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To probe its environment, the flying insect controllably flexes, twists, and maneuvers its antennae by coupling mechanical deformations with the sensory output. We question how the materials properties of insect antennae could influence their performance. A comparative study was conducted on four hawkmoth species: Manduca sexta, Ceratomia catalpae, Manduca quinquemaculata , and Xylophanes tersa. The morphology of the antennae of three hawkmoths that hover while feeding and one putatively non-nectar-feeding hawkmoth (Ceratomia catalpa) do not fundamentally differ, and all the antennae are comb-like (i.e., pectinate), markedly in males but weakly in females. Applying different weights to the free end of extracted cantilevered antennae, we discovered anisotropy in flexural rigidity when the antenna is forced to bend dorsally versus ventrally. The flexural rigidity of male antennae was less than that of females. Compared with the hawkmoths that hover while feeding, Ceratomia catalpae has almost two orders of magnitude lower flexural rigidity. Tensile tests showed that the stiffness of male and female antennae is almost the same. Therefore, the differences in flexural rigidity are explained by the distinct shapes of the antennal pectination. Like bristles in a comb, the pectinations provide extra rigidity to the antenna. We discuss the biological implications of these discoveries in relation to the flight habits of hawkmoths. Flexural anisotropy of antennae is expected in other groups of insects, but the targeted outcome may differ. Our work offers promising new applications of shaped fibers as mechanical sensors. Insect antennae are blood-filled, segmented fibers with muscles in the two basal segments. The long terminal segment is muscle-free but can be flexed. Our comparative analysis of mechanical properties of hawkmoth antennae revealed a new feature: antenna resistance to bending depends on the bending direction. Our discovery replaces the conventional textbook scenario considering hawkmoth antennae as rigid rods. We showed that the pectinate antennae of hawkmoths behave as a comb in which the bristles resist bending when they come together. This anisotropy of flexural resistance offers a new mode of environmental sensing that has never been explored. The principles we found apply to other insects with non-axisymmetric antennae. Our work offers new applications for shaped fibers that could be designed to sense the flows. [Display omitted] [ABSTRACT FROM AUTHOR]

Insect antennae are hollow, blood-filled fibers with complex shape. Muscles in the two basal segments control antennal movement, but the rest (flagellum) is muscle-free. The insect can controllably flex, twist, and maneuver its antennae laterally. To explain this behavior, we performed a comparative study of structural and tensile properties of the antennae of Periplaneta americana (American cockroach), Manduca sexta (Carolina hawkmoth), and Vanessa cardui (painted lady butterfly). These antennae demonstrate a range of distinguishable tensile properties, responding either as brittle or strain-adaptive fibers that stiffen when stretched. Scanning electron microscopy and high-speed imaging of antennal breakup during stretching revealed complex coupling of blood pressure and cuticle deformation in antennae. A generalized Lamé theory of solid mechanics was developed to include the force-driven deformation of blood-filled antennal tubes. We validated the theory against experiments with artificial antennae with no adjustable parameters. Blood pressure increased when the insect inflated its antennae or decreased below ambient pressure when an external tensile load was applied to the antenna. The pressure–cuticle coupling can be controlled through changes of the blood volume in the antennal lumen. In insects that do not fill the antennal lumen with blood, this blood pressure control is lacking, and the antennae react only by muscular activation. We suggest that the principles we have discovered for insect antennae apply to other appendages that share a leg-derived ancestry. Our work offers promising new applications for multifunctional fiber-based microfluidics that could transport fluids and be manipulated by the same fluid on demand. Insect antennae are blood-filled, segmented fibers with muscles in the two basal segments. The long terminal segment is muscle-free but can be flexed. To explain this behavior, we examined structure-function relationships of antennae of cockroaches , hawkmoths, and butterflies. Hawkmoth antennae behaved as brittle fibers, but butterfly and cockroach antennae showed strain-adaptive behavior like fibers that stiffen when stretched. Videomicroscopy of antennal breakup during stretching revealed complex coupling of blood pressure and cuticle deformation. Our solid mechanics model explains this behavior. Because antennae are leg-derived appendages, we suggest that the principles we found apply to other appendages of leg-derived ancestry. Our work offers new applications for multifunctional fiber-based microfluidics that could transport fluids and be manipulated by the fluid on demand. [Display omitted] [ABSTRACT FROM AUTHOR]

