Search

The Antarctic midge, Belgica antarctica, is a unique insect endemic to Antarctica. It has a 2-year life cycle, with larvae overwintering in two different instars and adults emerging the following summer. This seasonality is crucial for adaptation to Antarctica's harsh climates and ephemeral growing seasons; however, the underlying mechanisms remain unclear. We found that, under summer-like conditions, larvae could develop from egg to the fourth-instar larval stage without interruption, but they never pupated. Spontaneous developmental arrest at this stage suggests that they overwinter in obligate diapause, a genetically determined period of dormancy. The winter cold can terminate this diapause, and long-term cold exposure is more effective. Although this species can utilise two alternative cold tolerance strategies with diapause for overwintering, freezing was more successful than cryoprotective dehydration in allowing survival and developmental resumption in our experimental conditions. In contrast, the first three larval instars continued their development under the same conditions as the fourth-instar larvae. Although we do not exclude the possibility of facultative diapause, they likely overwinter in a quiescent state, an immediate developmental arrest in response to adversity, to maximise exploitation of the short Antarctic summer. Diapause and quiescence ensure developmental and reproductive success in this extremophile insect., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Insect gut symbiotic microbiota play a crucial role in the nutritional, physiological, and behavioral aspects of their hosts, providing valuable insights for investigating the co-evolution of insects and plants. Sophora alopecuroides L. serves as an important windbreak plant, while Etiella zinckenella is a major pest that infests its seeds. However, the structure of the gut microbiota community in E. zinckenella remains poorly understood. In this study, we analyzed the gut microbiota of E. zinckenella across different developmental stages-larvae (1st-5th instars), pupae, and adults-infesting S. alopecuroides using 16 S rRNA high-throughput sequencing. The results revealed that the dominant phyla throughout the development of E. zinckenella were Proteobacteria and Bacteroidota, although the dominant genera varied significantly across stages. Diversity analysis of gut microbiota at different developmental stages indicated that microbial diversity was significantly higher in the larval stage compared to the pupal and adult stages. Functional predictions further highlighted the richness of metabolic pathways within the gut microbiota of E. zinckenella. Notably, carbohydrate metabolism functions were significantly more abundant during the larval stage, while lipid metabolism functions were substantially lower. Our findings demonstrate dynamic changes in the composition and diversity of the gut microbiota across the developmental stages of E. zinckenella, underscoring the critical roles of these bacteria during specific stages of the insect's life cycle. This study lays the groundwork for future strategies aimed at controlling E. zinckenella through modulation of its gut microbiota, offering significant theoretical implications., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Tomato (Lycopersicon esculentum Miller), is a globally important agricultural product, yet it is under significant threat from pests such as the tomato leaf miner, Phthorimaea absoluta (Meyrick) (Lepidoptera: Gelechiidae). This study investigates the combined effects of gibberellin and vermiwash treatment on the life history and antioxidant system of P. absoluta. Given the pest's resistance to many chemical pesticides, alternative control methods are crucial. Gibberellins are plant growth hormones known for their role in plant development and stress responses, while vermiwash is a bio-fertilizer rich in nutrients and microbial agents. We applied gibberellin and vermiwash (GV treatment) to tomato plants and assessed the impact on P. absoluta developmental stages, reproduction, and enzymatic activities. Our results show significant differences in larval development times (32.06 ± 0.39) and survival rates (0.53 ± 0.09) between treated and control groups (27.38 ± 0.35 and 0.80 ± 0.07, respectively). The GV treatment prolonged the total lifespan of P. absoluta (44.31 ± 0.51) but reduced its intrinsic rate of increase (r) (0.086 ± 0.009) and finite rate of increase (λ) (1.090 ± 0.009). Enzymatic assays revealed altered antioxidant and detoxifying enzyme activities in treated larvae. This study suggests that gibberellin and vermiwash treatments could be incorporated into pest management strategies for sustainable tomato production., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

