects. Both total brain volume and relative mushroom body volume were significantly greater in the solitary species after accounting for body size differences. Therefore, body size allometry did not explain the solitary versus social species differences in brain structure. The evolutionary transition from solitary to social behavior in Vespidae was accompanied by decreases in total brain size and in relative mushroom body size.

Large mothers produce progeny with high survival rates during the immature stage and large sizes at adulthood in a parasitoid species.

Parasitoid researchers have generally thought that the body size of the mother parasitoid does not affect the fitness performance of the progeny during the immature stage, as long as the progeny develop in the same environment. We reveal for the first time that this is not true for the parasitoid Echthrodelphax fairchildii (Hymenoptera: Dryinidae), which is parasitic on planthoppers. Large females ensured an increased survival rate for their progeny during the immature stage and a large body size at adult emergence. Maternal body size differentially affected the body sizes and survival rates of male and female progeny. Small females did not produce female progeny, and the survival rate of the female progeny increased more steeply with increasing maternal body size than that of the male progeny. Meanwhile, the body size of male progeny increased more steeply with increasing maternal body size. The influence of maternal body size on progeny survival to adult emergence has never been reported in insects before. In addition, large females were more likely to lay female eggs, suggesting that females control the sex ratio of progeny in response to their own body size.

Resolving the taxonomic status of biocontrol agents belonging to the Aphidius eadyi species group (Hymenoptera: Braconidae: Aphidiinae): an integrative approach.

Species that belong to the Aphidius eadyi group have been used as biocontrol agents against Acyrthosiphon pisum worldwide. However, despite their extensive use, there are still gaps in our knowledge about their taxonomy and distribution. In this study, we employed an integrative taxonomic approach by combining genetic analyses (mtDNA COI barcoding) with standard morphological analyses and geometric morphometrics of forewing shape. We identified three species within the A. eadyi species group, viz., A. smithi, A. eadyi and A. banksae. Genetic separation of all three species was confirmed, with mean genetic distances between species ranging from 5 to 7.4%. The following morphological characters were determined as the most important for separating species of the A. eadyi group: number and shape of costulae on the anterolateral part of the petiole, shape of the central areola on the propodeum, and shape and venation of the forewings. The differences in wing shape of all three species were statistically significant, but with some overlapping. We identified A. banksae as a widely distributed pea aphid parasitoid, whose known range covers most of the western Palaearctic (from the UK to Israel). Aphidius banksae is diagnosed and redescribed.

Ophelimus mediterraneus sp. n. (Hymenoptera, Eulophidae): a new Eucalyptus gall wasp in the Mediterranean region.

We report here for the first time the presence of Ophelimus mediterraneus sp. n. in Mediterranean Europe. This species appears to be closely related to Ophelimus maskelli, a well-known invasive pest of Eucalyptus. Based on molecular (cytochrome oxidase I, 28S), morphological (multivariate ratio analysis) and bio-ecological investigations, our study gives unambiguous relevant criteria that allow the discrimination between these two species. A full description of O. mediterraneus sp. n. is also provided. The geographic distribution of O. mediterraneus sp. n. as well as its impact on Eucalyptus species needs to be more widely assessed since its presence may have been confused with O. maskelli in their sympatric introduced areas. Further investigations of potential parasitoids in the native area may thus be welcomed to evaluate classical biological control achievability.

Impacts of seven insecticides on Cotesia flavipes (Cameron) (Hymenoptera: Braconidae).

The endoparasitoid wasp Cotesia flavipes (Cameron) (Hymenoptera: Braconidae) is inundatively released in Brazilian sugarcane plantations to control the sugarcane borers Diatraea saccharalis (Fabricius) and Diatraea flavipennella (Box) (Lepidoptera: Crambidae). In conjunction with these releases, several synthetic insecticides are used to control the neonate larvae of these pests. We assessed the lethal and transgenerational sublethal effects of seven of these insecticides on C. flavipes. Leaf discs were sprayed at the highest field concentrations of chlorantraniliprole, lambda-cyhalothrin + chlorantraniliprole, chlorfluazuron, triflumuron, lambda-cyhalothrin + thiamethoxam, tebufenozide, and novaluron. Distilled water was used as a negative control. Newly emerged females (24 h old) were placed in Petri dishes containing the treated leaves, and the lethal and transgenerational sublethal effects were assessed for the next two generations. Lambda-cyhalothrin + chlorantraniliprole and lambda-cyhalothrin + thiamethoxam caused 100% mortality of the parasitoid and were highly persistent, causing more than 30% mortality at 30 days after spraying. Chlorantraniliprole, chlorfluazuron, novaluron, and triflumuron did not cause significant mortality compared to the negative control, but did have transgenerational sublethal effects. The length of the tibia of the right posterior leg, used as a growth measurement, was reduced in the progeny (F1 generation) of exposed female parasitoids. In addition, chlorantraniliprole increased and chlorfluazuron reduced the proportion of females in the F1 generation, whereas novaluron reduced the proportion of females in the F2 generation. Overall, only tebufenozide was considered harmless to C. flavipes. The results of this study suggest that lambda-cyhalothrin + chlorantraniliprole and lambda-cyhalothrin + thiamethoxam are harmful to C. flavipes, although field studies are needed to obtain results for actual sugarcane crops.

The rare genus Leptofoenus Smith, 1862 (Hymenoptera: Pteromalidae, Leptofoeninae): new records, and aspects of its distribution.