Abstract A recently described species of black fly, Simulium wayani Takaoka and Chen, from the island of Timor was chromosomally mapped to provide insights into its evolutionary and biogeographic history. The morphologically based species status of S. wayani is supported by a suite of fixed chromosomal rearrangements and unique sex chromosomes derived primarily from a large pool of polymorphisms in the S. ornatipes complex in Australia. The banding patterns of its polytene chromosomes indicate that S. wayani is closely related to a pair of homosequential cryptic species (S. norfolkense Dumbleton and S. ornatipes cytoform A2) in the S. ornatipes Skuse complex on mainland Australia; all three species uniquely share the same amplified band in their chromosomal complement. The low level of polymorphism and heterozygosity in S. wayani , relative to Australian populations of the S. ornatipes complex, suggests few colonization events from the larger land mass. [ABSTRACT FROM AUTHOR]

The polytene chromosomes of 130 larvae of the Neotropical Simulium metallicum complex from Brazil, Costa Rica, and Ecuador revealed five cytoforms, including three (‘M’, ‘N’, and ‘O’) that are new and two (‘B’ and ‘J’) that represent range extensions of up to 850 km. The discovery of three new cytoforms brings the total number in the complex to 17. Cytoforms ‘B’, ‘J’, and ‘N’ are reproductively isolated from one another, and their species status is corroborated by morphological evidence. None of the three new cytoforms is known from current or historical onchocerciasis foci, although ‘M’ inhabits the periphery of the former Ecuadorian Santiago onchocerciasis focus a mere 30 km to the west. The number of fixed chromosomal differences, as many as 24, separating some members of the S. metallicum complex far exceeds that known between members of any other simuliid species complex. Two distinct groupings can be diagnosed within the S. metallicum complex, based on at least eight fixed chromosomal rearrangements and structural characters in the larval stage. Consequently, a recommendation is made to recognize the S. horacioi complex and the S. metallicum complex sensu stricto . Recognition of two separate complexes provides potential phylogenetic content with predictive power for understanding biological phenomena such as vector potential. [ABSTRACT FROM AUTHOR]

Studies of the lowland avifauna in the Neotropical Region have shown a paucity of Leucocytozoon species. However, surveys conducted in the Colombian highlands revealed a great diversity of these parasites infecting resident birds. To further investigate the relationship between Leucocytozoon diversity, the potential vectors, and altitudinal distribution, birds from 41 families were sampled at low and high elevations in Colombia. Blood samples were screened by microscopy, and a fragment of cytochrome b was amplified from Leucocytozoon -positive samples. The complete mitochondrial genome was also obtained for each morphospecies of Leucocytozoon . Leucocytozoon species were detected in resident birds, with various degrees of host specificity, at elevations from 2,400 to 3,950 meters above sea level, where five new host-parasite associations were discovered. Phylogenetic analysis based on the cytochrome b fragment suggested that two nominal taxa, L. fringillinarum and L. majoris , are species complexes. Blood sources of Simuliidae revealed generalist-feeding habits that included avian and mammalian hosts. Molecular analysis of parasites in black flies indicated a close relationship with the parasites found in birds. Our investigation provides further evidence that the distribution and transmission of Leucocytozoon species in the Neotropics are influenced by elevation, with the highest prevalence between 2,400 and 3,200 m asl. [ABSTRACT FROM AUTHOR]

Abstract: More than 2000 species of black flies feed on vertebrate blood; 1.5% of all species are vectors of pathogens that cause human diseases. Of nine simuliid-borne animal diseases, only two, mansonellosis and onchocerciasis, afflict humans. Onchocerciasis is a debilitating disease infecting an estimated 40 million people in Africa, Latin America, and Yemen, whereas mansonellosis is a mild disease in the Neotropics. Cytogenetic studies of natural populations of more than 500 species of black flies have revealed that the classic morphospecies of taxonomists is typically a complex of two or more reproductively isolated entities, or sibling (cryptic) species. Most vectors of human pathogens are sibling species, each ecologically unique in traits such as breeding habitats, dispersal capabilities, and degree of vector competence. We review the evolution of black flies, the cytogenetics that have revealed about 260 cytologically distinct entities, the molecular studies that continue to expose additional hidden biodiversity, and a case study of the epidemiology of the Simulium damnosum complex, the largest species complex of blood-feeding arthropods on Earth and the premier group of black flies responsible for human onchocerciasis. [Copyright &y& Elsevier]

