Pollen DNA metabarcoding reveals cryptic diversity and high spatial turnover in alpine plant-pollinator networks.

Alpine plant-pollinator communities play an important role in the functioning of alpine ecosystems, which are highly threatened by climate change. However, we still have a poor understanding of how environmental factors and spatiotemporal variability shape these communities. Here, we investigate what drives structure and beta diversity in a plant-pollinator metacommunity from the Australian alpine region using two approaches: pollen DNA metabarcoding (MB) and observations. Individual pollinators often carry pollen from multiple plant species, and therefore we expected MB to reveal a more diverse and complex network structure. We used two gene regions (ITS2 and trnL) to identify plant species present in the pollen loads of 154 insect pollinator specimens from three alpine habitats and construct MB networks, and compared them to networks based on observations alone. We compared species and interaction turnover across space for both types of networks, and evaluated their differences for plant phylogenetic diversity and beta diversity. We found significant structural differences between the two types of networks; notably, MB networks were much less specialized but more diverse than observation networks, with MB detecting many cryptic plant species. Both approaches revealed that alpine pollination networks are very generalized, but we estimated a high spatial turnover of plant species (0.79) and interaction rewiring (0.6) as well as high plant phylogenetic diversity (0.68) driven by habitat differences based on the larger diversity of plant species and species interactions detected with MB. Overall, our findings show that habitat and microclimatic heterogeneity drives diversity and fine-scale spatial turnover of alpine plant-pollinator networks.

Dominance of coniferous and broadleaved trees drives bacterial associations with boreal feather mosses.

The composition of ecologically important moss-associated bacterial communities seems to be mainly driven by host species but may also be shaped by environmental conditions related with tree dominance. The moss phyllosphere has been studied in coniferous forests while broadleaf forests remain understudied. To determine if host species or environmental conditions defined by tree dominance drives the bacterial diversity in the moss phyllosphere, we used 16S rRNA gene amplicon sequencing to quantify changes in bacterial communities as a function of host species (Pleurozium schreberi and Ptilium crista-castrensis) and forest type (coniferous black spruce versus deciduous broadleaf trembling aspen) in eastern Canada. The overall composition of moss phyllosphere was defined by the interaction of both factors, though most of the bacterial phyla were determined by a strong effect of forest type. Bacterial α-diversity was highest in spruce forests, while there was greater turnover (ß-diversity) and higher γ-diversity in aspen forests. Unexpectedly, Cyanobacteria were much more relatively abundant in aspen than in spruce forests, with the cyanobacteria family Nostocaceae differing the most between forest types. Our results advance the understanding of moss-associated microbial communities among coniferous and broadleaf deciduous forests, which are important with the increasing changes in tree dominance in the boreal system.

Phylogeography and evolution of the Red Salamander (Pseudotriton ruber).

Phylogeographic studies frequently result in the elevation of subspecific taxa to species given monophyly, or the synonymy of subspecies that are not monophyletic. However, given limited or incongruent datasets, retention of subspecies can be useful to describe hypothesized incipient species or to illustrate interesting biological phenomena driving morphological diversity. Four subspecific taxa have been used to describe largely allopatric geographic variation within the species Pseudotriton ruber, a plethodontid salamander occupying stream and spring habitats across eastern North America: P. r. vioscai occurs in lowland Coastal Plain habitats, while P. r. ruber, P. r. nitidus, and P. r. schencki occupy upland regions in and around the Appalachian Mountains. Pseudotriton ruber co-occurs through its distribution with the aposematic newt Notophthalmus viridescens, and both species are hypothesized to be part of a Müllerian mimicry complex. In this study, we sequenced regions of two mitochondrial (cytochrome b, NADH dehydrogenase subunit 2) and one single copy nuclear protein-coding gene (pro-opiomelanocortin) from individuals sampled across much of the distribution of P. ruber and then used maximum-likelihood and Bayesian phylogenetic inference to test the monophyly of subspecies, reconstruct biogeographic history, and make inferences about morphological evolution. Phylogeographic hypotheses from mitochondrial and nuclear datasets described structure among populations of P. ruber which separated Coastal Plain and upland Appalachian populations, but subspecies were not monophyletic. Biogeographic reconstruction estimated the ancestor of all populations to have occupied and initially diverged in the Coastal Plain during the Pliocene (∼3.6mya), before one lineage subsequently invaded upland areas of Appalachia. Bold bright coloration of high elevation subspecies P. r. nitidus and P. r. schencki appears to have evolved twice. We hypothesize that the Müllerian mimicry complex with N. viridescens and P. ruber may provide a selective mechanism driving the co-evolution of striking bright and dull morphological variation among populations of both species. While P. ruber subspecies were not consistent with our criteria for diagnosing species (monophyly) and therefore could not be elevated to species, we advocate for the retention of subspecies because they describe hypotheses about an incipient species (P. r. vioscai) and how Müllerian mimicry may shape morphological diversity of species.

