The genome sequence of a cased caddisfly, Mystacides longicornis (Linnaeus, 1758).

We present a genome assembly from an individual male Mystacides longicornis (cased caddisfly; Arthropoda; Insecta; Trichoptera; Leptoceridae). The genome sequence is 665.1 megabases in span. Most of the assembly is scaffolded into 20 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 15.75 kilobases in length.

Phylogenomics recovers multiple origins of portable case making in caddisflies (Insecta: Trichoptera), nature's underwater architects.

Caddisflies (Trichoptera) are among the most diverse groups of freshwater animals with more than 16 000 described species. They play a fundamental role in freshwater ecology and environmental engineering in streams, rivers and lakes. Because of this, they are frequently used as indicator organisms in biomonitoring programmes. Despite their importance, key questions concerning the evolutionary history of caddisflies, such as the timing and origin of larval case making, remain unanswered owing to the lack of a well-resolved phylogeny. Here, we estimated a phylogenetic tree using a combination of transcriptomes and targeted enrichment data for 207 species, representing 48 of 52 extant families and 174 genera. We calibrated and dated the tree with 33 carefully selected fossils. The first caddisflies originated approximately 295 million years ago in the Permian, and major suborders began to diversify in the Triassic. Furthermore, we show that portable case making evolved in three separate lineages, and shifts in diversification occurred in concert with key evolutionary innovations beyond case making.

Bigger is not necessarily better: empirical tests show that dispersal proxies misrepresent actual dispersal ability.

Tests for the role of species' relative dispersal abilities in ecological and biogeographical models rely heavily on dispersal proxies, which are seldom substantiated by empirical measures of actual dispersal. This is exemplified by tests of dispersal-range size relationships and by metacommunity research that often features invertebrates, particularly freshwater insects. Using rare and unique empirical data on dispersal abilities of caddisflies, we tested whether actual dispersal abilities were associated with commonly used dispersal proxies (metrics of wing size and shape; expert opinion). Across 59 species in 12 families, wing morphology was not associated with actual dispersal. Within some families, individual wing metrics captured some dispersal differences among species, although useful metrics varied among families and predictive power was typically low. Dispersal abilities assigned by experts were either no better than random or actually poorer than random. Our results cast considerable doubt on research underpinned by dispersal proxies and scrutiny of previous research results may be warranted. Greater progress may lie in employing innovative survey and experimental design to measure actual dispersal in the field.

The silk gland proteome of Stenopsyche angustata provides insights into the underwater silk secretion.

Caddisworms (Trichoptera) spin adhesive silks to construct a variety of underwater composite structures. Many studies have focused on the fibroin heavy chain of caddisworm silk and found that it contains heavy phosphorylation to maintain a stable secondary structure. Besides fibroins, recent studies have also identified some new silk proteins within caddisworm silk. To better understand the silk composition and its secretion process, this study reports the silk gland proteome of a retreat-building caddisworm, Stenopsyche angustata Martynov (Trichoptera, Stenopsychidae). Using liquid chromatography tandem mass spectrometry (LC-MS/MS), 2389 proteins were identified in the silk gland of S. angustata, among which 192 were predicted as secreted silk proteins. Twenty-nine proteins were found to be enriched in the front silk gland, whereas 109 proteins were enriched in the caudal silk gland. The fibroin heavy chain and nine uncharacterized silk proteins were identified as phosphorylated proteins. By analysing the sequence of the fibroin heavy chain, we found that it contains 13 Gly/Thr/Pro-rich regions, 12 Val/Ser/Arg-rich regions and a Gly/Arg/Thr-rich region. Three uncharacterized proteins were identified as sericin-like proteins due to their larger molecular weights, signal peptides and repetitive motifs rich in serine. This study provides valuable information for further clarifying the secretion and adhesion of underwater caddisworm silk.

New species of the genus Trichosetodes Ulmer, 1915 (Trichoptera, Leptoceridae) from Ratanakiri province, Cambodia, based on morphological and molecular data.

