Small wasps, big muscles: Fore and hind leg modifications in chalcidoid wasps (Hymenoptera: Chalcidoidea).

A particularly conspicuous morphological feature in chalcidoid wasps are strikingly modified legs present in both males and females. It evolved convergently multiple times on either fore or hind legs implying strong evolutionary pressure and a prominent function in the wasps' life history. We investigate the external and internal morphology of the modified legs of five species of chalcidoid wasps representing four families (Ooderidae, Heydeniidae, Chalcididae, and Leucospidae), using light microscopy, scanning electron microscopy and micro computed tomography. We aimed to identify shared characteristics as well as differences between genera/species, leg pairs and sexes and to draw first conclusions about the shared or different functions. All species and sexes share the same general leg morphology, with enlarged femur, curved tibia and a huge flexor tibiae muscle. However, there are also genus/species-specific differences such as distinctive spine-like setae on the femur of Oodera spp., or leg pair-specific differences in the position of the extensor tibiae muscle. Shared characteristics imply a common primary function in which strong forces are required to pull the tibia against the femur while differences imply different secondary functions. Both primary and secondary functions have yet to be revealed beyond informed speculations.

Taxonomic revision of the Stenodynerus fastidiosissimus species-group in Western Europe and North Africa (Hymenoptera: Vespidae: Eumeninae).

The fastidiosissimus species-group of Stenodynerus de Saussure, 1863 is revised in Western Europe and North Africa, combining morphological data and DNA barcoding. Six species are recognized: S. difficilis (Morawitz, 1867) stat. resurr. (= S. fastidiosissimus auct.), S. fastidiosissimus (de Saussure, 1855), S. laborans (Costa, 1882) stat. resurr., S. montanus Selis, sp. nov., S. muelleri (Dusmet, 1917) (= Stenodynerus gusenleitneri Giordani Soika, 1986 syn. nov.), and S. rufescens Giordani Soika, 1977 stat. nov. Lectotypes are designated for Odynerus fastidiosissimus de Saussure, 1855 and Odynerus insularis Andr, 1883 non Smith, 1859. A key for the identification of members of this species-group is provided. DNA barcodes are published for every species, representing the first available sequences for the fastidiosissimus species-group.

Filling gaps in the knowledge of the Colombian Darwin wasps (Hymenoptera: Ichneumonidae): new records, description of unknown males, and taxonomical notes.

Darwin wasps (Hymenoptera: Ichneumonidae) are parasitoids and a natural enemy of a wide range of holometabolous insects and spiders and are considered one of the largest families in the animal kingdom. Unfortunately, there are great gaps in the knowledge of this family in Colombia. This study aims to expand the knowledge of Darwin wasps in Colombia, through new records for the country and especially the department of Antioquia. We present the diagnosis and digital images of the recorded species, and we describe for the first time the males of three species of Dolichomitus Smith, 1877: D. mariajosae Araujo & Pádua, 2020; D. menai Araujo & Pádua, 2020; D. orejuelai Araujo & Pádua, 2020.

First record of Vespa crabro Linnaeus (Hymenoptera: Vespidae) in western North America with a review of recorded species of Vespa Linnaeus in Canada.

Vespa crabro Linnaeus is newly reported as an adventive species in British Columbia, Canada which is the first record of this invasive species in western North America. The specimen of V. crabro was identified using morphological diagnostic keys and by comparison to authoritatively identified specimens. DNA barcoding provided support that the British Columbia specimen is conspecific with sequenced specimens of V. crabro. It is not possible to be certain of the origin of the specimen, but the DNA barcode was identical to sequence from specimens of V. crabro from South Korea. DNA barcoding was also performed on morphologically identified specimens of Vespa simillima and Vespa soror collected previously in British Columbia and the sequences were closest to V. simillima and V. soror Genbank sequences, respectively. There is no evidence that any of these species have established populations in the province. We provide diagnostic morphological characters to distinguish Canadian Vespa species from each other including Vespa mandarinia which has recently established populations in British Columbia and Washington State, USA. The potential detrimental impacts of each species are discussed.