Study objective: Previous studies have suggested that QTc prolongation may lead to significant morbidity and mortality. The prevalence of QTc prolongation among emergency department (ED) patients is unknown. The purpose of this study is to determine the prevalence of QTc prolongation among ED patients. Methods: This was a retrospective review of ED and inpatient data for all patients with an ECG conducted for any reason at a tertiary care university ED during a 3-month period. QTc prolongation was defined as computer-generated QTc intervals greater than or equal to 450 ms for men and greater than or equal to 460 ms for women. Results: Of the 1,558 eligible cases, 544 patients had QTc prolongation (35%; 95% confidence interval [CI] 32% to 37%). The prevalence of QTc intervals greater than or equal to 500 ms was 8% (120/1,558; 95% CI 6% to 9%). The most common comorbidities were structural heart disease, renal failure, and stroke. Forty-four percent (239/544; 95% CI 40% to 48%) of patients with any degree of QTc prolongation were discharged from the ED. Furthermore, 23% (28/120; 95% CI 16% to 32%) of patients with QTc intervals greater than or equal to 500 ms were discharged from the ED, including 16 patients with QTc intervals greater than or equal to 500 ms and QRS durations less than 120 ms (16/60; 27%; 95% CI 16% to 40%). Five percent of the patients with QTc prolongation died in the ED or during hospitalization (27/544; 95% CI 3% to 7%); none had QTc prolongation or torsades de pointes listed as a cause of death. Conclusion: QTc prolongation occurred frequently among ED patients who had an ECG study for any reason. Nearly half of all patients with QTc prolongation were discharged from the ED. [Copyright &y& Elsevier]

• Black flies in the Simulium vernum group reach their southern limit in Southeast Asian mountains. • Chromosomes demonstrate reproductive isolation of Simulium chomthongense and S. khunklangense. • Polytene chromosomes reveal the existence of cryptic species (A and B) of S. chomthongense. • All three species are endemic or nearly endemic near the top of Doi Inthanon. • Possible sympatric speciation and conservation of high-elevation streams are suggested. High-mountain summits in tropical regions are habitat islands that provide refugia for northern taxa and opportunities for speciation. These focal hotspots of biodiversity are also threatened by increasing anthropogenic pressures and changing climatic conditions. The largest species group of black flies, the Simulium vernum group, reaches its southern limit in the mountains of Southeast Asia, providing an opportunity to examine macrogenetic diversification at the southern edge of the group's range. We investigated the polytene chromosomes of two morphologically similar species in the S. vernum group, S. chomthongense Takaoka, Srisuka & Choochote, and S. khunklangense Takaoka & Srisuka, from their type localities near the summit of Doi Inthanon, Thailand's highest mountain. Chromosomal analyses of 112 larvae demonstrated reproductive isolation between S. chomthongense and S. khunklangense and revealed a cryptic species of S. chomthongense. The three species have the same fixed banding sequence in five of the six chromosome arms, but they are distinguished by the successive addition of overlapping inversions in the IIIL arm. The two cytospecies (A and B) of S. chomthongense s. l. are the third example of endemic or nearly endemic cryptic species pairs near the top of Doi Inthanon, suggesting the possibility of sympatric speciation and the importance of conserving high-elevation stream habitats. Cytogenetic analysis of black flies in the Simulium vernum group revealed cryptic species and demonstrated reproductive isolation among three species near the top of Thailand's highest mountain, Doi Inthanon. Image, graphical abstract [ABSTRACT FROM AUTHOR]