Three new species of Trichosetodes, namely T.carmelaesp. nov., T.katiengensissp. nov. and T.ratanakiriensissp. nov. are described and illustrated by male specimens. The male genitalia of T.carmelaesp. nov. can be distinguished from the other 16 species of the genus found in Southeast Asia by the shape of the phallicata. The phallicata of T.carmelaesp. nov. bears a tuft of long hairs in the middle of the dorsal edge. Trichosetodeskatiengensissp. nov. can be distinguished from the other species in Southeast Asia by the shape of the phallicata which is divided into dorsal and ventral branches in lateral view, and T.ratanakiriensissp. nov. by the characters of the left inferior appendage and the shape of segment IX. The posterior end of the left inferior appendage of T.ratanakiriensissp. nov. is not forked and the ventral and lateral views of the posteroventral lobes of segment IX are rounded. Illustrations of male genitalia of Trichosetodeskampongspeuensis Malicky & Kong, 2020 are provided for comparison. The molecular diversity of new Trichosetodes species was analyzed using the mitochondrial large subunit ribosomal rRNA gene region (16S rRNA). In terms of their genetic divergence, T.ratanakiriensissp. nov. and T.kampongspeuensis exhibited remarkable proximity, with only a 1.4% distance. On the contrary, T.carmelaesp. nov. displayed genetic disparity exceeding 6.3% when compared to both T.ratanakiriensissp. nov. and T.kampongspeuensis.

The genome sequence of the cinnamon sedge caddisfly, Limnephilus marmoratus (Curtis, 1834).

We present a genome assembly from an individual Limnephilus marmoratus (a caddisfly; Arthropoda; Insecta; Trichoptera; Limnephilidae). The genome sequence is 1,630 megabases in span. Most of the assembly (99.93%) is scaffolded into 30 chromosomal pseudomolecules, including the assembled Z sex chromosome. The mitochondrial genome has also been assembled and is 15.4 kilobases in length.

The genome sequence of a caddisfly, Limnephilus lunatus (Curtis, 1834).

We present a genome assembly from an individual Limnephilus lunatus (a caddisfly; Arthropoda; Insecta; Trichoptera; Limnephilidae). The genome sequence is 1,270 megabases in span. Most of the assembly is scaffolded into 13 chromosomal pseudomolecules, including the assembled Z chromosome. The mitochondrial genome has also been assembled and is 15.4 kilobases long.

Highly accurate long reads are crucial for realizing the potential of biodiversity genomics.

BACKGROUND: Generating the most contiguous, accurate genome assemblies given available sequencing technologies is a long-standing challenge in genome science. With the rise of long-read sequencing, assembly challenges have shifted from merely increasing contiguity to correctly assembling complex, repetitive regions of interest, ideally in a phased manner. At present, researchers largely choose between two types of long read data: longer, but less accurate sequences, or highly accurate, but shorter reads (i.e., >Q20 or 99% accurate). To better understand how these types of long-read data as well as scale of data (i.e., mean length and sequencing depth) influence genome assembly outcomes, we compared genome assemblies for a caddisfly, Hesperophylax magnus, generated with longer, but less accurate, Oxford Nanopore (ONT) R9.4.1 and highly accurate PacBio HiFi (HiFi) data. Next, we expanded this comparison to consider the influence of highly accurate long-read sequence data on genome assemblies across 6750 plant and animal genomes. For this broader comparison, we used HiFi data as a surrogate for highly accurate long-reads broadly as we could identify when they were used from GenBank metadata. RESULTS: HiFi reads outperformed ONT reads in all assembly metrics tested for the caddisfly data set and allowed for accurate assembly of the repetitive ~ 20 Kb H-fibroin gene. Across plants and animals, genome assemblies that incorporated HiFi reads were also more contiguous. For plants, the average HiFi assembly was 501% more contiguous (mean contig N50 = 20.5 Mb) than those generated with any other long-read data (mean contig N50 = 4.1 Mb). For animals, HiFi assemblies were 226% more contiguous (mean contig N50 = 20.9 Mb) versus other long-read assemblies (mean contig N50 = 9.3 Mb). In plants, we also found limited evidence that HiFi may offer a unique solution for overcoming genomic complexity that scales with assembly size. CONCLUSIONS: Highly accurate long-reads generated with HiFi or analogous technologies represent a key tool for maximizing genome assembly quality for a wide swath of plants and animals. This finding is particularly important when resources only allow for one type of sequencing data to be generated. Ultimately, to realize the promise of biodiversity genomics, we call for greater uptake of highly accurate long-reads in future studies.