Molecular phylogeny of Pompilinae (Hymenoptera: Pompilidae): Evidence for rapid diversification and host shifts in spider wasps.

Pompilinae is one of the largest subfamilies of spider wasps (Pompilidae). Most pompilines are generalist spider predators at the family level, but some taxa exhibit ecological specificity (i.e., to spider-host guild). Here we present the first molecular phylogenetic analysis of Pompilinae, toward the aim of evaluating the monophyly of tribes and genera. We further test whether changes in the rate of diversification are associated with host-guild shifts. Molecular data were collected from five nuclear loci (28S, EF1-F2, LWRh, Wg, Pol2) for 76 taxa in 39 genera. Data were analyzed using maximum likelihood (ML) and Bayesian inference (BI). The phylogenetic results were compared with previous hypotheses of subfamilial and tribal classification, as well as generic relationships in the subfamily. The classification of Pompilus and Agenioideus is also discussed. A Bayesian relaxed molecular clock analysis was used to examine divergence times. Diversification rate-shift tests accounted for taxon-sampling bias using ML and BI approaches. Ancestral host family and host guild were reconstructed using MP and ML methods. Ancestral host guild for all Pompilinae, for the ancestor at the node where a diversification rate-shift was detected, and two more nodes back in time was inferred using BI. In the resulting phylogenies, Aporini was the only previously proposed monophyletic tribe. Several genera (e.g., Pompilus, Microphadnus and Schistonyx) are also not monophyletic. Dating analyses produced a well-supported chronogram consistent with topologies from BI and ML results. The BI ancestral host-use reconstruction inferred the use of spiders belonging to the guild "other hunters" (frequenting the ground and vegetation) as the ancestral state for Pompilinae. This guild had the highest probability for the ML reconstruction and was equivocal for the MP reconstruction; various switching events to other guilds occurred throughout the evolution of the group. The diversification of Pompilinae shows one main rate-shift coinciding with a shift to ground-hunter spiders, as reconstructed by the BI ancestral character-state analysis.

A case control study of the implementation of change model versus passive dissemination of practice guidelines for compliance in monitoring for metabolic syndrome.

We developed an intervention to improve compliance with guidelines for monitoring metabolic syndrome and compared compliance prior to intervention and three times post-intervention at three community mental health clinics in Texas. One test clinic received intervention and two other clinics served as controls. Fifty random charts were reviewed from each clinic for three specific, 1-2 weeks periods over the course of 18 months. There were significant improvements in the ordering of labs, the presence of lab results in the chart, and documentation of blood pressure, body mass index and waist circumference in the intervention clinic over time in comparison to the control clinics. Documented evidence of physician action with respect to out of range values remained low. Metabolic monitoring is a multi-step process. Removing barriers, creating specific procedures, and dedicating staff resources can improve compliance with monitoring.

Functional and Proteomic Insights into Aculeata Venoms.

