Review of the European Eumenes Latreille (Hymenoptera, Vespidae) using morphology and DNA barcodes, with an illustrated key to species.

The European species of the potter wasp genus Eumenes Latreille, 1802 (Vespidae, Eumeninae) are illustrated and a new illustrated key to the 13 recognised species is presented. Eumenesmediterraneusaemilianus Guiglia, 1951 is synonymised with E.papillarius (Christ, 1791) (syn. nov.), E.obscurus André, 1884 and E.andrei Dalla Torre, 1894 with E.pedunculatus (Panzer, 1799) (syn. nov.) and E.crimensis Blüthgen, 1938 with E.sareptanus André, 1884 (syn. nov.).

A new species of the genus Conosiphon Becker, 1923 and the first records of this genus for Europe (Diptera, Asilidae).

A new species of Conosiphon Becker, 1923, Conosiphonianus Álvarez Fidalgo & van den Broek, sp. nov., is described from Spain, representing the first record of this genus for Europe. It is illustrated in high-resolution photographs and the first ecological information is provided, as well as a key to all species tentatively placed in this genus.

A new species of the genus Psilota Meigen, 1822 (Diptera: Syrphidae) from China.

The genus Psilota Meigen, 1822 is recorded for the first time from China, and the species Psilota bashanensis Huo and Zhao sp. nov. is described and illustrated based on the adult male. The complete cytochrome c oxidase subunit I (COI) gene of this new species has been successfully obtained and compared to that of other congeneric species. An updated key to adult males of the genus Psilota from the Palaearctic Region is also provided.

A pictorial key for identification of the hoverflies (Diptera: Syrphidae) of the Madeira Archipelago.

Background: Syrphid flies are important ecological indicators and provide crucial ecosystem services, being important pollinators and biological control agents of insect pests. These charismatic insects are conspicuous and, due to their size and colourful patterns, are relatively easy to identify. However, the lack of user-friendly literature (e.g. photographic guides) for most areas may hamper its wider selection as a study group in biodiversity and ecological studies. The syrphid fauna of Madeira Archipelago comprises 26 species, including four endemics (Eumerushispidus Smit, Aguiar & Wakeham-Dawson, 2004; Melanostomawollastoni Wakeham-Dawson, Aguiar, Smit, McCullough & Wyatt, 2004; Myathropausta, Wollaston, 1858 and Xanthandrusbabyssa, Walker, 1849), but, despite the current good taxonomic knowledge on this group, information on species distribution, ecology and conservation is still lacking. Here, we provide a pictorial key to the adult hoverflies of Madeira Archipelago highlighting diagnostic characteristics and present photographs of both males and females (in dorsal and lateral views) in colour plates. The key and plates will help researchers to differentiate these species, thus encouraging the use of this insect group in future bioindication studies. In addition, this study also aims to engage a broader audience of non-experts in improving the knowledge on the distribution and ecology of Madeira syrphids. New information: We provide a checklist for the hoverflies of Madeira Archipelago and a pictorial key to help on species identification.

The European Stag Beetle (Lucanus cervus) Monitoring Network: International Citizen Science Cooperation Reveals Regional Differences in Phenology and Temperature Response.

To address the decline in biodiversity, international cooperation in monitoring of threatened species is needed. Citizen science can play a crucial role in achieving this challenging goal, but most citizen science projects have been established at national or regional scales. Here we report on the establishment and initial findings of the European Stag Beetle Monitoring Network (ESBMN), an international network of stag beetle (Lucanus cervus) monitoring schemes using the same protocol. The network, started in 2016, currently includes 14 countries (see results) but with a strong variation in output regarding the number of transects (148 successful transects in total) and transect walks (1735). We found differences across European regions in the number of stag beetles recorded, related to phenology and temperature, but not for time of transect start. Furthermore, the initial experiences of the ESBMN regarding international cooperation, citizen science approach, and drop-out of volunteers is discussed. An international standardised protocol that allows some local variation is essential for international collaboration and data management, and analysis is best performed at the international level, whereas recruiting, training, and maintaining volunteers is best organised locally. In conclusion, we appeal for more joint international citizen science-based monitoring initiatives assisting international red-listing and conservation actions.