The red king crab, Paralithodes camtschaticus, and the Japanese mitten crab, Eriocheir japonica, are the major commercially valuable species. In addition to their high nutritional value, these crabs are used as objects of ecological research. To extend our knowledge of crustacean biochemistry and provide a more comprehensive model of lipidomic patterns during embryonic and larval development of these crab species, we studied the dynamics of molecular species profiles of reserve lipids such as triacylglycerols (TG) and membrane lipids such as glycerophospholipids (PL). A complete disappearance of TG was observed in zoea IV larvae of E. japonica and zoea III larvae of P. camtschaticus. The appearance of TG at older stages of larval development was accompanied by considerable changes in TG composition. The dynamics of PL with major polyunsaturated fatty acids (PUFA) (20:5n-3, 22:6n-3, and 20:4n-3) during the larval development was species-specific. The obtained results indicate different demands for PUFA in P. camtschaticus and E. japonica, which can be taken into account when selecting optimum diets. The lipidomic approach allows identifying new patterns of lipid changes during crab embryonic development, which may be useful for improvement of aquaculture techniques., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

The fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae) is a major phytophagous pest that invaded China in late 2018, posing a serious threat to local agricultural production. Therefore, we investigated the effects of maize, soybean, and sweet potato on the growth, development, and reproduction of S. frugiperda under laboratory conditions. The developmental period of the egg-larval stage was significantly longer when S. frugiperda fed on sweet potato (28.94 days) compared to maize (16.19 days) and soybean (17.82 days). Sweet potato feeding significantly prolonged the pupal period, but this effect was not observed in the adult stage. Spodoptera frugiperda larvae fed on sweet potato had the lowest pupal weight (116.18 mg) and pupation rate (68.19%). The mean fecundity of females significantly differed among the plants, with egg production being highest for insects fed on maize (996.17 eggs) and lowest for those fed on sweet potato (319.28 eggs). Spodoptera frugiperda fed on sweet potato exhibited the smallest net reproductive rate (47.892), lowest intrinsic rate of increase (0.083 day -1 ), lowest finite rate of increase (1.086 day -1 ), and longest mean generation time (46.806 days). Overall, S. frugiperda can survive and complete its entire life cycle on all three host plants., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Tables., (© 2024. The Author(s).)

The sterile insect technique (SIT) is a sustainable genetic control method to effectively suppress mosquito vector populations below sanitary and nuisance thresholds. The SIT requires a cost-effective production and release of radio sterilized insects of high and consistent quality and quantity. In this study, we report a new procedure to hatch, aliquot and introduce Aedes albopictus and Aedes aegypti larvae into larval mass rearing units using dissolvable vegetable capsules containing brushed eggs and larval diet. The results demonstrated that the egg brushing procedures, the opening of the hatching container and the presence of diet in the hatching solution increased Aedes eggs hatching success. The diet added during the hatching process decreased the dissolved oxygen (DO) concentration, which constituted the main stimulus for egg hatching success in both species. However, a DO reduction below 3 mg/l can effectively stimulate hatching in both species regardless of the deoxygenation method adopted. The capsule method produced similar hatching success without affecting larval survival and pupation rates in comparison with standard hatching procedures. The use of capsules eliminates intermediate laborious procedures for hatching, counting and transferring larvae into mass rearing trays with an expected improvement in production times and costs for Aedes SIT operational programs., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Anurans are characterized by a biphasic life cycle, with an aquatic larval (tadpole) stage followed by an adult (frog) stage, both connected through the metamorphic period in which drastic morphological and physiological changes occur 1 . Extant tadpoles exhibit great morphological diversity and ecological relevance 2 , but their absence in the pre-Cretaceous fossil record (older than 145 million years) makes their origins and early evolution enigmatic. This contrasts with the postmetamorphic anuran fossil record that dates back to the Early Jurassic and with closely related species in the Late Triassic (around 217-213 million years ago (Ma)) 3 . Here we report a late-stage tadpole of the stem-anuran Notobatrachus degiustoi from the Middle Jurassic of Patagonia (around 168-161 Ma). This finding has dual importance because it represents the oldest-known tadpole and, to our knowledge, the first stem-anuran larva. Its exquisite preservation, including soft tissues, shows features associated with the filter-feeding mechanism characteristic of extant tadpoles 4,5 . Notably, both N. degiustoi tadpole and adult reached a large size, demonstrating that tadpole gigantism occurred among stem-anurans. This new discovery reveals that a biphasic life cycle, with filter-feeding tadpoles inhabiting aquatic ephemeral environments, was already present in the early evolutionary history of stem-anurans and has remained stable for at least 161 million years., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