Aculeate hymenopterans use their venom for a variety of different purposes. The venom of solitary aculeates paralyze and preserve prey without killing it, whereas social aculeates utilize their venom in defence of their colony. These distinct applications of venom suggest that its components and their functions are also likely to differ. This study investigates a range of solitary and social species across Aculeata. We combined electrophoretic, mass spectrometric, and transcriptomic techniques to characterize the compositions of venoms from an incredibly diverse taxon. In addition, in vitro assays shed light on their biological activities. Although there were many common components identified in the venoms of species with different social behavior, there were also significant variations in the presence and activity of enzymes such as phospholipase A2s and serine proteases and the cytotoxicity of the venoms. Social aculeate venom showed higher presence of peptides that cause damage and pain in victims. The venom-gland transcriptome from the European honeybee (Apis mellifera) contained highly conserved toxins which match those identified by previous investigations. In contrast, venoms from less-studied taxa returned limited results from our proteomic databases, suggesting that they contain unique toxins.

Nest Ecology and Prey Preference of the Mud Dauber Wasp Sceliphron formosum (Hymenoptera: Sphecidae).

(1) Background: Sceliphron is a genus of sphecid wasps that build mud nests for reproduction. These wasps prey exclusively on spiders, and commonly inhabit human constructions. The nesting behaviour and prey selection of many Sceliphron species are well studied, but despite being a common insect in urban areas, Sceliphron formosum has never been comprehensively studied. (2) Methods: In this study, over 650 mud nests of S. formosum were collected, analysed and examined to establish prey preference, nest ecology and interspecific interactions. Prey preference was evaluated in terms of abundance, diversity and morphology. Preference in terms of morphology was estimated using body length to leg span ratio (BLR). (3) Results: S. formosum largely preys on ground-hunting spiders, among which Salticidae represented the most collected prey. In terms of prey size, S. formosum captures prey with a large BLR. Moreover, an unexpected discovery showed that the enclosed mud nests provide a micro niche that supports a wide variety of insects. Sixteen families and 23 species of insects were found associated with the use of mud nests, comprising the insect orders Hymenoptera, Diptera and Coleoptera. These included important pollinators, new species and native species not recorded in the past 20 years of mud dauber wasp research. We propose the potential of S. formosum as a keystone species, due to its ability to provide a micro niche for native species in urban areas. We also discuss how these results contribute to our knowledge on the role of insects in urban ecosystems and their significance in relation to conservation, ecology and biodiversity studies.

Twenty two years later: An updated checklist of Neotropical spider wasps (Hymenoptera: Pompilidae).

Twenty two years after the first checklist of Neotropical Spider Wasps, a new list of genera and species is offered, including novelties in phylogeny and systematics, as well as reviews, synonyms and descriptions since the year 2000. Sixty three genera and 946 species of Pompilidae are listed.

GoCC4All a pervasive technology to provide access to TV to the deafblind community.

Considering the importance of communication and independence for the deafblind community, this work presents findings of the use of technology to address the lack of information due to communication challenges among the deafblind community. Over time, many investigations have been carried out regarding this matter, but very few providing solution, which is why this study emerged, looking to making all the information broadcasted through television accessible for this community. The work team designed a technology (GoCC4All) to address the needs of the deafblind community. GoCC4All provides access to captions available on TV through braille displays and mobile devices. Our research process and results outline the path for creating, adapting, and adopting new technologies for people with disabilities who have the right to access the information just as their peers without disabilities. The information in this paper is based on two surveys, an initial beta testing (BT) and a final survey among a group of 14 users (UT) who tested the GoCC4All application. Our findings support the positive impact of the iterative creation of assistive technology based on users' experience and users' recommendations to better serve the needs of the deafblind community.

A Short Review of the Venoms and Toxins of Spider Wasps (Hymenoptera: Pompilidae).

Parasitoid wasps represent the plurality of venomous animals, but have received extremely little research in proportion to this taxonomic diversity. The lion's share of investigation into insect venoms has focused on eusocial hymenopterans, but even this small sampling shows great promise for the development of new active substances. The family Pompilidae is known as the spider wasps because of their reproductive habits which include hunting for spiders, delivering a paralyzing sting, and entombing them in burrows with one of the wasp's eggs to serve as food for the developing larva. The largest members of this family, especially the tarantula hawks of the genus Pepsis, have attained notoriety for their large size, dramatic coloration, long-term paralysis of their prey, and incredibly painful defensive stings. In this paper we review the existing research regarding the composition and function of pompilid venoms, discuss parallels from other venom literatures, identify possible avenues for the adaptation of pompilid toxins towards human purposes, and future directions of inquiry for the field.

