The use of Proteinase K to access genitalia morphology, vouchering and DNA extraction in minute wasps.

Genitalia are rich source of characters in insect taxonomy. Usually, they are examined after dissection and cleaning with potassium hydroxide (KOH), procedure that may damage both genital morphological structures and intracellular molecular contents. Enzymatic procedure with Proteinase K has been used to clean muscle off the genitalia while extract DNA, but its damage to the genital structures has not been evaluated. Herein, we qualitatively compare the use of KOH and Proteinase K to prepare genital structures in minute insects (Hymenoptera: Bethylidae). We show that Proteinase K is better to preserve the genital structure and provides quality DNA.

Revision of Trichiscus benoît, 1956 (Hymenoptera, Bethylidae).

The species of the Afrotropical genus Trichiscus are revised. Fourteen species are recognized, two previously described species, T. wittei Benoît and T. suzannae Benoît, and twelve new: T. noorti sp. nov., T. denisi sp. nov., T. briani sp. nov., T. copelandi sp. nov., T. mourei sp. nov., T. lynnae sp. nov., T. normani sp. nov., T. luboi sp. nov., T. zuparkoi sp. nov., T. ohli sp. nov., T. camargoi sp. nov. and T. jimi sp. nov.. A key to species is proposed.

Three new species of Tuberepyris Lanes et Azevedo (Hymenoptera, Bethylidae), with amended diagnosis of the genus.

Three new species of the Afrotropical Tuberepyris are described and illustrated, T. nihilus Azevedo et Mugrabi, sp. nov. (Madagascar), T. codex Azevedo et Mugrabi, sp. nov. (South Africa), and T. hamus Azevedo et Mugrabi, sp. nov. (Central African Republic). The male of Tuberepyris is described for the first time. T. basibrevis Lanes et Azevedo from Tanzania is evaluated and the genus is redefined. A key to species of Tuberepyris is provided.

Revision of Thai Dissomphalus Ashmead, 1893 (Hymenoptera, Bethylidae), with description of twenty four new species.

The Thai Dissomphalus classification is reviewed, and 29 species are recognized. Previously described species, D. browni Terayama, D. chiangmaiensis Terayama, and D. thaianus Terayama had their known distribution and taxonomic variations broadened. Dissomphalus wusheanus Terayama is newly recorded from Thailand. Twenty-four new species, D. epitus Mugrabi & Azevedo, sp. nov., D. verus Mugrabi & Azevedo, sp. nov., D. prilus Mugrabi & Azevedo, sp. nov.,D. daneus Mugrabi & Azevedo, sp. nov., D. gabrus Mugrabi & Azevedo, sp. nov., D. gionus Mugrabi & Azevedo, sp. nov., D. cherrus Mugrabi & Azevedo, sp. nov., D. julius Mugrabi & Azevedo, sp. nov., D. davus Mugrabi & Azevedo, sp. nov., D. zethus Mugrabi & Azevedo, sp. nov., D. hetus Mugrabi & Azevedo, sp. nov., D. joelus Mugrabi & Azevedo, sp. nov., D. nandus Mugrabi & Azevedo, sp. nov., D. jubus Mugrabi & Azevedo, sp. nov., D. kelsus Mugrabi & Azevedo, sp. nov., D. turinus Mugrabi & Azevedo, sp. nov., D. paulus Mugrabi & Azevedo, sp. nov., D. barbus Mugrabi & Azevedo, sp. nov., D. robus Mugrabi & Azevedo, sp. nov., D. geanus Mugrabi & Azevedo, sp. nov., D. magnetus Mugrabi & Azevedo, sp. nov., D. lidinus Mugrabi & Azevedo, sp. nov., D. mugrus Mugrabi & Azevedo, sp. nov. and D. ferrerus Mugrabi & Azevedo, sp. nov., are described and illustrated. A key to the males of Dissomphalus from the Oriental region is provided.

Revision of Pristonesia (Hymenoptera, Bethylidae, Pristocerinae), an endemic Afrotropical genus.

The Afrotropical pristocerine genus Pristonesia is revised. Seven species are recognized, one previously described species, P. nyamuragira (Benoit), and six species are described and illustrated as new: P. oracil sp. nov., P. parcetil sp. nov., P. querfil sp. nov., P. sicril sp. nov., P. tainatril sp. nov., and P. uvenil sp. nov. A key to the Afrotropical species is presented. Additionally, the main morphological diagnostic characters and the morphological relationships of Pristonesia with other pristocerine genera are discussed. The diagnostic characters are revised, and the genus can be identified by the presence of apical projections on the harpe; a setose apical aedeagal lobe; a predominantly flattened cuspis; and the absence of a cupula on the ventral side of the genitalia.