MicroRNAs (miRNAs) regulate a wide variety of biological processes by silencing their target genes. Argonaute (AGO) proteins load miRNAs to form an RNA-induced silencing complex (RISC), which mediates translational repression and/or mRNA decay of the targets. A scaffold protein called GW182 directly binds AGO and the CCR4-NOT deadenylase complex, initiating the mRNA decay reaction. Although previous studies have demonstrated the critical role of GW182 in cultured cells as well as in cell-free systems, its biological significance in living organisms remains poorly explored, especially in Drosophila melanogaster. Here, we generated gw182-null flies using the CRISPR/Cas9 system and found that, unexpectedly, they can survive until an early second-instar larval stage. Moreover, in vivo miRNA reporters can be effectively repressed in gw182-null first-instar larvae. Nevertheless, gw182-null flies have defects in the expression of chitin-related genes and the formation of the larval trachea system, preventing them from completing larval development. Our results highlight the importance of both GW182-dependent and -independent silencing mechanisms in vivo., Competing Interests: Disclosure and competing interests statement. The authors declare no competing interests. Yukihide Tomari is a member of the Advisory Editorial Board of The EMBO Journal. This has no bearing on the editorial consideration of this article for publication., (© 2024. The Author(s).)

Human activities affect natural ecosystems worldwide and can generate negative effects on insect species such as growth inhibition, developmental abnormalities and reduction of reproductive and survival rates. Our study focused on fine particulate matter, a pollutant known to cause mechanical obstruction, heavy metal intoxication, and stress in Lepidoptera larvae. Heliconius ethilla (Lepidoptera: Nymphalidae) is a Heliconiinae found in the southeastern region of Brazil. We started our study with 3255 eggs, from which 69 stage 3 larvae were randomly separated as the treatment group. The larvae were fed with Passiflora edulis leaves and the SPM was added before being offered to the treatment group in an increasing concentration according to the age of the larvae and the leaf, simulating the gradual deposition of SPM in the environment. Our results demonstrate that Sedimentable Particulate Matter negatively impacted mortality rates, pupal weight, and body size. The results strongly indicate that the presence of SPM negatively impacts the survival, development and potentially the reproductive success of H. ethilla. A reduction in the population size and a consequent decrease in the chances of long-term survival of the species can be expected., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Long-term datasets provide context and understanding of complex ecological processes, including temporal variations in species diversity and ecosystem dynamics. This dataset is comprised of body length measurements (mm) of more than 380,000 larval or juvenile fish of 30 species from five English river catchments collected almost monthly over two decades. Such information can be used to determine growth rates, future recruitment success, population structure and compliance with monitoring protocols and conservation objectives. The dataset provides a baseline for analysing the impacts of anthropogenic disturbances such as climate change, pollution and habitat degradation, and, given that many fish populations are size structured with a positive relationship between fish body length and various biological attributes such as swimming ability, survival and fecundity, it will be invaluable for investigating natural and human- induced disturbances., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