Detection of the Japanese encephalitis vector mosquito Culex tritaeniorhynchus in Australia using molecular diagnostics and morphology.

BACKGROUND: Culex (Culex) tritaeniorhynchus is an important vector of Japanese encephalitis virus (JEV) affecting feral pigs, native mammals and humans. The mosquito species is widely distributed throughout Southeast Asia, Africa and Europe, and thought to be absent in Australia. METHODS: In February and May, 2020 the Medical Entomology unit of the Northern Territory (NT) Top End Health Service collected Cx. tritaeniorhynchus female specimens (n = 19) from the Darwin and Katherine regions. Specimens were preliminarily identified morphologically as the Vishnui subgroup in subgenus Culex. Molecular identification was performed using cytochrome c oxidase subunit 1 (COI) barcoding, including sequence percentage identity using BLAST and tree-based identification using maximum likelihood analysis in the IQ-TREE software package. Once identified using COI, specimens were reanalysed for diagnostic morphological characters to inform a new taxonomic key to related species from the NT. RESULTS: Sequence percentage analysis of COI revealed that specimens from the NT shared 99.7% nucleotide identity to a haplotype of Cx. tritaeniorhynchus from Dili, Timor-Leste. The phylogenetic analysis showed that the NT specimens formed a monophyletic clade with other Cx. tritaeniorhynchus from Southeast Asia and the Middle East. We provide COI barcodes for most NT species from the Vishnui subgroup to aid future identifications, including the first genetic sequences for Culex (Culex) crinicauda and the undescribed species Culex (Culex) sp. No. 32 of Marks. Useful diagnostic morphological characters were identified and are presented in a taxonomic key to adult females to separate Cx. tritaeniorhynchus from other members of the Vishnui subgroup from the NT. CONCLUSIONS: We report the detection of Cx. tritaeniorhynchus in Australia from the Darwin and Katherine regions of the NT. The vector is likely to be already established in northern Australia, given the wide geographical spread throughout the Top End of the NT. The establishment of Cx. tritaeniorhynchus in Australia is a concern to health officials as the species is an important vector of JEV and is now the sixth species from the subgenus Culex capable of vectoring JEV in Australia. We suggest that the species must now be continuously monitored during routine mosquito surveillance programmes to determine its current geographical spread and prevent the potential transmission of exotic JEV throughout Australia.

The first true millipede-1306 legs long.

The name "millipede" translates to a thousand feet (from mille "thousand" and pes "foot"). However, no millipede has ever been described with more than 750 legs. We discovered a new record-setting species of millipede with 1,306 legs, Eumillipes persephone, from Western Australia. This diminutive animal (0.95 mm wide, 95.7 mm long) has 330 segments, a cone-shaped head with enormous antennae, and a beak for feeding. A distant relative of the previous record holder, Illacme plenipes from California, it belongs to a different order, the Polyzoniida. Discovered 60 m below ground in a drill hole created for mineral exploration, E. persephone possesses troglomorphic features; it lacks eyes and pigmentation, and it has a greatly elongated body-features that stand in stark contrast to its closest surface-dwelling relatives in Australia and all other members of its order. Using phylogenomics, we found that super-elongation (> 180 segments) evolved repeatedly in the millipede class Diplopoda. The striking morphological similarity between E. persephone and I. plenipes is a result of convergent evolution, probably for locomotion in similar soil habitats. Discovered in the resource-rich Goldfields-Esperance region and threatened by encroaching surface mining, documentation of this species and conservation of its habitat are of critical importance.

A new genus and species of narrow-range millipede (Diplopoda, Polydesmida, Dalodesmidae) from Tasmania, Australia.

Kebodesmuszonarius gen. nov. et sp. nov. is only known from a small area on the Great Western Tiers in northern Tasmania, Australia, and like species of Paredrodesmus Mesibov, 2003 has no detectable paranota on the diplosegments. The gonopod telopodite of the new species is divided into a large, lateral, cowl-like structure, a solenomere and a medial branch with three processes.