Glutodon gen. nov. (Hymenoptera, Bethylidae), a flap-metasomed Pristocerinae from Madagascar.

Here we describe the flat wasp genus Glutodon gen. nov., and its single species G. mikeaensis sp. nov. from Madagascar. The males of this genus are easily recognized among the other genera of Pristocerinae by having the metasoma with a conical flap expansion on the sternum VIII and the hypopygium with the anterolateral apodeme very thick along its length with a very gibbous tip, both conditions unique in Bethylidae. Glutodon gen. nov. is the sixth genus of flat wasps endemic of Madagascar, and the 30th of Pristocerinae.

Synopsis of the Neotropical Sclerodermus Latreille (Hymenoptera, Bethylidae) with description of a new species attacking human beings.

We provide a Neotropical synopsis of the genus Sclerodermus Latreille. The genus is recognized by having the antenna with 11 flagellomeres, the head globoid or nearly so, the eyes present and flat, and the clypeal median lobe strongly angled and deeply separated from the lateral lobes. The strong polymorphism of having wings is an intra- and intergeneric feature and the macropterous specimens are diagnosed by having forewings with the pterostigma slender and only the R cell closed. In this synopsis we recognized seven previously known species: S. formicarius Kieffer, S. galapagensis Brues, S. iridomyrmicicola Bruch, S. macrogaster (Ashmead), S. soror Westwood, S. spilonotum Evans, and S. wilsoni Evans. The type-species, S. domesticus Latreille is also included, but their occurrence in the Neotropical Region is in doubt. Additionally, we discovered one more species, S. chicomendesi n. sp., which is described and illustrated, based on both sexes with molecular support. This species was found attacking a woman in her office in Porto Alegre, Brazil, where there was wood furniture with holes bored by beetles of the Ptinidae genus TricorynusWaterhouse. We also provide a key to the Neotropical species of this genus and a discussion about species attacking human beings.

Morphology and evolution of the mesopleuron in Bethylidae (Hymenoptera: Chrysidoidea) mapped on a molecular phylogeny.

The mesopleuron of Bethylidae has morphostructural characters that remain poorly understood, explored, and defined. The wide range of variability of this sclerite has generated confusion both in taxonomic and cladistic studies. Aiming to solve this issue, we describe the general mesopleural anatomy of Bethylidae. Our goal is to propose primary homologies by matching external and internal structures (muscles and apodemes). We reconstruct the ancestral state of the main mesopleural structures by applying the maximum-likelihood method on ten selected character states. The phylogeny of Bethylidae is built by analyzing a dataset of COI and 28S sequences, using maximum-likelihood and Bayesian inference methods. The Bethylidae and all subfamilies are recovered as monophyletic in both resulting phylogenies, with high clade support values. Although the two analyses yielded similar results, we used the tree resulting from the Bayesian inference to map the evolution of the morphological characters, as it is better supported. The study of mesopleural anatomy allows exploration and discussion of the evolution of characters and their present states in Bethylidae and Hymenoptera in general. Reconstruction of the ancestral states shows that many characters arose independently in Bethylidae subfamilies.

Floristic composition and similarity of 15 hectares in Central Amazon, Brazil.