The fruit flies (Diptera, Tephritidae) of the Madeira archipelago with the description of a new Oedosphenella Frey.

A checklist of the tephritid fauna (Diptera: Tephritidae) from Madeira archipelago is presented, including 17 known species and new distribution records for Porto Santo and Desertas Islands. An identification key to Madeira Tephritidae species with detailed illustrations is also provided. A new species of Oedosphenella Frey 1936, discovered in Madeira Island in 2004, is described as Oedosphenella bob sp. n. This genus is redefined and a key to the known species is provided. Comparative morphological analysis revealed that Bevismyia basuto Munro 1957 from Malawi should be included in the genus Oedosphenella. Therefore, we consider the genus name Oedosphenella Frey 1936 to be a senior synonym of Bevismyia Munro 1957 syn.n.

Three new hoverfly species from Greece (Diptera: Syrphidae).

An ongoing investigation on the Greek hoverfly fauna using adult morphology has revealed new species within three genera. In this study, the knowledge of the Mediterranean hoverfly fauna has been enhanced by describing the following species: Cheilosia candida Vujic et Radenkovic sp. n. (Pindos Mountains), Paragus thracusi Radenkovic, Likov et Vujic sp. n. (Rhodope Mountains) and Psilota aegeae Vujic, Ståhls et Smit sp. n. (Lesvos island). Diagnosis of new species, as well as identification keys to the Mediterranean species of the subgenus Convocheila Barkalov of Cheilosia Meigen and the European species of the genus Psilota Meigen have been provided. Additionally, mtDNA COI barcodes for the members of the Psilota atra group (except Psilota nana Smit et Vujic) have been given. In addition, the taxonomic status of Psilota anthracina Meigen has been discussed.

A new species of Eumerus (Diptera, Syrphidae) from the Kingdom of Bhutan, the easternmost representative of the bactrianus subgroup.

A new species of Eumerus, Eumerus druk Smit sp. nov., is described from Bhutan. This species belongs to the bactrianus subgroup of the strigatus species group. Seven species are currently known within this subgroup: four European, one of which is also found in the Near East, and three more only known from Tajikistan. The new species extends this disjunct distribution to the east by at least 2,000 km, stretching far beyond the reported Turano-Mediterranean region and into the Himalayas. A diagnosis and a key to all Central and Eastern Palaearctic species of the Eumerus bactrianus subgroup are provided.

A bacterial surface layer protein exploits multistep crystallization for rapid self-assembly.

Surface layers (S-layers) are crystalline protein coats surrounding microbial cells. S-layer proteins (SLPs) regulate their extracellular self-assembly by crystallizing when exposed to an environmental trigger. However, molecular mechanisms governing rapid protein crystallization in vivo or in vitro are largely unknown. Here, we demonstrate that the Caulobacter crescentus SLP readily crystallizes into sheets in vitro via a calcium-triggered multistep assembly pathway. This pathway involves 2 domains serving distinct functions in assembly. The C-terminal crystallization domain forms the physiological 2-dimensional (2D) crystal lattice, but full-length protein crystallizes multiple orders of magnitude faster due to the N-terminal nucleation domain. Observing crystallization using a time course of electron cryo-microscopy (Cryo-EM) imaging reveals a crystalline intermediate wherein N-terminal nucleation domains exhibit motional dynamics with respect to rigid lattice-forming crystallization domains. Dynamic flexibility between the 2 domains rationalizes efficient S-layer crystal nucleation on the curved cellular surface. Rate enhancement of protein crystallization by a discrete nucleation domain may enable engineering of kinetically controllable self-assembling 2D macromolecular nanomaterials.