Four New Species of the Genus Eoneureclipsis (Trichoptera: Psychomyiidae) from China Inferred from Morphology and DNA Barcodes.

Four new species of the genus Eoneureclipsis Kimmins, 1955 from China are described, illustrated, and diagnosed based on male genitalia: Eoneureclipsis jianfenglingensis sp. nov. from Hainan, E. foraminulatus sp. nov. from Guangxi, E. spinosus sp. nov. from Guangxi and Guangdong, and E. gei sp. nov. from Fujian. A dichotomous key to Chinese adult males of Eoneureclipsis is provided. A distribution map for all Eoneureclipsis species is also presented. The DNA barcodes (partial mtCOI sequences) of E. jianfenglingensis sp. nov., E. gei sp. nov., and E. hainanensis Mey, 2013 have been generated and compared with all existing sequences of Eoneureclipsis species.

The genome sequence of a caddisfly, Limnephilus auricula (Curtis, 1834).

We present a genome assembly from an individual female Limnephilus auricula (a caddisfly; Arthropoda; Insecta; Trichoptera; Limnephilidae). The genome sequence is 971.3 megabases in span. Most of the assembly is scaffolded into 30 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled and is 18.29 kilobases in length.

A new species of Nectopsyche Müller, 1879 (Trichoptera: Leptoceridae) and notes on the adults of Nectopsyche splendida ()

ABSTRACT The male and female of a new species in the long-horned caddisfly genus Nectopsyche Müller, 1879 are described from southeastern Brazil. The new species has male genitalia similar to species of the candida-group, but has several endothecal spines in the phallic apparatus similar to those of gemma-group species. The color pattern of the scales and hairs on the head, thorax and forewings is similar to that of Nectopsyche splendida (Navás, 1917), as both species have bands of metallic silver-iridescent scales interleaved with black areas on the forewings. Additionally, we present illustrations of the male and female genitalia of N. splendida and expand the knowledge of its distribution.

High Performance Marine and Terrestrial Bioadhesives and the Biomedical Applications They Have Inspired.

This study has reviewed the naturally occurring bioadhesives produced in marine and freshwater aqueous environments and in the mucinous exudates of some terrestrial animals which have remarkable properties providing adhesion under difficult environmental conditions. These bioadhesives have inspired the development of medical bioadhesives with impressive properties that provide an effective alternative to suturing surgical wounds improving closure and healing of wounds in technically demanding tissues such as the heart, lung and soft tissues like the brain and intestinal mucosa. The Gecko has developed a dry-adhesive system of exceptional performance and has inspired the development of new generation re-usable tapes applicable to many medical procedures. The silk of spider webs has been equally inspiring to structural engineers and materials scientists and has revealed innovative properties which have led to new generation technologies in photonics, phononics and micro-electronics in the development of wearable biosensors. Man made products designed to emulate the performance of these natural bioadhesive molecules are improving wound closure and healing of problematic lesions such as diabetic foot ulcers which are notoriously painful and have also found application in many other areas in biomedicine. Armed with information on the mechanistic properties of these impressive biomolecules major advances are expected in biomedicine, micro-electronics, photonics, materials science, artificial intelligence and robotics technology.

New Mitogenome Features of Philopotamidae (Insecta: Trichoptera) with Two New Species of Gunungiella.

A total of 14 individuals of Philopotamidae, from China, were examined. Six species in four genera, including two new species of the genus Gunungiella, were recognized. Their COI barcode sequences were extracted, mitogenomes were sequenced, assembled and analyzed. All of these sequences were used to further reveal the phylogenetic relationships of the family Philopotamidae. In addition, two new species: Gunungiella wangi n. sp., Gunungiella flabellata n. sp. were described and illustrated.

