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Tardigrades can cope with adverse environmental conditions by turning into anhydrobiotes with a characteristic tun shape. Tun formation is an essential morphological adaptation for tardigrade entry into the anhydrobiotic state. The tun cell structure and ultrastructure have rarely been explored in tardigrades in general and never in Hypsibius exemplaris . We used transmission electron microscopy to compare cellular organization and ultrastructures between hydrated and anhydrobiotic H. exemplaris . Despite a globally similar cell organelle structure and a number of cells not significantly different between hydrated and desiccated tardigrades, reductions in the sizes of both cells and mitochondria were detected in dehydrated animals. Moreover, in anhydrobiotes, secretory active cells with a dense endoplasmic reticulum network were observed. Interestingly, these anhydrobiote-specific cells are in a close relationship with a specific extracellular structure surrounding each cell. It is possible that this rampart-like extracellular structure resulted from the accumulation of anhydrobiotic-specific material to protect the cells. Interestingly, after five hours of rehydration, the number of secretory cells decreased, and the specific extracellular structure began to disappear. Twenty-four hours after the beginning of rehydration, the cellular structure and ultrastructure were comparable to those observed in hydrated tardigrades.

Gryllus bimaculatus (Orthoptera: Gryllidae) is widely considered an excellent nutrient source for food and feed. Despite its economic importance, there is limited information on the impact of temperature on the bionomics of this cricket to guide its effective and sustainable mass production in its geographical range. The biological parameters of G . bimaculatus were investigated at eight different temperatures ranging from 20–40˚C. The Insect Life-Cycle Modelling (ILCYM) program was used to fit linear and non-linear functions to the data to describe the influence of temperature on life history parameters and its farmability under the current and projected climate for 2050. Our results revealed that G . bimaculatus was able to complete its lifecycle in the temperature range of 20°C to 37°C with a maximum finite rate of population increase (= 1.14) at 35°C. The developmental time of G . bimaculatus decreased with increasing temperature. The least developmental time and mortality were attained at 32°C. The highest wet length and mass of G . bimaculatus occurred at 32°C. The lowest temperature threshold for G . bimaculatus egg and nymph development was approximated using linear regression functions to be at 15.9°C and 16.2°C with a temperature constant of 108.7 and 555.6 degree days. The maximum fecundity (2301.98 eggs per female), net reproductive rate (988.42 daughters/ generation), and intrinsic rate of natural increase (0.134 days) were recorded at 32°C and the shortest doubling of 5.2 days was observed at 35°C. Based on our findings G . bimaculatus can be farmed in countries with temperatures ranging between 20 and 37°C around the globe. These findings will help the cricket farmers understand and project the cricket population dynamics around the world as influenced by temperature, and as such, will contribute to more efficient farming.

Although species delimitation is often controversial, emerging DNA-based and classical morphology-based methods are rarely compared using large-scale samplings, even less in the case of widely distributed species that have distant, allopatric populations. In the current study, we examined species boundaries within two wolf spider species of the genus Pardosa (Araneae, Lycosidae), P. riparia and P. palustris. Wolf spiders constitute an excellent model for testing the relevance of traditional vs. modern methods in species and population delimitation because several closely related species are distributed over cross-continental geographic ranges. Allopatric populations of the two Pardosa species were sampled across Europe to Far East Russia (latitudinal range > 150°) and several dozen individuals were studied using morphological characters (morphometry of three measures for both sexes, plus five in males only and two in females only), DNA barcoding (COI sequencing) and double-digest restriction site associated DNA sequencing (ddRADseq). The results obtained allow for changing the taxonomic status of two Far East Russian populations to subspecies and ddRADseq proved to be a powerful tool for taxonomic research despite scarce sampling and inherent subjectivity of species delimitation in allopatry. Overall, this study pleads for both multi-criteria and more population-based studies in taxonomy.

Leaves represent the main resource for herbivorous insects and their performances are mainly a function of leaf nutritional quality. Two feeding strategies are known to optimize the exploitation of leaf resources: leaf-miners that selectively feed on tissues of high nutritional quality and gall-inducers that induce the development of a new tissue showing an enhanced nutritional value. Some leaf-miners are known to also manipulate their nutritional environment, but do not affect plant development. Cases of callus proliferation in leaf-mines have been reported, however, the direct role of the insect in the formation of additional plant cells and the nutritional function of this tissue have never been established. Using an experimental approach, we show that leaf-mining larvae of micromoth, Borboryctis euryae (Lepidoptera: Gracillariidae), that grow on Eurya japonica (Pentaphylacaceae), actively induce callus proliferation within their leaf-mine at the fourth instar. We experimentally demonstrated that, at this developmental stage, the larva feeds exclusively on this newly formed tissue and feeding of the tissue is essential for completing larval stage. Phenological census revealed considerable expansion and variation of fourth instar duration caused by the continuous production of callus. We propose here the \"cornucopia\" hypothesis which states that the newly produced callus induced by the leaf-mining larvae provides virtually unending nourishment, which in turn allows flexible larval development time. This represents the first example of a leaf-miner manipulating plant development to its benefit, like a gall-inducer. We propose to name this life style \"mine-galler\".