The black soldier fly, Hermetia illucens (L. 1758), is an omnivorous saprophagous insect with a high potential for valorizing organic by-products rich in carbohydrates. Among carbohydrates, H. illucens relies on soluble sugars for the growth and storage lipid synthesis. This study aimed to assess the impact of common soluble sugars on the development, survival, and fatty acid composition of H. illucens. Monosaccharides and disaccharides were individually incorporated into a chicken feed diet. Cellulose was used as a control. Larvae fed glucose, fructose, sucrose, and maltose grew faster than control larvae. In contrast, lactose exhibited anti-nutritional effects on larvae, slowing down growth and reducing final individual weight. However, all soluble sugars produced larvae fatter than the control diet. Notably, the tested sugars shaped the fatty acid profile. Maltose and sucrose increased saturated fatty acid content compared to cellulose. Conversely, lactose increased the bioaccumulation of dietary unsaturated fatty acids. This study is the first to demonstrate the effect of soluble sugars on the fatty acid profile of H. illucens larvae. Our findings highlight that the tested carbohydrates have significant effects on black soldier fly fatty acid composition and can thus determine their final applications., Competing Interests: Declarations Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

In the life cycle of the oriental fruit fly where larvae reside within fruits and adults exhibit high activity, the pupal stage occurs in the soil, closely tied to agricultural soil management. This study investigates the impact of four variables (body orientation, burial depth, soil particle size, and pH) on Bactrocera dorsalis' physiological preferences, eclosion rates, and pupal stage duration. Notably, body orientation, burial depth, soil texture (particle size), and pH affect eclosion rates. The fruit fly demonstrates a preference for a supine position during eclosion, with average eclosion rates of 97.33%, contrasting with decreases to 78.00% in a head-down position. Soil with mixed particle sizes is detrimental to pupal eclosion, reducing the eclosion rate to 56.67%. And at a pH of 9.98, eclosion rates decrease to 16.67%. These factors significantly influence eclosion rates However, these factors do not significantly alter the duration of the pupal stage, with mean changes not exceeding one day when experimental conditions are modified. These findings suggests that soil manipulations affecting these variables could reduce eclosion rates without compromising the uniformity of adult emergence. This study provides a foundation for environmentally friendly pest control practices and addresses gaps in the pupal stage research of the oriental fruit fly., Competing Interests: Declarations Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Otopheidomenidae include 30 species of parasites of insects. These mites occur in areas with warm and hot climates. The only exception is Hemipteroseius adleri, the distribution of which reaches the central European region, which is characterised by a moderate climate with below-zero temperatures in winter. In Europe, H. adleri only parasitises the red firebug (Pyrrhocoris apterus). Field observations indicate that only fertilised females of H. adleri survive winter conditions. The aim of the study therefore was to evaluate the impact of air temperature on the development and survival of H. adleri. A research model was established involving mites parasitising red firebugs in laboratory conditions at 15, 20, 25 and 30 °C. After oviposition, no egg development was possible at 15 °C. Egg development began at 20 °C, however the egg-to-adult period lasted for 13 days and larval mortality was significantly higher at this temperature than at 25 and 30 °C. At the latter tem temperatures, mites matured fastest (five and four days, respectively). The net reproductive rate (R 0 ) was 10.8, the mean generation time was (T) 14.4 (SD = 0.273), the intrinsic rate of population increase (r m ) was 0.165 (SD = 0.003), and the finite rate of population increase (λ) was 1.12. At 25 °C, H. adleri required 4.2 days to double its population. The results indicate that the thermal conditions prevailing in early spring in central Europe are highly unfavourable for the effective reproduction of H. adleri. The presence of northern populations of this mite is discussed in the context of behavioural thermoregulation mechanisms adapted by its host, P. apterus., (© 2024. The Author(s).)

Rapid technological advances and growing participation from amateur naturalists have made countless images of insects in their natural habitats available on global web portals. Despite advances in automated species identification, traits like developmental stage or health remain underexplored or manually annotated, with limited focus on automating these features. As a proof-of-concept, we developed a computer vision model utilizing the YOLOv5 algorithm to accurately detect monarch butterfly caterpillars in photographs and classify them into their five developmental stages (instars). The training data were obtained from the iNaturalist portal, and the photographs were first classified and annotated by experts to allow supervised training of models. Our best trained model demonstrates excellent performance on object detection, achieving a mean average precision score of 95% across all five instars. In terms of classification, the YOLOv5l version yielded the best performance, reaching 87% instar classification accuracy for all classes in the test set. Our approach and model show promise in developing detection and classification models for developmental stages for insects, a resource that can be used for large-scale mechanistic studies. These photos hold valuable untapped information, and we've released our annotated collection as an open dataset to support replication and expansion of our methods., (© 2024. The Author(s).)