Despite being the largest and most remarkable species of Chalcidoidea, species of Leptofoeninae are very rarely collected. Here, we expand the distribution range extension of the Leptofoenus howardi (Ashmead, 1895) for Rondônia and Minas Gerais, Leptofoenus stephanoides (Roman, 1920) for Rondônia, Espírito Santo and São Paulo and Leptofoenus westwoodi (Ashmead, 1895) for Roraima, Amazonas, Rondônia, Bahia, Minas Gerais, Rio de Janeiro and Rio Grande do Sul.

Structure and Sense Organs of Ovipositors of an Endoparasitoid Aprostocetus causalis and an Ectoparasitoid Quadrastichus mendeli in Leptocybe spp.

Little is known of the olfactory mechanisms of host detection in the ovipositors of endoparasitoids and ectoparasitoids. An endoparasitoid Aprostocetus causalis La Salle & Wu (Hymenoptera: Eulophidae) and an ectoparasitoid Quadrastichus mendeli Kim & La Salle (Hymenoptera: Eulophidae: Tetrastichinae) are the two parasitoids of the eucalyptus gall wasp Leptocybe spp. Structures and sense organs of ovipositors of A. causalis and Q. mendeli were studied using scanning and transmission electron microscopy, which provided essential information for exploring the mechanism of host detection by endoparasitoid and ectoparasitoid. The ovipositors of two parasitoids consisted of the first and second valvulae and ended in a pointed tip. There were three types of microtrichia, two types of sensilla chaetica, and one type of sensilla campaniformia on the ovipositors of A. causalis and Q. mendeli. However, Q. mendeli has the fourth type of microtrichia on the ovipositor. The morphology, types, distribution, length, and width of these sensilla and microtrichia were described, and their possible functions are discussed in conjunction with the stinging, oviposition, and the host selection process.

Genome-wide association analysis of host genotype and plastic wing morphological variation of an endoparasitoid wasp Asobara japonica (Hymenoptera: Braconidae).

Accumulating evidence suggests that genotype of host insects influences the development of koinobiont endoparasitoids. Although there are many potential genetic variations that lead to the internal body environmental variations of host insects, association between the host genotype and the parasitoid development has not been examined in a genome-wide manner. In the present study, we used highly inbred whole genome sequenced strains of Drosophila melanogaster to associate single nucleotide polymorphisms (SNPs) of host flies with morphological traits of Asobara japonica, a larval-pupal parasitoid wasp that infected those hosts. We quantified the outline shape of the forewings of A. japonica with two major principal components (PC1 and PC2) calculated from Fourier coefficients obtained from elliptic Fourier analysis. We also quantified wing size and estimated wasp survival. We then examined the association between the PC scores, wing size and 1,798,561 SNPs and  the association between the estimated wasp survival and 1,790,544 SNPs. As a result, we obtained 22, 24 and 14 SNPs for PC1, PC2 and wing size and four SNPs for the estimated survival with P values smaller than 10-5. Based on the location of the SNPs, 12, 17, 11 and five protein coding genes were identified as potential candidates for PC1, PC2, wing size and the estimated survival, respectively. Based on the function of the candidate genes, it is suggested that the host genetic variation associated with the cell growth and morphogenesis may influence the wasp's morphogenetic variation.

Testing the Dutilleul syndrome: host use drives the convergent evolution of multiple traits in parasitic wasps.

Common life-history aspects among independent lineages often result in the repeated evolution of suites of adaptive traits, or 'syndromes'. Such syndromes can be key avenues to understand relationships between morphological and ecological traits, but are rarely tested due to insufficient trait shift repetitions. We use a hyperdiverse lineage to investigate the evolution of a syndrome. Cryptine ichneumonid wasps that parasitize insects concealed in hard substrates display several traits that are putative adaptations to that end. Using a phylogenetic framework from a combined multigene molecular and morphological data set with 308 cryptine species, we tested whether these traits were part of a morphofunctional syndrome related to host use. Ancestral state estimations show multiple origins for six investigated traits, which are correlated to each other and to the use of deeply concealed hosts, suggesting adaptation. Putatively adaptive traits showed a much stronger link among themselves than with an assemblage of 49 other morphological traits. However, estimation of the order of evolution in adaptive traits showed no structured pattern. The results indicate that the challenge of attacking deeply concealed hosts induced the repeated evolution of a 'Dutilleul syndrome', named after the 'walker-through-walls' character from French literature. They also point towards a dynamic scenario in the evolution of complex functional systems. These findings highlight the power of morphology to illuminate poorly known aspects of natural history, and how hyperdiverse lineages can be used to understand the evolution of complex traits.

Caste-fate determination primarily occurs after adult emergence in a primitively eusocial paper wasp: significance of the photoperiod during the adult stage.

Independent-founding paper wasps constitute a major group of primitively eusocial insects, and when caste-fate determination occurs in temperate species of these wasps, particularly regarding whether it occurs before or after emergence, remains unclear. No critical morphological differences occur between potential queens of the next generation (often called gynes) and workers in primitively eusocial insects. The gynes of temperate species are characterized by diapausing, and the nutrients available during the larval stage have often been believed to determine caste fate. Short days usually induce diapause in temperate nonsocial insects, although few investigations of the effects of day length on caste-fate determination in paper wasps have been conducted. By exposing individuals to different combinations of short and long days during the immature and adult stages, we show for the first time that short days during the adult stage (but not during the immature stage) facilitated caste-fate determination toward gynes in a paper wasp. Moreover, the decision to diapause partly depended on changes in the photoperiod during the pupal and adult stages. The size of the adult also affected caste-fate determination, with diapause more likely to occur in large adults, but this size effect did not occur when individuals were exposed to many short days during the pupal stage. In addition, all adults except for a small proportion of smaller individuals prepared for diapause under short days. These findings suggest that the photoperiod is a higher priority cue than adult size.