In insects, the gall-inducing life-style has evolved independently many times. Several evolutionary pathways leading to this lifestyle have been proposed. While there is compelling evidence supporting surface-feeders and stem-borers as ancestral states of insect gall-inducers, an evolutionary pathway from leaf-miners remains hypothetical. Here we explored this question by comparing the developmental processes of two micromoths, a gall-inducer Caloptilia cecidophora (Lep., Gracillariidae), and its non-gall-inducing relative C. ryukyuensis. Like other Caloptilia, the first and second instars of C. cecidophora are leaf-miners and the gall is initiated inside the leaf mine by the third instar, thus suggesting leaf-mining as an ancestral, plesiomorphic state in this case. This is the first example of an insect species switching from leaf-mining to gall-inducing during larval development. The first two leaf-mining instars of C. cecidophora exhibit an absence of growth and a reduced time duration compared to C. ryukyuensis. The shortening of the duration of leaf-mining stages is apparently compensated in C. cecidophora by a larger egg size than C. ryukyuensis, and an additional larval instar during the gall phase.

The primary actors in the detection of olfactory information in insects are odorant receptors (ORs), transmembrane proteins expressed at the dendrites of olfactory sensory neurons (OSNs). In order to decode the insect olfactome, many studies focus on the deorphanization of ORs (i.e., identification of their ligand), using various approaches involving heterologous expression coupled to neurophysiological recordings. The “empty neuron system” of the fruit fly Drosophila melanogaster is an appreciable host for insect ORs, because it conserves the cellular environment of an OSN. Neural activity is usually recorded using labor-intensive electrophysiological approaches (single sensillum recordings, SSR). In this study, we establish a simple method for OR deorphanization using transcuticular calcium imaging (TCI) at the level of the fly antenna. As a proof of concept, we used two previously deorphanized ORs from the cotton leafworm Spodoptera littoralis , a specialist pheromone receptor and a generalist plant odor receptor. We demonstrate that by co-expressing the GCaMP6s/m calcium probes with the OR of interest, it is possible to measure robust odorant-induced responses under conventional microscopy conditions. The tuning breadth and sensitivity of ORs as revealed using TCI were similar to those measured using single sensillum recordings (SSR). We test and discuss the practical advantages of this method in terms of recording duration and the simultaneous testing of several insects.

Ontologies provide an important resource to integrate information. For developmental biology and comparative anatomy studies, ontologies of a species are used to formalize and annotate data that are related to anatomical structures, their lineage and timing of development. Here, we have constructed the first ontology for anatomy and asexual development (blastogenesis) of a bilaterian, the colonial tunicate Botryllus schlosseri. Tunicates, like Botryllus schlosseri, are non-vertebrates and the only chordate taxon species that reproduce both sexually and asexually. Their tadpole larval stage possesses structures characteristic of all chordates, i.e. a notochord, a dorsal neural tube, and gill slits. Larvae settle and metamorphose into individuals that are either solitary or colonial. The latter reproduce both sexually and asexually and these two reproductive modes lead to essentially the same adult body plan. The Botryllus schlosseri Ontology of Development and Anatomy (BODA) will facilitate the comparison between both types of development. BODA uses the rules defined by the Open Biomedical Ontologies Foundry. It is based on studies that investigate the anatomy, blastogenesis and regeneration of this organism. BODA features allow the users to easily search and identify anatomical structures in the colony, to define the developmental stage, and to follow the morphogenetic events of a tissue and/or organ of interest throughout asexual development. We invite the scientific community to use this resource as a reference for the anatomy and developmental ontology of B. schlosseri and encourage recommendations for updates and improvements.