Wireworms, the larva of click beetle (Agriotes species), are one of the most destructive pests of horticultural crops in North America, responsible for considerable economic losses in Canada. Agriotes sputator (A. sputator) species is a predominant wireworm pest attacking potato fields in Eastern Canada. However, no information about its genome-wide gene expression profile, specifically for the genes involved with development is available to date. Therefore, we generated the transcriptome profile of A. sputator during five developmental stages, including the three larval stages and adult male and female click beetle. Out of 714.7 million raw reads, de novo assembly generated 564,561 transcripts. The data were subjected to differential expression analysis using DESeq2, gene ontology, annotation, and pathway analyses. A total of 34,709 differentially expressed genes (DEGs) were significant (log 2 fold change > 2, padj < 0.05) across the developmental stages. Functional analysis of DEGs identified development signaling, metabolism, transport, cellular mechanisms, and drug metabolism (cytochrome p450) pathways. This study provides comprehensive sequence resources and potential gene differences at different developmental stages of A. sputator. These findings will represent a major step towards developing sustainable methods to control this widely distributed pest in agricultural fields., (© 2024. The Author(s).)

This study elucidated the role of DHA-modulated genes in the development and growth of Atlantic bluefin tuna (Thunnus thynnus) larvae ingesting increasing levels of DHA in their rotifer prey. The effect of feeding low, medium, and high rotifer (Brachionus rotundiformis) DHA levels (2.0, 3.6 and 10.9 mg DHA g -1 DW, respectively) was tested on 2-15 days post hatching (dph) bluefin tuna larvae. Larval DHA content markedly (P < 0.05) increased in a DHA dose-dependent manner (1.5, 3.9, 6.1 mg DHA g -1 DW larva, respectively), that was positively correlated with larval prey consumption and growth (P < 0.05). Gene ontology enrichment analyses of differentially expressed genes (DEGs) demonstrated dietary DHA significantly (P < 0.05) affected different genes and biological processes at different developmental ages. The number of DHA up-regulated DEGs was highest in 10 dph larvae (491), compared to 5 (12) and 15 dph fish (34), and were mainly involved in neural and synaptic development in the brain and spinal cord. In contrast, DHA in older 15 dph larvae elicited fewer DEGs but played critical roles over a wider range of developing organs. The emerging picture underscores the importance of DHA-modulated gene expression as a driving force in bluefin tuna larval development and growth., (© 2024. The Author(s).)

This study conducts a comprehensive analysis and comparison of Bombyx mori cuticles across different developmental stages, ranging from larval to adult, utilizing advanced solid-state NMR techniques. The primary objective is to elucidate the underlying reasons for the contrasting hardness of adult cuticles and softness of larval cuticles. Notably, PXRD analysis reveals a prominent broad peak at 19.34°, indicating the predominantly amorphous nature of both larval and adult cuticles. Analysis of 13 C CP-MAS SSNMR spectra highlights an elevated proportion of phenoxy carbon in adult cuticles (6.77%) compared to larval cuticles (1.24%). Furthermore, a distinctive resonance line at 144 ppm is exclusively observed in adult cuticles, due to catechols, suggesting potential biochemical pathway variations during development. Significant variations in the primary components of 13 C chemical shift anisotropy (CSA) tensors for aliphatic carbons of amino acids, catechols, and lipids between adult and larval cuticles indicate alterations in electronic environments. Additionally, the shorter spin-lattice relaxation time of carbon nuclei in larval cuticles compared to adult cuticles implies slower motional dynamics with enhanced degree of sclerotization in adults. By investigating the internal structure and dynamics of cuticles, this research not only contributes to biomimetic material development but also enhances our understanding of structural changes across different developmental stages of B. mori., (© 2024. The Author(s).)