The Amazon region is one of the most diverse areas in the world. Research on high tropical forest diversity brings up relevant contributions to understand the mechanisms that result and support such diversity. In the present study we describe the species composition and diversity of 15 one-ha plots in the Amazonian terra firme dense forest in Brazil, and compare the floristic similarity of these plots with other nine one-ha plots. The 15 plots studied were randomly selected from permanent plots at the Embrapa Experimental site, Amazonas State in 2005. The diversity was analysed by using species richness and Shannon's index, and by applying the Sorensen's index for similarity and unweighted pair-group average (UPGMA) as clustering method. Mantel test was performed to study whether the differences in species composition between sites could be explained by the geographic distance between them. Overall, we identified 8 771 individuals, 264 species and 51 plant families. Most of the species were concentrated in few families and few had large number of individuals. Families presenting the highest species richness were Fabaceae (Faboideae: 22spp., Mimosoideae: 22spp.), Sapotaceae: 22spp., Lecythidaceae: 15 and Lauraceae: 13. Burseraceae had the largest number of individuals with 11.8% of the total. The ten most abundant species were: Protium hebetatum (1 037 individuals), Eschweilera coriacea (471), Licania oblongifolia (310), Pouteria minima (293), Ocotea cernua (258), Scleronema micranthum (197), Eschweilera collina (176), Licania apelata (172), Naucleopsis caloneura (170) and Psidium araca (152), which represented 36.5% of all individuals. Approximately 49% of species had up to ten individuals and 13% appeared only once in all sampled plots, showing a large occurrence of rare species. Our study area is on a forest presenting a high tree species diversity with Shannon's diversity index of 4.49. The dendrogram showed two groups of plots with low similarity between them (less than 0.25), and the closer the plots were one to another, more similar in species composition (Mantel R = 0.3627, p < 0.01). The 15 plots in our study area share more than 50% of their species composition and represent the group of plots that have the shortest distance between each other. Overall, our results highlight the high local and regional heterogeneity of environments in terra firme forests, and the high occurrence of rare species, which should be considered in management and conservation programs in the Amazon rainforest, in order to maintain its structure on the long run.

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.

Revision of the world Apenesia Westwood (Hymenoptera, Bethylidae).

The world species of Apenesia are revised. Twenty-seven previously described species of Pristocerinae are addressed to Apenesia: A. amoena Evans, A. bicolor Vargas Terayama, A. chontalica Westwood, A. conradti Kieffer, A. delicata Evans, A. dominica Evans, A. flavipes Cameron, A. formosa Vargas Terayama, A. laevigata (Evans), A. levis Kieffer, A. leytensis (Terayama), A. makiharai (Sawada, Terayama Mita), A. malaitensis Brues, A. miki (Terayama), A. modesta (Smith), A. nigra Kieffer, A. parasitica (Smith), A. perlonga Corrêa Azevedo, A. proxima Kieffer, A. punctata Kieffer, A. sahyadrica Azevedo Waichert, A. singularis Lanes Azevedo, A. sjostedti (Tullgren), A. substriata Kieffer, A. unicolor Kieffer, and A. vaurieorum Evans. Additionally, 21 new species are described and illustrated: Apenesia amenula sp. nov.; A. aniela sp. nov.; A. azeda sp. nov.; A. beliella sp. nov.; A. berela sp. nov.; A. bifiela sp. nov.; A. celiela sp. nov.; A. chandela sp. nov.; A. cila sp. nov.; A. colombela sp. nov.; A. elela sp. nov.; A. esila sp. nov.; A. eura sp. nov.; A. farela sp. nov.; A. gabela sp. nov.; A. girena sp. nov.; A. goela sp. nov.; A. itoiela sp. nov.; A. joela sp. nov.; A. juliela sp. nov. and A. kelsiela sp. nov. The sexual association of A. celiela and A. azeda was possible due to biological data information. A key to species based on males is presented. The genus Apenesia is a parasitiod of beetles that live in galleries in dead wood or seeds.

DNA barcoding and hypopygium shape support delimitation of sympatric Dissomphalus species (Hymenoptera, Bethylidae) from the Atlantic rainforest.

Dissomphalus is a cosmopolitan genus of Bethylidae and has 269 Neotropical species divided into 32 species-groups, mostly defined by the genital and the tergal process structures. Dissomphalus rectilineus and D. concavatus are sympatric species in the ulceratus species-group. Members of the species-group share many similarities in the morphology of the head, hypopygium, tergal process and genitalia, but may be distinguished by the structure of the hypopygium. Previous studies have found intermediate structures of the hypopygium in the sympatric areas and raised questions about the distinctiveness of these two species. We sequenced 340 bp of the mitochondrial gene cytochrome oxidase I of 29 specimens from Brazil and Paraguay, calculated the genetic divergence among specimens, and recovered the phylogenetic relationships between taxa. In addition, we compared the morphology of the hypopygium to evaluate its use as a species-specific diagnostic character using the genetic divergence values. We recovered three well-supported monophyletic groups (intraclade divergence from 1.3 to 13.4%) and three hypopygium morphologies associated with each clade, two of them associated with D. rectilineus and D. concavatus (as described in the literature); the third one is new, not associated with any known species. The divergence between the D. rectilineus and D. concavatus clades was 19%, while the third clade is divergent from each species by 19-20%. If fully described, the hypopygium shape associated with the COI sequence will represent an extremely promising approach to the diagnosis of Dissomphalus species.