Simulium perplexum Shelley, Maia-Herzog, Luna Dias & Couch is structurally similar in the pupal stage to Simulium guianense Wise, the main vector in the onchocerciasis foci of Amazonian Brazil and Venezuela. We report S. perplexum for the first time beyond its type locality (Guyana, Potaro River), describe its larva, redescribe its pupa, and provide a chromosomal comparison with S. guianense and other morphologically similar species. We collected it in two rivers in Rurópolis municipality, Pará state, Brazil. The larvae can be distinguished from those of related species by having body cuticle with microscopic, translucent, and lanceolate setae. Chromosomal comparisons of S. perplexum and similar Brazilian species with available chromosome information ( S. guianense , S. hirtipupa Lutz, and S. litobranchium Hamada, Pepinelli, Mattos-Glória & Luz), using S. guianense Cytoform A as the standard, show that S. perplexum has the nucleolar organizer uniquely in the middle of the short arm of chromosome I, whereas the other three species have this marker at the base of the long arm of chromosome I. All chromosome arms, except IIS and IIIS, of S. perplexum are rearranged, compared with S. guianense Cytoform A, suggesting that it is not closely related to this species or to S. litobranchium , as suggested by some authors, based on morphological features. [ABSTRACT FROM AUTHOR]

Abstract: The Blastocladiomycota is a recently described phylum of ecologically diverse zoosporic fungi whose species have not been thoroughly sampled and placed within a molecular phylogeny. In this study, we investigated the phylogeny of the Blastocladiomycota based on ribosomal DNA sequences from strains identified by traditional morphological and ultrastructural characters. Our results support the monophyly of the Coelomomycetaceae and Physodermataceae but the Blastocladiaceae and Catenariaceae are paraphyletic or polyphyletic. The data support two clades within Allomyces with strains identified as Allomyces arbusculus in both clades, suggesting that species concepts in Allomyces are in need of revision. A clade of Catenaria species isolated from midge larvae group separately from other Catenaria species, suggesting that this genus may need revision. In the Physodermataceae, Urophlyctis species cluster with a clade of Physoderma species. The algal parasite Paraphysoderma sedebokerensis nom. prov. clusters sister to other taxa in the Physodermataceae. Catenomyces persicinus, which has been classified in the Catenariaceae, groups with the Chytridiomycota rather than Blastocladiomycota. The rDNA operon seems to be suitable for classification within the Blastocladiomycota and distinguishes among genera; however, this region alone is not suitable to determine the position of the Blastocladiomycota among other basal fungal phyla with statistical support. A focused effort to find and isolate, or directly amplify DNA from additional taxa will be necessary to evaluate diversity in this phylum. We provide this rDNA phylogeny as a preliminary framework to guide further taxon and gene sampling and to facilitate future ecological, morphological, and systematic studies. [Copyright &y& Elsevier]

• Cytoform 'L' of S. tani in Taiwan, is described as a new species S. jianshiense • This new species is morphologically and genetically separated from the S. suzukii complex cytoforms 'C' and 'D' • S. ryukyuense is revalidated and applied to cytoform 'D' The species status of two cytoforms of two species complexes in the Simulium (Simulium) tuberosum species-group, which are almost indistinguishable in the pupal stage, was morphologically and genetically evaluated. Cytoform 'L' of the S. (S.) tani Takaoka & Davies complex, previously recognized as S. (S.) suzukii Rubtsov in Taiwan, is described as a new species, S. (S.) jianshiense. It is morphologically distinguishable as adults from two members of the S. (S.) suzukii complex: cytoform 'C' from Hokkaido, selected to represent the type of S. (S.) suzukii sensu stricto, and cytoform 'D' from Okinawa Island and, based on our chromosomal analysis, also from Amami Island. This new species is genetically separated from both cytoforms of the S. (S.) suzukii complex with a genetic distance of 5.31–6.67%. Cytoform 'D' is distinguished from cytoform 'C' by the color of the male forecoxa and relative length of the female sensory vesicle to the third maxillary palpomere. Accordingly, the name S. (S.) ryukyuense Ogata, once regarded as a synonym of S. (S.) suzukii , is revalidated and applied to cytoform 'D'. The genetic distance between S. (S.) ryukyuense and S. (S.) suzukii sensu stricto is 1.24–1.60%. [ABSTRACT FROM AUTHOR]