Plastic waste has recently become a major global environmental concern and one of the biggest challenges has been seeking for alternative management options. Several studies have revealed the potential of several coleopteran species to degrade plastics, and this is the first research paper on plastic-degradation potential by lesser mealworms from Africa. This study evaluated the whole mitogenomic profile of the lesser mealworm to further identify the insect. The ability of the mealworm to consume Polystyrene (PS) was also evaluated alongside its associated gut microbiota diversity. Our results showed a complete circular mitochondrial genome which clustered closely to the Alphitobius genus but also suggested that our insect might be a new subspecies which require further identification. During the PS feeding trials, overall survival rates of the larvae decreased when fed a sole PS diet while PS intake was observed to increase over a 30-day period. The predominant bacteria observed in larvae fed PS diets were Kluyvera, Lactococcus, Klebsiella, Enterobacter, and Enterococcus, while Stenotrophomonas dominated the control diet. These findings demonstrated that the newly identified lesser mealworm can survive on a PS diet and has a consortium of important bacteria strongly associated with PS degradation. This work provides a better understanding of bioremediation applications, paving the way for further research into the metabolic pathways of plastic-degrading microbes and bringing hope to solving plastic waste pollution while providing high-value insect protein towards a circular economy., (© 2024. The Author(s).)

During their development, amphibians undergo various physiological processes that may affect their susceptibility to environmental pollutants. Naturally occurring fluctuations caused by developmental events are often overlooked in ecotoxicological studies. Our aim is to investigate how biomarkers of oxidative stress are modulated at different stages of larval development in the Amazonian amphibian species, Physalaemus ephippifer. The premetamorphosis, prometamorphosis and metamorphic climax stages were used to analyze total antioxidant capacity (ACAP), glutathione S-transferase (GST) activity, lipid peroxidation (LPO) levels and the expression of genes nrf2, gst, gsr (glutathione reductase) and gclc (glycine-cysteine ligase, catalytic subunit). Although there was no difference in ACAP and the genes expression among the studied stages, individuals from the premetamorphosis and prometamorphosis showed higher GST activity than ones under the climax. LPO levels were highest in individuals from the metamorphic climax. The present study suggests that the oxidative status changes during ontogeny of P. ephippifer tadpoles, especially during the metamorphic climax, the most demanding developmental phase. Variations in the redox balance at different developmental stages may lead to a divergent response to pollution. Therefore, we recommend that studies using anuran larvae as biomonitors consider possible physiological differences during ontogeny in their respective analyses., (© 2024. The Author(s).)

Several studies describe the development of the chondrocranium of vertebrates. The details in these studies vary a lot, which makes it hard to compare developmental patterns and identify evolutionary trends. Therefore, we aim to close this gap for anurans, which is the largest order of amphibians. We present here a detailed description of the chondrocranium morphology and development of Smilisca phaeota, the New Granada cross-banded tree frog. The anatomy was described for the larvae at or older than Gossner stage 31 and before ossification starts. Following this, we describe the development of the chondrocranium from Gossner stages 19-26. Early in Gossner stage 19 no precursors of any cartilages are visible, while later in that stage the mesodermal Anlage of Meckel's cartilage was observed. In the subsequent stages more and more mesodermal anlagen become identifiable, followed by chondrification, and final differentiation of the cartilage elements. We used serial sections to study all the developmental stages and additionally utilized cleared and stained specimens and CT scan data. The latter were also used for the 3D reconstruction of the chondrocranium. We previously studied several species and compared these developmental patterns with S. phaeota, revealing potentially characteristic patterns significant for Lalagobatrachia, a clade that includes over 7000 frog species. These include (1) the suprarostral alae develop before the suprarostral corpus, (2) the infrarostral cartilage chondrifies late, after the chondrification of ceratobranchial 1, and (3) the ceratohyal body is the first element to show chondrocytes and to chondrify. However, with only six species studied so far, our data only provide a basis for future studies and developing hypotheses about the ancestral developmental pattern in anurans., (© 2024. The Author(s).)