Metamorphosis and denucleation of the brain in the miniature wasp Megaphragma viggianii (Hymenoptera: Trichogrammatidae).

Holometabolan brains undergo structural and allometric changes and complex reorganizations during metamorphosis. In minute egg parasitoids, brain formation is shifted to the late larva and young pupa, due to extreme de-embryonization. The brains of Megaphragma wasps undergo denucleation, the details of which remained unknown. We describe the morphological and volumetric changes in the brain of Megaphragma viggianii (Trichogrammatidae) during pupal development with emphasis on the lysis of nuclei and show that the absolute and relative volume of the brain decrease by a factor of 5 from prepupa to adult at the expense of the cell body rind. The first foci of lysis appear during early pupal development, but most nuclei (up to 97%) are lost between pharate adult and adult. The first signs of lysis (destruction of the nuclear envelopes) occur in pupae with red eyes. The number of lysis foci (organelle destruction and increasing number of lysosomes and degree of chromatin compaction) strongly increases in pupae with black eyes. The cell body rind volume strongly decreases during pupal development (in larger insects it increases slightly or remains unchanged). Elucidation of the lysis of nuclei in neurons and of the functioning of an anucleate brain is an important objective for neuroscience.

Cuticular hydrocarbons as caste-linked cues in Neotropical swarm-founding wasps.

Wasps (Vespidae) are important organisms to understand the evolution of social behaviour. Wasps show different levels of sociality, which includes solitary to highly eusocial organisms. In social insect species, queens and workers differ in physiology and morphology. The Neotropical swarm-founding wasps (Epiponini) show a variety of caste syndromes. In this clade, the caste-flexibility is a unique characteristic, in which workers can become queens and swarm to start a new nest. The investigation of the caste system comparing several Epiponini species show a clear-cut morphological distinction between queens and workers, with a morphological continuum between queens and workers. However, whether cuticular hydrocarbons (CHCs) are used as cues for caste recognition in swarm-founding wasps is still unknown. We studied whether CHCs may display caste-linked differences in eleven species of Epiponini wasps and if CHCs differences would follow morphological patterns. Our results suggest that queens and workers of Epiponini wasps are chemically different from each other at two levels, qualitatively and quantitatively, or merely quantitatively. This variation seems to exist regardless of their morphological traits and may be useful to help us understanding how chemical communication evolved differently in these species.

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.

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.

Re-collection and identity of Ooencyrtus californicus (Hymenoptera: Encyrtidae), and its new synonym, Ooencyrtus lucidus.

Girault (1917) very briefly described and diagnosed the encyrtid wasp species Ooencyrtus californicus Girault (Hymenoptera: Encyrtidae) with the following limited data (p. 22): "Sacramento, California, from bug eggs on Pinus sabiniana, September". The first author examined its two syntypes, poorly mounted on a slide, of which only parts of the four antennae and a slightly damaged fore wing remain (Triapitsyn et al. 2020). They concluded that this species was better considered a nomen dubium until fresh specimens could be collected from the same host plant in or near the type locality. In July 2019, an effort was made to re-collect O. californicus from foothill (or gray) pine, Pinus sabiniana (Pinaceae), in the Oakhurst area of Madera County, California, USA, where this pine is abundant, but no specimens were captured. At the same time, a very similar insect was collected from sentinel eggs of the invasive stink bug Bagrada hilaris (Burmeister) in Riverside, California. This insect was cultured in the laboratory as part of a B. hilaris biological control program. Due to the poor preservation of the type specimens and in the absence of genetic evidence, it was impossible to positively and properly attribute specimens of this newly collected parasitoid to O. californicus. Therefore, to provide a much needed scientific name for this native egg parasitoid, it was described as Ooencyrtus lucidus Triapitsyn Ganjisaffar in Triapitsyn et al. (2020). The two nominal species were separated based on minor differences in the proportions of the scape, pedicel, and first funicular segment of the female antenna (Triapitsyn et al. 2020). John S. Noyes, world expert on Encyrtidae and one of the reviewers of Triapitsyn et al. (2020), disagreed with the erection of O. lucidus as a new taxon based on the morphological similarities of the female antenna with that of O. californicus, so the description was published with the understanding that the two might be conspecific. But to settle this one way or another new specimens of O. californicus had to be collected to allow morphological and molecular analyses.