• First culturable bacteria from one of Malaysia's most common black flies, Simulium tani were investigated using 16S rDNA sequencing • 20 isolates representing 10 bacterial species in four genera were found. • Five isolates showed β-hemolysis on Columbia agar, and virulence genes were found. • Some degree of resistance was found among the isolates. • The study suggests rich opportunities for comparative studies exploring the diversity and roles of the microbiome of S. tani and other Southeast Asian black flies. Although the microbiome of blood-feeding insects serves an integral role in host physiology, both beneficial and pathogenic, little is known of the microbial community of black flies. An investigation, therefore, was undertaken to identify culturable bacteria from one of Malaysia's most common black flies, Simulium tani Takaoka and Davies, using 16S rDNA sequencing, and then evaluate the isolates for antibiotic resistance and virulence genes. A total of 20 isolates representing 11 bacterial species in four genera were found. Five isolates showed β-hemolysis on Columbia agar, and virulence genes were found in three of these isolates. Some degree of resistance to six of the 12 tested antibiotics was found among the isolates. The baseline data from this study suggest rich opportunities for comparative studies exploring the diversity and roles of the microbiome of S. tani and other Southeast Asian black flies. [ABSTRACT FROM AUTHOR]

• The coding sequences of serine proteases of Hypoderma bovis were characterized. • A phylogeny of HA, HB, and HC genes among Hypoderma species was inferred. • rHC was expressed and purified by affinity chromatography as a diagnostic antigen. • rHC and a newly designed sHC were evaluated in iELISA for bovine hypodermosis. • rHC and sHC iELISAs had sensitivities higher than 90 % and specificities of 100 %. Hypodermins A (HA), B (HB), and C (HC) of warble flies are modulatory antigens involved in host inflammation and immune responses during migration of the warble fly larvae through host connective tissues. In the current study, molecular characteristics of the genes encoding HA, HB, and HC were revealed from cDNA constructs of third-instar larvae of Hypoderma bovis. The open reading frame (ORF) of each hypodermin gene was amplified with modified gene-specific primers, and the resulting PCR products were cloned into pGEM-T Easy Vector to produce recombinant plasmids (rHA, rHB, and rHC). The ORF sequences of rHA, rHB, and rHC genes are 705 bp, 771 bp, and 783 bp long and encode proteins of 234, 256, and 263 amino acids with predicted sizes of 25.74 kDa, 27.79 kDa, and 28.51 kDa, respectively. The rHC gene was subcloned into the pET 100/D-TOPO Expression Vector, and the recombinant HC was purified using affinity chromatography. Western blotting indicated that rHC was recognized by the sera of cattle naturally infested with H. bovis. The rHC and a synthetic peptide (sHC) containing its linear B cell-specific epitope were evaluated as serological markers in indirect ELISA (iELISA) for the diagnosis of bovine hypodermosis. Both sHC and rHC iELISAs had sensitivity values equal to or higher than 90 % and specificity values of 100 %. A total of 200 serum samples from cattle in the Central Anatolia Region of Turkey were also analyzed by rHC and sHC-iELISAs to reveal the seroprevalence of bovine hypodermosis. The results of both iELISAs were consistent with one another and revealed a hypodermosis prevalence of 62 %. Our study provides the first data on molecular characterization of hypodermin genes of H. bovis and indicates the efficacy of recombinant antigen and peptide-based iELISA for serodiagnosis of bovine hypodermosis. [ABSTRACT FROM AUTHOR]