The presence of different types of larvae within the same class suggests a broad ecological diversification. A clear comparison of bivalve larval nervous systems would give a broader view on evolutionary and ecological picture of the clade in question. The present study focused on the neurodevelopment in two bivalve species with different larval types: pericalymma of Acila insignis (Bivalvia: Protobranchia) and veliger of Spisula sybillae (Bivalvia: Autobranchia). It was shown that the pioneer dorsal and ventral neurons in S. sybillae appear at the trochophore stage. Subsequently, future three paired ganglia are developed on the nerve cords in pediveliger. In the pericalymma of A. insignis, serotonin- and FMRFamide-positive cells are found in the apical organ (AO), as well as two pairs of FMRFamide positive neurons are detected on dorsal and posterior part of pericalymma. A comparative analysis showed significant differences in the larval neuromorphology between veliger and pericalymma. In contrast to the S. sybillae veliger, the nervous system of the A. insignis pericalymma is simple, likely due to its different lifestyle. The larval nervous system in the species under study has features characteristic of Lophotrochozoa and Spiralia., (© 2024. The Author(s).)

As reproduction phenologies shift with climate change, populations can experience intraspecific priority effects, wherein early hatching cohorts experience an advantage over late-hatching cohorts, resulting in altered demography. Our study objective was to identify how variation in egg hatching phenology alters intraspecific interactions in small-mouthed salamanders, Ambystoma texanum. We addressed two research questions: (Q1) How are demographic responses altered by variation in the temporal duration of hatching between cohorts, and (Q2) How does the seasonality of hatching delays affect demographic responses? We manipulated hatching phenologies of A. texanum eggs and reared larvae in outdoor mesocosms to metamorphosis. For Q1, hatching delay exhibited non-linear relationships with survival and body size, with the greatest asynchrony in cohort additions resulting in the highest mortality and largest body sizes. For Q2, hatching delay effects were stronger (i.e., survival was lower and body sizes larger) when they occurred later in the season, potentially due to temperature differences that larvae experienced. Overall, our results demonstrate that changes in intraspecific interactions due to phenological shifts can be context-dependent, depending on the strength (i.e., temporal duration) and seasonality of such processes. Identifying context-dependencies of phenological shifts will be critical for predicting changes in organismal demographics with climatic shifts., (© 2024. The Author(s).)

Marine heatwaves (MHWs) are often associated with physiological changes throughout biological communities but can also result in biomass declines that correspond with shifts in phenology. We examined the response of larval Pacific cod (Gadus macrocephalus) to MHWs in the Gulf of Alaska across seven years to evaluate the effects of MHWs on hatch phenology, size-at-age, and daily growth and identify potential regulatory mechanisms. Hatch dates were, on average, 19 days earlier since the onset of MHWs, shifting a mean of 15 days earlier per 1 ℃ increase. Size-at-capture was larger during & between MHWs but, contrary to expectations, larvae grew slower and were smaller in size-at-age. The larger size during & between MHWs can be entirely explained by older ages due to earlier hatching. Daily growth variation was well-explained by an interaction among age, temperature, and hatch date. Under cool conditions, early growth was fastest for the latest hatchers. However, this variation converged at warmer temperatures, due to faster growth of earlier hatchers. Stage-specific growth did not vary with temperature, remaining relatively similar from 4 to 8 ℃. Temperature-related demographic changes were more predictable based on phenological shifts rather than changes in growth, which could affect population productivity after MHWs., (© 2024. The Author(s).)