Biodiversity of the aculeate wasps (Hymenoptera: Aculeata) of the Arabian Peninsula: Chrysidoidea, Bethylidae.

An updated checklist of the Arabian species of Bethylidae is provided here. The list includes 29 identified valid species in 25 genera and five subfamilies: Bethylinae (2 genera, 0 species), Epyrinae (5 genera, 8 species), Mesitiinae (7 genera, 13 species), Pristocerinae (5 genus, 1 species) and Scleroderminae (6 genera, 7 species). Distributions, both on the Peninsula and extra-limital, are provided for each species; 17 species (59%) are endemic to the Arabian Peninsula.

Synopsis of world Discleroderma Kieffer (Hymenoptera, Bethylidae).

We provide a synopsis of the genus Discleroderma, which is known only from females. The genus is easily recognized by having tubercles on metasomal tergites III-V. In this synopsis we recognized D. gundari Terayama, D. indiensis Lanes Azevedo, D. tuberculatum Magretti, and D. yemenensis Lanes Azevedo as valid species. Additionally we discovered four more species D. concursum sp. nov., D. dolium sp. nov., D. plymouthoides sp. nov., and D. guamensis sp. nov., which are described and illustrated. We also provide an emended diagnosis and a key to the world species of this genus.

Revisited morphology applied for Systematics of flat wasps (Hymenoptera, Bethylidae).

The world fauna of the flat wasps (Bethylidae) is represented by about 3,000 valid species. The skeletal morphology of bethylids is still not adequately understood and the terminology is generally not standardized between its internal taxa and with other Hymenoptera families. The same scenario exists in most of the families in this order. To address this problem, we describe the external skeletal morphology of Bethylidae. We review the terms used to describe skeletal features in the Hymenoptera in general and a consensus terminology is proposed for Bethylidae, which is linked to the online Hymenoptera Anatomy Ontology. The morphology of the studied specimens is illustrated with photos and line drawings. We also discuss the morphological variation at both subfamilial and generic ranks. Our analyses challenge hundreds of inappropriate, confused or imprecise terms traditionally used for Hymenoptera morphology. As a result, we have applied hundreds of updates of the terminology available online at the Hymenoptera Anatomy Ontology.

Review of Afrotropical Glenosema Kieffer (Hymenoptera, Bethylidae) with description of 13 new species.

The Afrotropical species of Glenosema are reviewed. Seventeen species are recognized, four previously described species, G. niloticum (Kieffer), G. durum Barbosa, Kawada Azevedo, G. denteatum Lanes Azevedo and G. elevatum Lanes Azevedo, and thirteen species described and illustrated as new: G. qaqambile sp. nov., G. deki sp. nov., G. ebazileyo sp. nov., G. tyaba sp. nov., G. umphanda sp. nov., G. banzi sp. nov., G. bubanzi sp. nov., G. ahlukileyo sp. nov., G. icephe sp. nov., G. mxinwa sp. nov., G. lingana sp. nov., G. uphahla sp. nov. and G. tyheli sp. nov. A key to the Afrotropical species is presented. We highlight potential character suites that may be phylogenetically significant and consider evolutionary scenarios that may have led to the present distribution pattern associated with semi-arid areas on the African continent.

The Forest Observation System, building a global reference dataset for remote sensing of forest biomass.

Forest biomass is an essential indicator for monitoring the Earth's ecosystems and climate. It is a critical input to greenhouse gas accounting, estimation of carbon losses and forest degradation, assessment of renewable energy potential, and for developing climate change mitigation policies such as REDD+, among others. Wall-to-wall mapping of aboveground biomass (AGB) is now possible with satellite remote sensing (RS). However, RS methods require extant, up-to-date, reliable, representative and comparable in situ data for calibration and validation. Here, we present the Forest Observation System (FOS) initiative, an international cooperation to establish and maintain a global in situ forest biomass database. AGB and canopy height estimates with their associated uncertainties are derived at a 0.25 ha scale from field measurements made in permanent research plots across the world's forests. All plot estimates are geolocated and have a size that allows for direct comparison with many RS measurements. The FOS offers the potential to improve the accuracy of RS-based biomass products while developing new synergies between the RS and ground-based ecosystem research communities.

Revision of Anisepyris Kieffer (Hymenoptera, Bethylidae), with description of 135 new species.