• Biting of humans by simuliids, and simuliotoxicosis in cattle, is common in Aras River Basin, Iran. • Twelve simuliid morphotaxa were captured in the Aras River Basin. • Simulium turgaicum complex, with 97% relative abundance, was the dominant taxon. • Three molecular lineages in the S. turgaicum complex were found. • High abundance and biting of humans by S. turgaicum are important epidemiological factors. Black flies are insects of medical, veterinary, and environmental significance. Historically, they have attacked humans and caused simuliotoxicosis in livestock in the Aras River Basin in northwest Iran. However, information on the species and their bionomics is limited in the region. Adult flies were collected from diverse ecotopes of the Aras River Basin. After morphological identification, representative specimens of each morphological group were subjected to mtDNA COI gene sequence analysis for species diagnosis and to infer relationships. Flies also were examined for pollinia. A total of 1880 specimens representing 12 morphotaxa in two genera (Simulium and Metacnephia) were identified: Simulium turgaicum (n=1834), S. kiritshenkoi (n=12), S. bezzii (n=7), S. brevitarse (n=7), S. pseudequinum (n=5), S. aureum species group (n=4), S. vernum species group (n=3), S. transcaspicum (n=1), three unidentified species of the subgenus Simulium (n=5), and Metacnephia possibly persica (n=2). Fifty two haplotypes were detected for the 65 COI sequences analyzed. Intraspecific genetic divergence was 0.19-8.83%, whereas the mean interspecific genetic distances among the morphotaxa were 1.41-19.58%. Molecular analyses recovered three well-supported lineages within S. turgaicum. One lineage included black flies collected from agricultural fields, a second lineage involved black flies captured from animals, and a third lineage included specimens that had visited flowers, as evidenced by presence of pollinia. The relative abundance (97%) and observations of the S. turgaicum complex biting humans are important epidemiological factors. Future studies are needed to define the potential epidemiological risk of simulids in Khoda-Afarin County of Iran. Image, graphical abstract [ABSTRACT FROM AUTHOR]

The ecology of black flies in Thailand was investigated, based on 19,451 larvae and pupae collected from 65 stream sites in 10 northern provinces during the rainy, cool, and hot seasons, and 1,906 larvae and pupae collected from 18 sites in 9 southern provinces during the cool season. Twenty-seven black fly species were identified from northern Thailand, of which 26 were found in the cool season, when richness was greatest. Significant regressions between species richness and elevation fit a unimodal model in the rainy season but a linear model in the cool and hot seasons. Twenty-two species occurred in all seasons. Species in the subgenera Gomphostilbia and Nevermannia were most common in the hot season, whereas species in the subgenus Simulium were predominant in the cool season. Some species (e.g., S. nakhonense) were geographically widespread, whereas others (e.g., S. chaliowae and S. weji) were restricted to particular localities. Eighteen species and species complexes were found in southern Thailand. The S. tani complex was the most widely distributed taxon, occurring at 66.7% of the sites in the South. Ecological analyses revealed that water temperature, elevation, conductivity, dissolved oxygen, and stream size were among the significant factors associated with the distributions of black flies in both regions of Thailand—the same factors associated with simuliid distributions in other areas of the world. [ABSTRACT FROM AUTHOR]

• Simuliid pest problems and simuliotoxicosis affect cattle in northeastern Turkey. • The COI gene was analyzed for four species of black flies in the pest region. • Genetic divergence was 0.2–1.6% (intraspecific) and 11.2–14.5% (mean interspecific). • COI results reflect nominal species but do not unequivocally reveal cryptic species. • A rapid identification tool now exists to aid simuliid control in Turkey's pest area. Accurate species identification provides the foundation for successful pest management and vector control of black flies. Accordingly, we examined the mitochondrial DNA cytochrome oxidase I (COI) gene sequences of four morphologically and chromosomally identified species of black flies (Simulium vernum group sp., S. bergi Rubtsov, S. bezzii (Corti), and S. kiritshenkoi Rubtsov) in Northeast Anatolia Region of Turkey where simuliid pest problems and simuliotoxicosis cases have been reported among cattle. COI gene sequences of these species and closely related species available in GenBank were used to provide species-level diagnoses and infer relationships. Both subgenera (Nevermannia and Simulium) were monophyletic, and subclades generally corresponded with species groups. Intraspecific genetic divergence was 0.2–1.6%, whereas the mean interspecific genetic divergence among the four species was 11.2–14.5%. The COI analysis produced results congruent with morphological concepts of the nominal species S. bergi and S. bezzii. Probable misidentifications in GenBank were revealed, especially for species in the S. ornatum and S. vernum groups, complicating identification capability. Sequence variation in the COI barcode region also might not be adequate for species delineation and identification among some species in these two species groups. [ABSTRACT FROM AUTHOR]