Insects are the most speciose group of animals, but the phylogenetic relationships of many major lineages remain unresolved. We inferred the phylogeny of insects from 1478 protein-coding genes. Phylogenomic analyses of nucleotide and amino acid sequences, with site-specific nucleotide or domain-specific amino acid substitution models, produced statistically robust and congruent results resolving previously controversial phylogenetic relations hips. We dated the origin of insects to the Early Ordovician [~479 million years ago (Ma)], of insect flight to the Early Devonian (~406 Ma), of major extant lineages to the Mississippian (~345 Ma), and the major diversification of holometabolous insects to the Early Cretaceous. Our phylogenomic study provides a comprehensive reliable scaffold for future comparative analyses of evolutionary innovations among insects.

Wastewater-based monitoring of the spread of the new SARS-CoV-2 virus, also referred to as wastewater-based epidemiology (WBE), has been suggested as a tool to support epidemiology. An extensive sampling campaign, including nine municipal wastewater treatment plants, has been conducted in different cities of the Federal State of North Rhine-Westphalia (Germany) on the same day in April 2020, close to the first peak of the corona crisis. Samples were processed and analysed for a set of SARS-CoV-2-specific genes, as well as pan-genotypic gene sequences also covering other coronavirus types, using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Additionally, a comprehensive set of chemical reference parameters and bioindicators was analysed to characterize the wastewater quality and composition. Results of the RT-qPCR based gene analysis indicate the presence of SARS-CoV-2 genetic traces in different raw wastewaters. Furthermore, selected samples have been sequenced using Sanger technology to confirm the specificity of the RT-qPCR and the origin of the coronavirus. A comparison of the particle-bound and the dissolved portion of SARS-CoV-2 virus genes shows that quantifications must not neglect the solid-phase reservoir. The infectivity of the raw wastewater has also been assessed by viral outgrowth assay with a potential SARS-CoV-2 host cell line in vitro, which were not infected when exposed to the samples. This first evidence suggests that wastewater might be no major route for transmission to humans. Our findings draw attention to the need for further methodological and molecular assay validation for enveloped viruses in wastewater.

Background The latest advancements in DNA sequencing technologies have facilitated the resolution of the phylogeny of insects, yet parts of the tree of Holometabola remain unresolved. The phylogeny of Neuropterida has been extensively studied, but no strong consensus exists concerning the phylogenetic relationships within the order Neuroptera. Here, we assembled a novel transcriptomic dataset to address previously unresolved issues in the phylogeny of Neuropterida and to infer divergence times within the group. We tested the robustness of our phylogenetic estimates by comparing summary coalescent and concatenation-based phylogenetic approaches and by employing different quartet-based measures of phylogenomic incongruence, combined with data permutations. Results Our results suggest that the order Raphidioptera is sister to Neuroptera + Megaloptera. Coniopterygidae is inferred as sister to all remaining neuropteran families suggesting that larval cryptonephry could be a ground plan feature of Neuroptera. A clade that includes Nevrorthidae, Osmylidae, and Sisyridae (i.e. Osmyloidea) is inferred as sister to all other Neuroptera except Coniopterygidae, and Dilaridae is placed as sister to all remaining neuropteran families. Ithonidae is inferred as the sister group of monophyletic Myrmeleontiformia. The phylogenetic affinities of Chrysopidae and Hemerobiidae were dependent on the data type analyzed, and quartet-based analyses showed only weak support for the placement of Hemerobiidae as sister to Ithonidae + Myrmeleontiformia. Our molecular dating analyses suggest that most families of Neuropterida started to diversify in the Jurassic and our ancestral character state reconstructions suggest a primarily terrestrial environment of the larvae of Neuropterida and Neuroptera. Conclusion Our extensive phylogenomic analyses consolidate several key aspects in the backbone phylogeny of Neuropterida, such as the basal placement of Coniopterygidae within Neuroptera and the monophyly of Osmyloidea. Furthermore, they provide new insights into the timing of diversification of Neuropterida. Despite the vast amount of analyzed molecular data, we found that certain nodes in the tree of Neuroptera are not robustly resolved. Therefore, we emphasize the importance of integrating the results of morphological analyses with those of sequence-based phylogenomics. We also suggest that comparative analyses of genomic meta-characters should be incorporated into future phylogenomic studies of Neuropterida.

Family interrelationships among Anisoptera (dragonflies) are unresolved. Molecular markers applied thus far have not been particularly useful for resolving relationships at the family level. Previous morphological studies have depended heavily on characters of wing venation and articulation which are believed to display considerable degrees of homoplasy due to adaptations to different flight modes. Here, we present a comprehensive anatomical dataset of the head morphology of Anisoptera focusing on muscle organization and endoskeletal features covering nearly all families. The characters are illustrated in detail and incorporated into an updated morphological character matrix covering all parts of the dragonfly body. Phylogenetic analysis recovers all families as monophyletic clades except Corduliidae, Gomphidae as sister group to all remaining Anisoptera, and Austropetaliidae as sister group to Aeshnidae (=Aeshnoidea). The position of Petaluridae and Aeshnoidea to each other could not be resolved. Libelluloidea is monophyletic with Neopetalia and Cordulegastridae as first splits. Chlorogomphidae is sister to monophyletic [Synthemistidae + (‘Corduliidae’ + Libellulidae)]. In addition, we applied a recently published formal approach to detect concerted convergence in morphological data matrices and uncover possible homoplasies. Analyses show that especially head and thorax characters may harbour homoplasies. After exclusion of possible homoplastic characters, Gomphidae is corroborated as sister group to all remaining Anisoptera.

Since its discovery and description, the oriental moth Pseudobiston pinratanai Inoue, 1994, could not be placed in any of the recognized families of Macroheterocera (Lepidoptera). Here, we used molecular and morphological data to infer the phylogenetic position of the species. These analyses indicate that Pseudobiston pinratanai is closely related to Epicopeiidae. Our analyses of morphological characters further show that Pseudobiston pinratanai+Epicopeiidae share synapomorphies with Sematuridae, Uraniidae and Geometridae. These observations lead us to redefine the superfamily Geometroidea and discuss its internal phylogenetic relationships. The isolated position of Pseudobiston pinratanai within Geometroidea supports a family status (Pseudobistonidae fam. n.) that we formally establish in this work.