Three new species of Epipompilus Kohl (Hymenoptera, Pompilidae, Pepsinae) from Australia.

Three new species are added to the genus Epipompilus (Hymenoptera: Pompilidae) in Australia. Epipompilus mirabundus sp. nov., E. taree sp. nov., and E. namadgi sp. nov. are described and illustrated. A key to males of Epipompilus is provided. A novel association of Epipompilus and Sceliphron formosum (Hymenoptera: Sphecidae) is also documented. The larva of E. mirabundus sp. nov. was found sharing single nest cell with a Sceliphron larva; this association could be the result of a parasitised spider being brought back to the nest by the Sceliphron adult.

Re-examination of Australian and Oriental species of Minagenia Banks, 1934 (Hymenoptera: Pompilidae), with a new record for the genus in Australia and a new species description.

The genus Minagenia Banks, 1934 is reported for the first time from Australia. A new species, M. lasallei Decker, Rodriguez Pitts, sp. nov. (Australia: from Queensland to Tasmania), is described and illustrated. A key to the four species from the Australasian and Oriental regions is given. Three species are removed from Asian Minagenia: granulosa Tsuneki, 1989, comb. nov., to the genus Priocnemis Schiødte, 1837; pempuchiensis Tsuneki, 1989, comb. nov., to the genus Nipponodipogon Ishikawa, 1965; and assimilis Smith, 1861, comb. nov., to the genus Dichragenia Haupt, 1950. Minagenia alticola Tsuneki, 1989 is a member of an undescribed genus. Placement of this species is maintained in Minagenia until the new genus can be described in a future work. Minagenia brevicornis (Banks, 1934) is redescribed.

Ecological Drivers and Sex-Based Variation in Body Size and Shape in the Queensland Fruit Fly, Bactrocera tryoni (Diptera: Tephritidae).

The Queensland fruit fly (Bactrocera tryoni; Q-fly) is an Australian endemic horticultural pest species, which has caused enormous economic losses. It has the potential to expand its range to currently Q-fly-free areas and poses a serious threat to the Australian horticultural industry. A large number of studies have investigated the correlation between environmental factors and Q-fly development, reproduction, and expansion. However, it is still not clear how Q-fly morphological traits vary with the environment. Our study focused on three morphological traits (body size, wing shape, and fluctuating asymmetry) in Q-fly samples collected from 1955 to 1965. We assessed how these traits vary by sex, and in response to latitude, environmental variables, and geographic distance. First, we found sexual dimorphism in body size and wing shape, but not in fluctuating asymmetry. Females had a larger body size but shorter and wider wings than males, which may be due to reproductive and/or locomotion differences between females and males. Secondly, the body size of Q-flies varied with latitude, which conforms to Bergmann's rule. Finally, we found Q-fly wing shape was more closely related to temperature rather than aridity, and low temperature and high aridity may lead to high asymmetry in Q-fly populations.

Sand wasp (Hymenoptera: Crabronidae) parasites emerging from mud wasp nests (Hymenoptera: Sphecidae)-a reliable host record of Thraxan Yeates amp; Lambkin (Diptera: Bombyliidae: Anthracinae) with description of the pupal exuviae of three Thraxan species.

Two female Thraxan sp. (Diptera: Bombyliidae: Anthracinae) emerged from parasitised Pison simillimum Smith (Hymenoptera: Crabronidae) larvae found inside a Sceliphron formosum Smith (Hymenoptera: Sphecidae) mud wasps nest. Thraxan sp. larvae are endoparasites of P. simillimum larvae. The endoparasite pupates inside the host larva, kills it, and emerges free of the host body. We describe and illustrate the pupal exuviae of the newly found species and two known species, T. luteus Yeates Lambkin and T. misatulus Yeates Lambkin, and we compare these pupal exuviae to those of Anthrax Scopoli.

Pollinator size and its consequences: Robust estimates of body size in pollinating insects.