New geographical records and key to the species of Eumerus Meigen, 1823 (Diptera, Syrphidae) introduced into the Americas and Hawaii

Abstract Eumerus Meigen, 1823 is a very speciose genus of flower flies from the Eastern Hemisphere. Several reports of introduced species of this genus in the Americas have been recorded since early in the twentieth century, with a present list of six species recorded to date from the Americas and the US territory of Hawaii. In this paper we give new geographical records for the African-native species Eumerus obliquus (Fabricius, 1805), which reflect the expansion of this fly through Brazil and Paraguay along the last twenty years. At the same time, we report a second species from Paraguay, Eumerus aurifrons (Wiedemann, 1824), being this the first Western Hemisphere record for this Asian-native species. We finally provide an identification key to all of the species of Eumerus presently known from the Americas and Hawaii.

Robber flies from Sint Eustatius, Lesser Antilles, with the descriptions of Efferia exaggerata sp. n. and the male of Ommatius prolongatus Scarbrough (Diptera: Asilidae).

Four species of robber flies are recorded from Sint Eustatius, Lesser Antilles. Efferia exaggerata sp. n. is described, as well as the male of Ommatius prolongatus Scarbrough.

Two new European long-legged hoverfly species of the Eumerusbinominatus species subgroup (Diptera, Syrphidae).

Eumerus Meigen (Diptera, Syrphidae) is one of the most speciose hoverfly genera in Europe, with several species groups recognized within. As part of the tricolor group of species, a subgroup of long-legged representatives stands out. We name it Eumerusbinominatus subgroup and provide descriptions for two new European species which belong to this subgroup: E.grallator sp. nov. from mainland Spain and E.tenuitarsis sp. nov. from Lesvos and Evros, Greece. A key for all five recognized species of the binominatus subgroup is provided.

A Caulobacter crescentus Microbicide Protects from Vaginal Infection with HIV-1JR-CSF in Humanized Bone Marrow-Liver-Thymus Mice.

Over 2 million people are infected with HIV-1 annually. Approximately half of these new infections occur in women residing in low-income countries, where their access to and control over HIV-1 preventative measures are often limited, indicating that female-controlled prevention options for HIV-1 are urgently needed. Microbicides that can be topically applied to the vaginal tract in advance of sexual activity represent a promising female-controlled prevention option for HIV-1. We have previously described the development of an HIV-1-specific microbicide using the surface or S-layer recombinant protein display capabilities of the nonpathogenic, freshwater bacterium Caulobacter crescentus Recombinant C. crescentus bacteria were created that displayed proteins that interfere with the HIV-1 attachment and entry process and that were able to provide significant protection of TZM-bl cells from infection with HIV-1 pseudovirus. These studies have been expanded to investigate if these recombinant C. crescentus bacteria are able to maintain efficacy with replication-competent HIV-1 and both TZM-bl cells and human peripheral blood mononuclear cells (PBMCs). In addition, we utilized the humanized bone marrow-liver-thymus (BLT) mouse model to determine if vaginal application of recombinant C. crescentus at the time of HIV-1JR-CSF infection could provide protection from HIV-1 infection. Recombinant C. crescentus bacteria expressing Griffithsin, GB virus C E2 protein, elafin, α-1-antitrypsin, indolicidin, and the fusion inhibitor T-1249 were able to protect 40 to 75% of the BLT mice from vaginal infection with HIV-1JR-CSF, with C. crescentus bacteria expressing Griffithsin being the most effective. Taken together, these data suggest that a C. crescentus-based microbicide could be a safe and effective method for HIV-1 prevention.IMPORTANCE Human immunodeficiency virus (HIV) disproportionally infects young women in sub-Saharan Africa. Current HIV-1 prevention options have had limited success among women, suggesting that alternative, female-controlled prevention options need to be developed. Microbicides that can be applied to the vaginal tract are a promising prevention option. In this study, we describe the testing of 15 potential candidates for inhibition of HIV-1 infection in a humanized mouse model of HIV-1 infection. Four of these candidates were able to provide significant protection from vaginal infection with HIV-1, with the most successful candidate protecting 75% of the mice from infection. This study describes the preclinical testing of a new strategy that could be a safe and effective option for HIV-1 prevention in women.

Topologically-guided continuous protein crystallization controls bacterial surface layer self-assembly.

