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ABSTRACT The present study provides a comprehensive overview of the Megaloptera fauna in Brazil. A total of 27 species-25 extant and two extinct-distributed into two families, Corydalidae and Sialidae, and six genera, are recorded from Brazil. The historical timeline of Megalopteran records in Brazil spans 180 years, from 1842 to 2022, and unfolds into three distinct periods: the 19th century and the first half of the 20th century dominated by European authors, the second half of the 20th century dominated by Mexican and US-born authors, and the 21st century dominated by Brazilian authors. Currently, about 75% of the type specimens of Brazilian Megaloptera are housed in foreign institutions. Among Brazilian states, Minas Gerais boasts the highest number of megalopteran records, with eight, while eight states have no records. The biome of Atlantic Forest exhibits the greatest richness of Megaloptera, with 15 species, whereas Pampas and Pantanal lack records of the order. The Amazon Basin leads with ten recorded species, although three Brazilian hydrographic basins remain with no records. Impressively, only ten Brazilian Megaloptera species have known immature stages.

Eggs of insects are immobile and must endure harsh environmental conditions (e.g., low temperatures in winter and aridity in summer) and avoid attack by egg-eating predators, egg parasites, and microbes. Females of Megaloptera lay their eggs as a single- or multi-layered egg mass, which is coated with chemical substances secreted from the female reproductive accessory glands. In this study, we observed the egg masses laid by females of two species of Sialidae (alderflies), nine species of Chauliodinae (fishflies), and 23 species of Corydalinae (dobsonflies) belonging to the order Megaloptera and examined the functions of accessory gland substances coating the laid eggs. The female accessory gland is a single tube in alderflies and fishflies but a paired pouch in dobsonflies. The amount and color of the gland substances differ greatly among species. These substances prevent egg desiccation, inhibit egg feeding by ladybird beetles, and repel ants. Most characteristics of the egg mass structures and the effectiveness of accessory gland substances reflect the phylogeny of Megaloptera, although some differ among closely related taxa.

In urban river systems, fragmentation of habitats and in-stream dispersal barriers play a major role in shaping the population genetic structure of freshwater macroinvertebrate species. In small, fragmented populations, effects of genetic drift and inbreeding are enhanced, which can lead to increased population differentiation and genetic diversity loss. One formerly strongly degraded and fragmented stream system in a highly urbanised area is the Emscher catchment in North Rhine-Westphalia, Germany. Major restoration efforts have led to an improvement of water and habitat quality over the past 20 years also in the formerly polluted tributaries, for example, the Boye catchment. However, the analysis of the population structure of two different amphipod and isopod species has revealed that some populations are still strongly isolated, indicating persisting gene flow barriers. In contrast, the effects are expected to be less pronounced in merolimnic species, which have an adult winged life stage, such as the alderfly (Linnaeus, 1758) . However, this species was much less abundant in the Boye catchment and not found in adjacent catchments (only 9 of 41 sampling sites), reducing the power of possible analyses. As no population genetic studies of have to our knowledge been published so far and genetic resources are generally scarce for this species, we generated and present here population genetic data for 70 specimens for the mitochondrial cytochrome c oxidase I (COI) gene and, more importantly, high resolution genomic single nucleotide polymorphism (SNP) data for 71 specimens, generated with double-digest restriction site-associated sequencing (ddRAD-seq). These data can be valuable for further studies, analysing the population genetic structure, dispersal pathways and potential gene flow barriers for on a larger geographic scale. Additional to presenting the data, we also give first insights in the population structure on a small geographic scale (area of approx. 15 km ). While the population differentiation was generally low, as expected on this small scale, we still found that gene flow was not equally strong between all populations, but that one population played a central role as a source and sink population, which cannot only be explained by the distance between populations.

Megaloptera are a basal holometabolous insect order with larvae exclusively predacious and aquatic. The evolutionary history of Megaloptera attracts great interest because of its antiquity and important systematic status in Holometabola. However, due to the difficulties identifying morphological apomorphies for the group, controversial hypotheses on the monophyly and higher phylogeny of Megaloptera have been proposed. Herein, we describe the complete mitochondrial (mt) genome of a fishfly species, Neochauliodes punctatolosus Liu & Yang, 2006, representing the first mt genome of the subfamily Chauliodinae. A phylogenomic analysis was carried out based on the mt genomic sequences of 13 mt protein-coding genes (PCGs) and two rRNA genes of nine Neuropterida species, comprising all three orders of Neuropterida and all families and subfamilies of Megaloptera. Both maximum likelihood and Bayesian inference analyses highly support the monophyly of Megaloptera, which was recovered as the sister of Neuroptera. Within Megaloptera, the sister relationship between Corydalinae and Chauliodinae was corroborated. The divergence time estimation suggests that stem lineage of Neuropterida and Coleoptera separated in the Early Permian. The interordinal divergence within Neuropterida might have occurred in the Late Permian.