The identity of many species of Anisepyris Kieffer is currently unknown, mostly because the available taxonomic data are not sufficiently accurate to allow for precise identifications. To solve this problem, all 121 described species were investigated, including the analyses of 115 holotypes and 26 allotypes. Almost all Neotropical species of Epyrinae and many from the Nearctic were also examined in order to recognize possible species of Anisepyris misclassified in other genera. The ten species-groups included in Anisepyris are redefined. And the female genital structures of the genus are investigated for the first time. Based on this, we recognize 119 previously described species as belonging to Anisepyris, one species is transferred to Laelius Ashmead, L. foveaticeps (Rosmann Azevedo) comb. nov., one is transferred to Chlorepyris Kieffer, C. nigerrimus (Evans) comb. nov., and 135 new species are described to science, so that the genus is now contains 254 species. They are recorded from 31 countries of the New World. Lectotypes are designated for A. aurichalceus (Westwood) and A. planiceps (Fabricius). The previously identified species-groups are re-organized into six, and seven additional species-groups are proposed are new. Identification keys for Anisepyris species-groups and species are provided.

Pairs in copulation of the highly dimorphic genus Pristocera Klug (Hymenoptera, Bethylidae) from Madagascar solve taxonomic problems of male-female associations.

The main goal of this paper is to update the taxonomic information of the pairs of Pristocera Klug collected in Madagascar by A. Seyrig. Within this material twelve species are recognized, ten are new to science, all based on pairs collected in copulation, which are described and illustrated: P. melmani sp. nov., P. alexi sp. nov., P. martyi sp. nov., P. gloriae sp. nov., P. julieni sp. nov., P. morti sp. nov., P. mauricei sp. nov., P. makungai sp. nov., P. teetsii sp. nov. and P. zubai sp. nov.. Pristocera levicollis (Kieffer) had its female discovered, and P. cambouei Saussure stat. rev. had its lectotype designated. Sclerodermus hova Saussure is transferred back to Pristocera and considered as a junior synonym of P. cambouei. Keys for males and females to the species of Pristocera from Madagascar are provided. The Malagasy fauna of Pristocera is now represented by twelve species, all endemic to this island.

New species and records of Dissomphalus Ashmead (Hymenoptera, Bethylidae) from Cerrado, Caatinga and relicts of the Atlantic Forest from northeastern Brazil.

The main goal of this paper is to study the Dissomphalus material deposited at the Coleção Zoológica do Maranhão (Maranhão, Brazil). The material was collected in six parks: Parque Nacional Chapada das Mesas, Parque Estadual do Mirador and Reserva Ecológica do Inhamum in the state of Maranhão; Parque Nacional Serra das Confusões and Parque Nacional de Sete Cidades in the state of Piauí, and Parque Nacional de Ubajara in the state of Ceará. Forty species are recognized from these six parks. Twenty-two of them are previously described species, namely: D. bifurcatus Azevedo, D. cervoides Azevedo, D. conicus Azevedo, D. curvilongus Azevedo, D. geniculatus Azevedo, D. gilvipes Evans, D. infissus Evans, D. krombeini Azevedo, D. laminaris Redighieri Azevedo, D. latimerus Azevedo, D. linearis Azevedo, D. lobisserratus Azevedo, D. magnus Redighieri Azevedo, D. megomphalus Evans, D. microstictus Evans, D. napo Evans, D. plaumanni Evans, D. scamatus Azevedo, D. subpilosus Azevedo, D. tubulatus Redighieri Azevedo, D. ubracus Brito Azevedo and D. verrucosus Redighieri Azevedo. Eighteen species are newly described and illustrated: Dissomphalus acutipupu Colombo Azevedo sp. nov., D. boibumba Colombo Azevedo sp. nov., D. boitata Colombo Azevedo sp. nov., D. boiuna Colombo Azevedo sp. nov., D. boto Colombo Azevedo sp. nov., D. caipora Colombo Azevedo sp. nov., D. capelobo Colombo Azevedo sp. nov., D. cuca Colombo Azevedo sp. nov., D. curupira Colombo Azevedo sp. nov., D. iara Colombo Azevedo sp. nov., D. iracema Colombo Azevedo sp. nov., D. irupe Colombo Azevedo sp. nov., D. joaquinae Colombo Azevedo sp. nov., D. jurupari Colombo Azevedo sp. nov., D. mapinguari Colombo Azevedo sp. nov., D. matintaperera Colombo Azevedo sp. nov., D. saci Colombo Azevedo sp. nov. and D. torem Colombo Azevedo sp. nov.. A key for these species is provided. The genus is newly recorded from the state of Piauí, Brazil.