Many bacteria and most archaea possess a crystalline protein surface layer (S-layer), which surrounds their growing and topologically complicated outer surface. Constructing a macromolecular structure of this scale generally requires localized enzymatic machinery, but a regulatory framework for S-layer assembly has not been identified. By labeling, superresolution imaging, and tracking the S-layer protein (SLP) from C. crescentus, we show that 2D protein self-assembly is sufficient to build and maintain the S-layer in living cells by efficient protein crystal nucleation and growth. We propose a model supported by single-molecule tracking whereby randomly secreted SLP monomers diffuse on the lipopolysaccharide (LPS) outer membrane until incorporated at the edges of growing 2D S-layer crystals. Surface topology creates crystal defects and boundaries, thereby guiding S-layer assembly. Unsupervised assembly poses challenges for therapeutics targeting S-layers. However, protein crystallization as an evolutionary driver rationalizes S-layer diversity and raises the potential for biologically inspired self-assembling macromolecular nanomaterials.

Bat Ectoparasites From Sint Eustatius, Lesser Antilles (Diptera: Hippoboscidae: Streblinae; Hemiptera: Polyctenidae).

In this paper 4 species of bat ectoparasites are recorded from the island of Sint Eustatius, Dutch Caribbean. One species of true bug (Hemiptera: Polyctenidae) as well as 3 species of bat flies (Diptera: Hippoboscidae: Streblinae) are recorded. All species are photographed. The first DNA barcodes for 3 bat ectoparasite species ( Trichobius frequens, Trichobius intermedius, and Hesperoctenes fumarius) have been posted to the BOLD database; DNA barcode sequences for a fourth species ( Megistopoda aranea) are the first from a Caribbean island.

Tolmerus calceatus (Meigen) confirmed as a valid species separate from Tolmerus atricapillus (Fallén) (Diptera: Asilidae).

The status of Tolmerus atricapillus calceatus (Meigen) is discussed. Based on morphological, ecological as well as genetic information it is concluded that Tolmerus calceatus stat. rev. should be treated as a valid species separate from T. atricapillus (Fallén). Characters to separate both are provided as well as a key to all Tolmerus species of northwestern Europe.

First host record of Myopa pellucida Robineau-Desvoidy (Diptera: Conopidae) identified using DNA barcoding.

Thick-headed flies of the genus Myopa Fabricius are a common sight in early spring in Europe. Several species can be found flying among Salix catkins together with their supposed Andrena Fabricius hosts (Hymenoptera: Apidae) (John Smit observations). Despite the fact that some of these Myopa species are very common, little is known about their actual host-parasitoid associations. Only a few scattered host records can be found in the literature, most of which refer to M. testacea s.l.. Whether or not these records actually concern M. testacea (Linnaeus) is uncertain due to the confusion over the identity of several species within the M. testacea species group (Stuke Clements 2008). Stuke (2017), in his World Catalogue of Conopids, provides all recorded host associations and distinguishes between unambiguously accepted host records and doubtful records based on three criteria: 1) the conopid has been reared from the host or a larva has been identified by DNA barcoding; 2) there is no doubt concerning the identity of the conopid species concerned, and 3) there is no doubt concerning the identity of the host species. Because the vast majority of the host records in the literature do not fulfil one or more of these criteria they are regarded as doubtful (Stuke 2017). Thus, only three host associations for the genus Myopa can be regarded as confirmed: Andrena vaga Panzer, for both M. hirsuta Stuke Clements (Jentzsch 2009) and M. testacea (Erteld 1998, Fellendorf et al. 2004, De Meijere 1912), and the Nearctic Andrena regularis Malloch, for the Holarctic M. vicaria Walker (Miliczky Osgood 1995). Here we report on a fourth host association for the genus Myopa, since a second instar larva of M. pellucida Robineau-Desvoidy was found in the abdomen of an Andrena nitida (Müller) female and identified using DNA barcoding.

Generation of a Dual-Target, Safe, Inexpensive Microbicide that Protects Against HIV-1 and HSV-2 Disease.