A New Species of Liosphex Townes, 1977 (Hymenoptera: Rhopalosomatidae) from the Amazon Forest and New Records for the Genus in Brazil.

Liosphex Townes, 1977 is a genus of solitary wasps belonging to the family Rhopalosomatidae, whose larvae develop as ectoparasitoids of crickets. In this paper, both sexes of a new species of the genus are described, Liosphex amazonensis Bulbol & Somavilla sp. n. Three other species, Liosphex achuar Lohrmann, 2010, Liosphex atratus Lohrmann, 2010, and Liosphex bribri Lohrmann, 2010, are recorded for the first time in Brazil. The male of Liosphex guanabara Lohrmann, 2010 is described for the first time, and the species is registered for the state of Bahia. In addition, the first detailed descriptions and illustrations of male genitalia of three species of the genus, i.e., L. amazonensis sp. n., L. atratus, and L. guanabara, are provided.

An Integrative Taxonomy of a New Species of Trichogramma Westwood (Hymenoptera: Trichogrammatidae) with High Reproductive Capacity.

A new species of Trichogramma Westwood (Hymenoptera: Trichogrammatidae) is described: Trichogramma foersteri sp. nov. Takahashi from eggs of Anticarsia gemmatalis Hübner, (Lepidoptera: Erebidae) a major soybean defoliating pest. The parasitoid was collected in São José dos Pinhais, Paraná State, Brazil, and molecular and morphological characters were used to confirm the identity of the new species. Preliminary biological data are provided to demonstrate its high capacity of parasitism. The new species is a potential candidate as a biological control agent against some lepidopteran-pests.

Morphology of sting apparatus of Chrysidoidea (Hymenoptera, Aculeata).

The main synapomorphy for Aculeata is the sting apparatus, which allows the female to envenom potential prey or hosts. The sting is the modified ovipositor which is not used for laying eggs anymore. Here, we explore the morphology of the sting apparatus within the families of Chrysidoidea. 27 skeletal structures were recognized, including three (dp1vf, dorsal projection of first valvifer; ppa, projection of posterior area of the second valvifer; vl9, ventral lap of tergite 9) that have not been observed previously, and 13 pairs of muscles, including four (superior dorsal T9-2vf (M5); inferior dorsal T9-2vf (M6); postero-lateral T9-2vf/mbr (M9); anterolateral 2vf/bl-2vv/fu (M11)) that have not been observed previously. Very conserved morphological patterns were observed; character support in the sting apparatus was found at the subfamily level, and within three families at the genus level. In addition, we describe the variation within the sting apparatus structures and musculature, propose evolutionary hypotheses about the function and evolution of the structures, and summarize phylogenetic conclusions for Chrysidoidea.

A New Genus and Two New Species of the Subfamily Nesomesochorinae Ashmead (Insecta: Hymenoptera: Ichneumonidae).

Nesomesochorinae is a rather small subfamily of Darwin wasps (Insecta: Hymenoptera: Ichneumonidae), only with three described genera. We recognize a fourth nesomesochorine genus from Perú and describe it as a new genus, Bina Shimizu & Alvarado gen. nov. The new genus is distinguished from previously described genera of Nesomesochorinae by the following combination of character states: mandible parallel-sided proximally and abruptly strongly narrowed ventroapically so that its lower apical tooth is much smaller and sharper than upper tooth; propodeum with lateromedian longitudinal carinae between anterior and posterior transverse carinae developed on more than posterior 0.9 so that the area superomedia is more or less enclosed; thyridium of 2nd metasomal tergite sometimes very large; 1st metasomal tergite stout and arched; body highly shiny. Bina gen. nov. comprises two species also described as new to science: B. huayrurae Shimizu & Alvarado sp. nov. (type species of Bina gen. nov.) and B. nigra Shimizu & Alvarado sp. nov. Identification keys to genera of Nesomesochorinae and to species of Bina gen. nov. are provided.