Chemical secretions are an effective means by which insects may deter potential enemies, already being studied extensively with regard to their chemicals, synthesis, toxicity, and functions. However, these defensive secretions have been little studied in Megaloptera. Herein, the fluid expelling behavior of adult alderflies (Sialidae), fishflies (Corydalidae: Chauliodinae), and dobsonflies (Corydalidae: Corydalinae), all of the order Megaloptera, is described in detail regarding the timing and possible function of this behavior. When disturbed artificially, both males and females could expel fluid from the anus. However, the frequency of expelling was much lower in alderflies than in fishflies and dobsonflies. The amount of expelled fluid relative to body weight was also smaller in alderflies. In fishflies and dobsonflies, the amount of expelled fluid decreased with adult age, probably because the fluid is little replenished once expelled. The cream-colored fluid seems to be meconial fluid produced via the Malpighian tubules at the pupal stage, which is usually discharged at adult emergence in most other holometabolous insects. However, adult fishflies and dobsonflies often expel it vigorously by bending their abdomen when disturbed after emergence. Thus, the fluid expelling may be an anti-predatory behavior, particularly in younger adults that can expel a relatively large amount of fluid.

Megaloptera belong to a large monophyletic group, the Neuropteroidea, together with Coleoptera, Strepsiptera, Raphidioptera, and Neuroptera. With the latter two, this order constitutes the Neuropterida, a smaller monophyletic subset among which it is the only entirely aquatic group, with larvae of all species requiring submersion in freshwater. Megaloptera is arguably the oldest extant clade of Holometabola with aquatic representatives, having originated during the Permian before the fragmentation of Pangea, since about 230 Ma. It includes 54 genera (35 extant and 19 extinct genera), with 397 extant described species and subspecies. Recent Megaloptera are divided into two families: Corydalidae (with subfamilies Corydalinae—dobsonflies and Chauliodinae—fishflies) and Sialidae (alderflies), both widely yet disjunctively distributed among zoogeographical realms. All species of Megaloptera have aquatic larvae, whereas eggs, pupae, and adults are terrestrial. The anatomy, physiology, and behavior of megalopteran larvae are specialized for an aquatic predatory habit, yet their ecological significance might still be underappreciated, as their role in food webs of benthic communities of many temperate and tropical streams and rivers is still understudied and largely unquantified. In many freshwater ecosystems, Megaloptera larvae are a conspicuous benthic component, important in energy flow, recycling of materials, and food web dynamics.

Influenza A and B viruses possess a neuraminidase protein that shows sialidase activity. Influenza virus-specific neuraminidase inhibitors (NAIs) are commonly used for clinical treatment of influenza. However, some influenza A and B viruses that are resistant to NAIs have emerged in nature. NAI-resistant viruses have been monitored in public hygiene surveys and the mechanism underlying the resistance has been studied. Here, we describe a new assay for selective detection and isolation of an NAI-resistant virus in a speedy and easy manner by live fluorescence imaging of viral sialidase activity, which we previously developed, in order to achieve high-efficiency capture of an NAI-resistant virus. An NAI-resistant virus maintains sialidase activity even at a concentration of NAI that leads to complete deactivation of the virus. Infected cells and focuses (infected cell populations) of an oseltamivir-resistant virus were selectively visualized by live fluorescence sialidase imaging in the presence of oseltamivir, resulting in high-efficiency isolation of the resistant viruses. The use of a combination of other NAIs (zanamivir, peramivir, and laninamivir) in the imaging showed that the oseltamivir-resistant virus isolated in 2008 was sensitive to zanamivir and laninamivir but resistant to peramivir. Fluorescence imaging in the presence of zanamivir also succeeded in selective live-cell visualization of cells that expressed zanamivir-resistant NA. Fluorescence imaging of NAI-resistant sialidase activity will be a powerful method for study of the NAI resistance mechanism, for public monitoring of NAI-resistant viruses, and for development of a new NAI that shows an effect on various NAI-resistant mutations.

Fishflies (Corydalidae: Chauliodinae) are one of the main groups of the basal holometabolous insect order Megaloptera, with ca. 130 species distributed worldwide. A number of genera from the Southern Hemisphere show remarkably disjunctive distributions and are considered to be the austral remnants or \"living fossils\" of Gondwana. Hitherto, the evolutionary history of fishflies remains largely unexplored due to limited fossil record and incomplete knowledge of phylogenetic relationships. Here we describe two significant fossil species of fishflies, namely Eochauliodes striolatus gen. et sp. nov. and Jurochauliodes ponomarenkoi Wang & Zhang, 2010 (original designation for fossil larvae only), from the Middle Jurassic of Inner Mongolia, China. These fossils represent the earliest fishfly adults. Furthermore, we reconstruct the first phylogenetic hypothesis including all fossil and extant genera worldwide. Three main clades within Chauliodinae are recognized, i.e. the Dysmicohermes clade, the Protochauliodes clade, and the Archichauliodes clade. The phylogenetic and dispersal-vicariance (DIVA) analyses suggest Pangaean origin and global distribution of fishflies before the Middle Jurassic. The generic diversification of fishflies might have happened before the initial split of Pangaea, while some Gondwanan-originated clades were likely to be affected by the sequential breakup of Pangaea. The modern fauna of Asian fishflies were probably derived from their Gondwanan ancestor but not the direct descendents of the Mesozoic genera in Asia.