Caddisfly Larvae are a Driver of Plastic Litter Breakdown and Microplastic Formation in Freshwater Environments.

Plastic litter is now pervasive in the aquatic environment. Several marine and terrestrial organisms can fragment plastic with their feeding appendages, facilitating its breakdown and generating microplastics. However, similar studies with freshwater organisms are extremely limited. We explored the interactions between the caddisfly larvae Agrypnia sp. and polylactic acid (PLA) film. The use of plastic by larvae to build their protective cases was investigated, along with their ability to fragment the plastic film as they do with leaf litter. Caddisfly consistently incorporated PLA into their cases alongside leaf material. They also used their feeding appendages to rapidly fragment PLA-forming hundreds of submillimeter-sized microplastics. Although larvae showed a preference for leaf material when constructing cases, plastic use and fragmentation still occurred when leaf material was replete, indicating that this behavior is likely to occur in natural environments that are polluted with plastics. This is thought to be the first documented evidence of active plastic modification by a freshwater invertebrate and therefore reveals a previously unidentified mechanism of plastic fragmentation and microplastic formation in freshwater. Further work is now needed to determine the extent of this behavior across freshwater taxa and the potential implications for the wider ecosystem. Environ Toxicol Chem 2022;41:3058-3069. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Mild temperatures differentiate while extreme temperatures unify gene expression profiles among populations of Dicosmoecus gilvipes in California.

Accurately predicting the effects of future warming on aquatic ectotherms requires an understanding how thermal history, including average temperature and variation, affects populations of the same species. However, many laboratory studies simplify the thermal environment to focus on specific organismal responses and sacrifice environmental realism. Here, we paired laboratory-based transcriptomic RNA-seq analysis to identify thermally responsive genes with NanoString analysis of a subset of those genes to characterize natural field-based variation in thermal physiology among populations. We tested gene expression responses of three populations of field-acclimatized larval caddisflies (Dicosmoecus gilvipes) from streams in different eco-regions (mountain, valley, and coast) following exposure to current and future summertime temperatures. We hypothesized that distinct thermal histories across eco-regions could differentiate populations at baseline "control" levels of gene expression, as well as gene expression changes in response to daily warming and heat shock. Population-specific patterns of gene expression were apparent under the control and daily warming conditions suggesting that local acclimatization or local adaptation may differentiate populations, while responses to extreme temperatures were similar across populations, indicating that response to thermal stress is canalized. Underlying gene co-expression patterns in the daily warming and heat shock treatments were different, demonstrating the distinct physiological mechanisms involved with thermal acclimatization and response to thermal stress. These results highlight the importance and limitations of studies of the thermal biology of wild-caught organisms in their natural environment, and provide an important resource for researchers of caddisflies and aquatic insects in general.

Using the multi-omics approach to reveal the silk composition in Plectrocnemia conspersa.

Similar to Lepidoptera, the larvae of Trichoptera are also capable of producing silk. Plectrocnemia conspersa, a predatory species belonging to the suborder Annulipalpia, builds massive silken retreats with preycapturing nets. In this study, we describe the silk glands of P. conspersa and use the multi-omics methods to obtain a complete picture of the fiber composition. A combination of silk gland-specific transcriptome and proteomic analyses of the spun-out fibers yielded 27 significant candidates whose full-length sequences and gene structures were retrieved from the publicly available genome database. About one-third of the candidates were completely novel proteins for which there are no described homologs, including a group of five pseudofibroins, proteins with a composition similar to fibroin heavy chain. The rest were homologs of lepidopteran silk proteins, although some had a larger number of paralogs. On the other hand, P. conspersa fibers lacked some proteins that are regular components in moth silk. In summary, the multi-omics approach provides an opportunity to compare the overall composition of silk with other insect species. A sufficient number of such studies will make it possible to distinguish between the basic components of all silks and the proteins that represent the adaptation of the fibers for specific purposes or environments.

Population regulation throughout a complex life cycle.