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.

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.

Taxonomic Status and Population Oscillations of Aphidius colemani Species Group (Hymenoptera: Braconidae) in Southern Brazil.

Aphidius colemani (Viereck) was reported in Brazil before the Biological Control Program of Wheat Aphids (BCPWA) when Mediterranean genotypes were introduced from France and Israel. This species was re-described as a complex called A. colemani group composed of three species. Consequently, uncertainty remains about which parasitoid of the group is occurring in southern Brazil. This study has two main objectives: (i) re-examine the species status of A. colemani group collected during the introduction of parasitoids and from a 10-year (2009-2018) monitoring program in wheat fields in northern Rio Grande do Sul (RS), Brazil; (ii) describe the variation in the population density of parasitoids and its association with meteorological factors during this period. We examined 116 specimens from the Embrapa Wheat entomological collection, and those collected in Moericke traps in Coxilha, RS. All the parasitoids of the A. colemani group from the BCPWA period were identified as Aphidius platensis (Brèthes). In traps, 6541 cereal aphid parasitoids were collected, of which 61.9% (n = 4047) were from A. colemani group and all those were identified as A. platensis. Temperature was the factor that effected population density with the highest number of parasitoids recorded in the winter months. Sex ratio changed between years varying from 0.50 to 0.97. The parasitoid A. platensis was the only species in the A. colemani group sampled during 10 years of monitoring.

A partial genome assembly of the miniature parasitoid wasp, Megaphragma amalphitanum.

Body size reduction, also known as miniaturization, is an important evolutionary process that affects a number of physiological and phenotypic traits and helps animals conquer new ecological niches. However, this process is poorly understood at the molecular level. Here, we report genomic and transcriptomic features of arguably the smallest known insect-the parasitoid wasp, Megaphragma amalphitanum (Hymenoptera: Trichogrammatidae). In contrast to expectations, we find that the genome and transcriptome sizes of this parasitoid wasp are comparable to other members of the Chalcidoidea superfamily. Moreover, compared to other chalcid wasps the gene content of M. amalphitanum is remarkably conserved. Intriguingly, we observed significant changes in M. amalphitanum transposable element dynamics over time, in which an initial burst was followed by suppression of activity, possibly due to a recent reinforcement of the genome defense machinery. Overall, while the M. amalphitanum genomic data reveal certain features that may be linked to the unusual biological properties of this organism, miniaturization is not associated with a large decrease in genome complexity.

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.

Direct evidence for eudicot pollen-feeding in a Cretaceous stinging wasp (Angiospermae; Hymenoptera, Aculeata) preserved in Burmese amber.

Angiosperms and their insect pollinators form a foundational symbiosis, evidence for which from the Cretaceous is mostly indirect, based on fossils of insect taxa that today are anthophilous, and of fossil insects and flowers that have apparent anthophilous and entomophilous specializations, respectively. We present exceptional direct evidence preserved in mid-Cretaceous Burmese amber, 100 mya, for feeding on pollen in the eudicot genus Tricolporoidites by a basal new aculeate wasp, Prosphex anthophilos, gen. et sp. nov., in the lineage that contains the ants, bees, and other stinging wasps. Plume of hundreds of pollen grains wafts from its mouth and an apparent pollen mass was detected by micro-CT in the buccal cavity: clear evidence that the wasp was foraging on the pollen. Eudicots today comprise nearly three-quarters of all angiosperm species. Prosphex feeding on Tricolporoidites supports the hypothesis that relatively small, generalized insect anthophiles were important pollinators of early angiosperms.