HSV-2 infection is a significant health problem and a major co-morbidity factor for HIV-1 acquisition, increasing risk of infection 2-4 fold. Condom based prevention strategies for HSV-2 and HIV-1 have not been effective at stopping the HIV-1 pandemic, indicating that alternative prevention strategies need to be investigated. We have previously developed an inexpensive HIV-1 specific microbicide that utilizes the S-layer mediated display capabilities of Caulobacter crescentus, and have shown that recombinant C. crescentus displaying HIV entry blocking proteins are able to provide significant protection from HIV-1 infection in vitro. Here we demonstrate that recombinant C. crescentus are safe for topical application and describe 5 new recombinant C. crescentus that provide protection from HIV-1 infection in vitro. Further, we demonstrate protection from disease following intravaginal infection with HSV-2 in a murine model using C. crescentus expressing the anti-viral lectins Cyanovirin-N and Griffithsin, as well as α-1-antitrypsin and indolicidin. Interestingly, C. crescentus alone significantly reduced HSV-2 replication in vaginal lavage fluid. Protection from HSV-2 disease was strongly associated with early cytokine production in the vaginal tract. Our data support the potential for a dual-target microbicide that can protect against both HIV-1 and HSV-2, which could have an enormous impact on public health.

Proposal of Henriciella barbarensis sp. nov. and Henriciella algicola sp. nov., stalked species of the genus and emendation of the genus Henriciella.

Two Gram-negative, heterotrophic, aerobic, prosthecated, marine bacteria, designated strains MCS23T and MCS27T, were isolated from seawater samples. NaCl was required for growth. The major polar lipid detected in strain MCS27T was phosphatidylglycerol, whereas those detected in MCS23T were phosphatidylglycerol, sulfoquinovosyl diacylglycerol and 1,2-diacyl-3-α-d-glucuronopyranosyl-sn-glycerol taurineamide. The most abundant cellular fatty acids were C18 : 1ω7 and C16 : 0, hydroxyl-fatty acids were 3-OH C12 : 0 in both strains and 3-OH C11 : 0 in MCS23T. Strains MCS23T and MCS27T had DNA G+C contents of 57.0 and 55.0 mol%, respectively. The two strains shared 99.3 % 16S rRNA gene sequence similarity; levels of similarity with the type strains of species of the genus Henriciella were 99.4-97.8 % but DNA-DNA hybridizations were 53 % or lower. Besides their 16S rRNA gene sequences, the novel strains can be differentiated from other species of the genus Henriciella by cell morphology, lipid and fatty acid patterns and enzyme activities. The data obtained led to the identification of two novel species, for which the names Henriciella barbarensis sp. nov. (type strain MCS23T=LMG 28705T=CCUG 66934T) and Henriciella algicola sp. nov. (type strain MCS27T=LMG 29152T=CCUG 67844T) are proposed. As these two novel species are the first prosthecate species in the genus Henriciella, an emended genus description is also provided.

Environmental Calcium Controls Alternate Physical States of the Caulobacter Surface Layer.

Surface layers (S-layers) are paracrystalline, proteinaceous structures found in most archaea and many bacteria. Often the outermost cell envelope component, S-layers serve diverse functions including aiding pathogenicity and protecting against predators. We report that the S-layer of Caulobacter crescentus exhibits calcium-mediated structural plasticity, switching irreversibly between an amorphous aggregate state and the crystalline state. This finding invalidates the common assumption that S-layers serve only as static wall-like structures. In vitro, the Caulobacter S-layer protein, RsaA, enters the aggregate state at physiological temperatures and low divalent calcium ion concentrations. At higher concentrations, calcium ions stabilize monomeric RsaA, which can then transition to the two-dimensional crystalline state. Caulobacter requires micromolar concentrations of calcium for normal growth and development. Without an S-layer, Caulobacter is even more sensitive to changes in environmental calcium concentration. Therefore, this structurally dynamic S-layer responds to environmental conditions as an ion sensor and protects Caulobacter from calcium deficiency stress, a unique mechanism of bacterial adaptation. These findings provide a biochemical and physiological basis for RsaA's calcium-binding behavior, which extends far beyond calcium's commonly accepted role in aiding S-layer biogenesis or oligomerization and demonstrates a connection to cellular fitness.