Body size is an integral functional trait that underlies pollination-related ecological processes, yet it is often impractical to measure directly. Allometric scaling laws have been used to overcome this problem. However, most existing models rely upon small sample sizes, geographically restricted sampling and have limited applicability for non-bee taxa. Allometric models that consider biogeography, phylogenetic relatedness, and intraspecific variation are urgently required to ensure greater accuracy. We measured body size as dry weight and intertegular distance (ITD) of 391 bee species (4,035 specimens) and 103 hoverfly species (399 specimens) across four biogeographic regions: Australia, Europe, North America, and South America. We updated existing models within a Bayesian mixed-model framework to test the power of ITD to predict interspecific variation in pollinator dry weight in interaction with different co-variates: phylogeny or taxonomy, sexual dimorphism, and biogeographic region. In addition, we used ordinary least squares regression to assess intraspecific dry weight ~ ITD relationships for ten bees and five hoverfly species. Including co-variates led to more robust interspecific body size predictions for both bees and hoverflies relative to models with the ITD alone. In contrast, at the intraspecific level, our results demonstrate that the ITD is an inconsistent predictor of body size for bees and hoverflies. The use of allometric scaling laws to estimate body size is more suitable for interspecific comparative analyses than assessing intraspecific variation. Collectively, these models form the basis of the dynamic R package, "pollimetry," which provides a comprehensive resource for allometric pollination research worldwide.

A new species of Carvalhomiris from Colombia with an assessment of its phylogenetic position (Heteroptera, Miridae, Orthotylinae).

Plant bugs, species of Miridae (Heteroptera), are not well known in the Neotropics, and Colombia is not an exception. Based on data from the available systematic catalog (Schuh 2002-2013) fewer than 150 species are recorded from the country, clearly an underestimation. Recent fieldwork has resulted in several new interesting taxa from Colombia. Carvalhomiris Maldonado & Ferreira, 1971, contains three described species from Colombia and Ecuador. From specimens collected in Jardín, Antioquia, Carvalhomirishenryi sp. n. is described. Images of the dorsal habitus and the male and female genitalia are provided. Based on morphological examination of the new species and published information, morphological characters were coded to construct a phylogenetic matrix for a cladistic analysis in which the phylogenetic position of the new species is assessed. Carvalhomirishenryi sp. n. is the northernmost species of the genus and noteworthy because it is the first record of any species of the genus in the Western Cordillera: all other species are known from the eastern flank of the Andes (Ecuador) or the Eastern Cordillera (Colombia). Natural history observations of the new species, including associations with composites (Asteraceae) are provided. It is speculated that the mirid might be predacious.

Phylogeny and population genetic analyses reveals cryptic speciation in the Bombus fervidus species complex (Hymenoptera: Apidae).

Bumble bees (Bombus Latrielle) are significant pollinators of flowering plants due to their large body size, abundant setae, and generalist foraging strategies. However, shared setal coloration patterns among closely and distantly related bumble bee species makes identification notoriously difficult. The advent of molecular genetic techniques has increased our understanding of bumble bee evolution and taxonomy, and enables effective conservation policy and management. Individuals belonging to the North American Bombus fervidus species-complex (SC) are homogenous in body structure but exhibit significant body color phenotype variation across their geographic distribution. Given the uncertainty of the genealogical boundaries within the SC, some authors have synonymized all members of the B. fervidus SC within a single taxon, while others propose an alternative two taxa hypothesis. Operating under the phylogenetic species concept, our analysis supports the hypothesis that there are two independent lineages of bumble bees within the B. fervidus SC. With the current evidence, however, it is not possible to assign valid names to either of them, because both lineages include the color phenotypes found in the original species descriptions of B. fervidus and B. californicus. Cryptic speciation does not seem to be the product of Müllerian mimicry between the clades, because diverging coloration patterns are observed when the distribution of the clades overlaps. Furthermore, within each lineage there is evidence for strong population differentiation that is correlated with geographic distribution rather than color phenotype. In our study, we demonstrate the importance of obtaining a broad sample of multiple populations when conducting lower-level phylogenetic analyses. In addition to improving our knowledge of bumble bee diversification patterns, characterizing the evolutionary history of these pollinators provides the foundation needed to guide contemporary conservation assessments and management strategies.