Research Highlight: McIntosh, A. R., Greig, H. S., & Howard, S. (2022). Regulation of open populations of a stream insect through larval density dependence. Journal of Animal Ecology. https://doi.org/10.1111/1365-2656.13696. Despite decades of research on population regulation through density dependence, it remains challenging to identify and understand the relative importance of mechanisms governing open populations of organisms with complex life cycles. McIntosh et al. (2022) manipulated density of aquatic invertebrates in the field, and then followed populations for 2 years to track the effects on abundance through multiple life-history stages. The authors found that their density manipulation, performed on larvae that were about to pupate, had minimal effects on the number of emergent adults collected several months later. The manipulation had a similarly negligible influence on the number of egg masses laid at study locations. The authors attribute this to stochasticity around dispersal of flying adults through the terrestrial environment. However, later in the study, the authors found evidence of density-dependent population regulation among larval stages, seemingly controlled by resource availability. These results suggest that population dynamics depend on multiple mechanisms operating at different points in organisms' life history, which could either help or hinder population persistence with disturbance or environmental change.

Associating life stages and sexes of Nearctic Polycentropus Curtis, 1835 (Trichoptera: Polycentropodidae) using mitochondrial DNA barcoding.

Alpha taxonomy of caddisflies (order Trichoptera) is based primarily on male genital morphology. As such, associations of adult females and other life stages typically require conclusive association with the species' identifiable male. The aim of this study was to use molecular methods to associate females and larvae of Polycentropus species represented in the Nearctic. Analysis of mtCOI sequences using distance- and tree-based methods resulted in the association of larvae for 14 species of Polycentropus (P. alabamensis Hamilton, Harris & Lago, 1990, P. blicklei Ross & Yamamoto 1965, P. carlsoni Morse 1971, P. carolinensis Banks 1905, P. colei Ross 1941, P. confusus Hagen 1861, P. denningi Smith 1962, P. elarus Ross 1944, P. gertschi Denning 1950, Polycentropus halidus Milne 1936, P. maculatus Banks 1908, P. pentus Ross 1941, P. rickeri Yamamoto 1966, and P. variegatus Banks 1900) and females for 2 species (P. carolinensis and P. chelatus Ross & Yamamoto 1965). Searches for, and descriptions of, diagnostic morphological characters for these previously unidentifiable life forms are now possible. The identity of the larva of P. centralis Banks, 1914 is confirmed and some interesting phylogenetic relationships and a possible cryptic species and potential synonyms are implied in the results. Targets for future immature- and female-male associations are discussed along with a preliminary assessment of morphological differences among larvae.

The return to land: association between hairworm infection and aquatic insect development.

Host manipulation by parasites can shape host behaviour, community structure, and the flow of energy through food webs. A well-known example of host manipulation comes from hairworms (phylum Nematomorpha), which somehow cause their terrestrial insect definitive hosts to enter water, a phenomenon that has received lots of attention in recent years. However, little focus has been directed towards the interactions between hairworms and their aquatic insect hosts and the return of dormant hairworms from water to land. Here, we ask whether hairworm cyst infections impact, either directly or indirectly, the life history of their aquatic transport hosts. By observing the development of last-instar Olinga jeanae (Trichoptera: Conoesucidae) caddisfly larvae naturally infected with Gordius-type hairworm cysts under controlled conditions, we found that higher numbers of cysts per infected caddisfly correlated with a decrease in time to pupation. These new observations suggest that, apart from the striking host manipulation that brings the parasite from land to water, the presence of dormant hairworms is associated with changes in the development of their aquatic hosts, either through direct or indirect mechanisms, which may accelerate their transition from water to land.

The First Report of the Genera Abaria and Drepanocentron (Trichoptera: Xiphocentronidae) from China, with Descriptions of Two New Species.

The genera Abaria Mosely 1948 and Drepanocentron Schmid 1982 are recorded in China for the first time. In this study, two new species, Abaria herringbona sp. nov., from Guang-xi, and Drepanocentron fuxiensis sp. nov., from An-hui, are described and illustrated. Male genitalia of these two new species is distinguishable from those of other Abaria and Drepanocentron species. In addition, Melanotrichia attia Malicky & Chantaramongkol 1992 is a new record for the Chinese caddisfly fauna.