Background Although whole-genome sequencing (WGS) is the preferred genotyping method for most genomic analyses, limitations are often experienced when studying genomes characterized by a high percentage of repetitive elements, high linkage, and recombination deserts. The Asian tiger mosquito ( Aedes albopictus ), for example, has a genome comprising up to 72% repetitive elements, and therefore we set out to develop a single-nucleotide polymorphism (SNP) chip to be more cost-effective. Aedes albopictus is an invasive species originating from Southeast Asia that has recently spread around the world and is a vector for many human diseases. Developing an accessible genotyping platform is essential in advancing biological control methods and understanding the population dynamics of this pest species, with significant implications for public health. Methods We designed a SNP chip for  Ae. albopictus (Aealbo chip) based on approximately 2.7 million SNPs identified using WGS data from 819 worldwide samples. We validated the chip using laboratory single-pair crosses, comparing technical replicates, and comparing genotypes of samples genotyped by WGS and the SNP chip. We then used the chip for a population genomic analysis of 237 samples from 28 sites in the native range to evaluate its usefulness in describing patterns of genomic variation and tracing the origins of invasions. Results Probes on the Aealbo chip targeted 175,396 SNPs in coding and non-coding regions across all three chromosomes, with a density of 102 SNPs per 1 Mb window, and at least one SNP in each of the 17,461 protein-coding genes. Overall, 70% of the probes captured the genetic variation. Segregation analysis found that 98% of the SNPs followed expectations of single-copy Mendelian genes. Comparisons with WGS indicated that sites with genotype disagreements were mostly heterozygotes at loci with WGS read depth < 20, while there was near complete agreement with WGS read depths > 20, indicating that the chip more accurately detects heterozygotes than low-coverage WGS. Sample sizes did not affect the accuracy of the SNP chip genotype calls. Ancestry analyses identified four to five genetic clusters in the native range with various levels of admixture. Conclusions The Aealbo chip is highly accurate, is concordant with genotypes from WGS with high sequence coverage, and may be more accurate than low-coverage WGS. Graphical Abstract

Host-parasite associations provide very useful models to study adaptive processes. We investigated the interaction between carnivorous marine gastropods, the Ovulidae or egg-cowries, and their cnidarian food targets. Ovulidae (Fleming, 1828), is a family of specialized carnivorous caenogastropods that feed by browsing on octocorals (Anthozoa: Octocorallia: Malacalcyonacea and Scleralcyonacea) or, to a much lesser degree, on antipatharians (Anthozoa: Hexacorallia: Antipatharia) and Stylasteridae (Hydrozoa: Hydroidolina: Anthoathecata). Very scanty information is available on the phylogenetic relationships and the degree of specificity of the relationship with the cnidarians of this corallivorous lineage, especially for deep-water taxa. To assess taxonomic identifications and investigate cnidarian/ovulid relationships in the context of their evolution, we generated an extensive molecular dataset comprising two mitochondrial (cox1 and 16S rDNA) and one nuclear gene (28S rDNA) from 524 specimens collected worldwide. The coral hosts of the ovulid species have been identified by integrating literature data with new records, employing morphological and/or molecular (the mitochondrial 16S rDNA and mtMSH, and the nuclear ITS2) markers. The obtained molecular phylogenetic framework for the Ovulidae revealed the existence of groups that do not completely correspond to the currently employed subfamilial arrangement. Concerning trophic ecology, while only pediculariines (Pedicularia and allied) are associated with hydrozoans (Stylasteridae), our results suggest that some ovulid lineages shifted independently

Calanoid copepods comprise around 90% of Arctic zooplankton biomass and are fundamental to the ecological and biogeochemical functioning of high-latitude pelagic ecosystems. They accumulate lipid reserves during the productive months and represent an energy-rich food source for higher trophic levels. Rapidly changing climate in the Arctic may alter the quantity and composition of the food environment for one of the key copepod species, Calanus finmarchicus , with as yet unquantified effects on its production. Here we present rates of feeding and egg production in female C. finmarchicus exposed to the range of feeding conditions encountered across the Fram Strait in May/June 2018. Carbon (C) budgets were constructed and used to examine the relationship between feeding and growth (= egg production) in these animals. C-specific ingestion rates (mean ± standard deviation) were highly variable, ranging from 0.015 ± 0.004 to 0.645 ± 0.017 day -1 (mean = 0.295 ± 0.223 day -1 ), and were positively correlated with food availability. C-specific egg production rates ranged from 0.00 to 0.049 day -1 (mean = 0.012 ± 0.011) and were not correlated with either food availability or ingestion rate. Calculated gross growth efficiencies (GGE: growth/ingestion) were low, 0.12 ± 0.13 (range = 0.01 to 0.39). The assembled C budgets indicate that the average fraction of ingested food that was surplus to the requirements for egg production, respiration and losses to faecal pellets was 0.17 ± 0.42. We suggest that this excess occurred, at least in part, because many of the incubated females were still undergoing the energetically (C-) expensive process of gonad maturation at the time of sampling, an assertion that is supported by the relatively high C:N (nitrogen) ratios of the incubated females, the typically low egg production rates, and gonad maturation status. Ontogenetic development may thus explain the large variability seen in the relationship between egg production and ingestion. The apparently excessive ingestion rates may additionally indicate that recently moulted females must acquire additional N via ingestion to complete the maturation process and begin spawning. Our results highlight the need for improved fundamental understanding of the physiology of high-latitude copepods and its response to environmental change.