Drastic increases in myofiber number and size are essential to support vertebrate post-embryonic growth. However, the collective cellular behaviors that enable these increases have remained elusive. Here, we created the palmuscle myofiber tagging and tracking system for in toto monitoring of the growth and fates of ~5000 fast myofibers in developing zebrafish larvae. Through live tracking of individual myofibers within the same individuals over extended periods, we found that many larval myofibers readily dissolved during development, enabling the on-site addition of new and more myofibers. Remarkably, whole-body surveillance of multicolor-barcoded myofibers further unveiled a gradual yet extensive elimination of larval myofiber populations, resulting in near-total replacement by late juvenile stages. The subsequently emerging adult myofibers are not only long-lasting, but also morphologically and functionally distinct from the larval populations. Furthermore, we determined that the elimination-replacement process is dependent on and driven by the autophagy pathway. Altogether, we propose that the whole-body replacement of larval myofibers is an inherent yet previously unnoticed process driving organismic muscle growth during vertebrate post-embryonic development., (© 2024. The Author(s).)

Time-course transcriptome expression data were constructed for four parts of the silk gland (anterior, middle, and posterior parts of the middle silk gland, along with the posterior silk gland) in the domestic silkworm, Bombyx mori, from days 0 to 7 of the last-instar larvae. For sample preparation, silk glands were extracted from one female and one male larva every 24 hours accurately after the fourth ecdysis. The reliability of these transcriptome data was confirmed by comparing the transcripts per million (TPM) values of the silk gene and quantitative reverse transcription PCR results. Hierarchical cluster analysis results supported the reliability of transcriptome data. These data are likely to contribute to the progress in molecular biology and genetic research using B. mori, such as elucidating the mechanism underlying the massive production of silk proteins, conducting entomological research using a meta-analysis as a model for lepidopteran insect species, and exploring medical research using B. mori as a model for disease species by utilising transcriptome data., (© 2024. The Author(s).)

Marine Heatwaves (MHWs) can directly influence survival of marine fishes, particularly for early life stages, including age-0 juveniles during their residence in coastal nursery habitats. However, the ability of nurseries to support high fish densities, optimize foraging and growth, and protect against predators may be altered during MHWs. Gulf of Alaska Pacific cod (Gadus macrocephalus) larval, juvenile, and adult abundances declined dramatically following MHW events in 2014-2016 and 2019. To evaluate coastal nursery function during MHWs, we compared diet composition, recent growth, size, condition, and abundance of age-0 juveniles throughout their first summer before, during, and between MHWs. Diet shifted to larger prey during MHWs, particularly mysids, but diet did not appear to influence growth. We observed faster growth rates during MHWs, yet even when accounting for growth, we could not explain the higher body sizes observed in August during MHWs. Together with lower abundance and the near absence of small fish in the nursery by August during MHWs, these patterns highlight potential for size-selection and a reduced ability of nursery habitats to buffer against environmental variability during MHWs, with only a small number of large "super survivors" persisting through the summer., (© 2024. The Author(s).)

Bacteria induced metamorphosis observed in nearly all marine invertebrates. However, the mechanism of bacteria regulating the larvae-juvenile metamorphosis remains unknown. Here, we test the hypothesis that c-di-GMP, a ubiquitous bacterial second-messenger molecule, directly triggers the mollusc Mytilus coruscus larval metamorphosis via the stimulator of interferon genes (STING) receptor. We determined that the deletion of c-di-GMP synthesis genes resulted in reduced c-di-GMP levels and biofilm-inducing activity on larval metamorphosis, accompanied by alterations in extracellular polymeric substances. Additionally, c-di-GMP extracted from tested varying marine bacteria all exhibited inducing activity on larval metamorphosis. Simultaneously, through pharmacological and molecular experiments, we demonstrated that M. coruscus STING (McSTING) participates in larval metamorphosis by binding with c-di-GMP. Our findings reveal that new role of bacterial c-di-GMP that triggers mussel larval metamorphosis transition, and extend knowledge in the interaction of bacteria and host development in marine ecosystems., (© 2024. The Author(s).)

The beet armyworm, Spodoptera exigua (Hubner) (Lepidoptera: Noctuidae), has become a significant pest of chickpea in recent years. The polyphagous nature allows it to survive on various hosts during the off-season, creating a great menace to the crop in the following season. To assess the incidence and document the alternate hosts of S. exigua, a rapid roving survey was conducted in 11 chickpea-growing areas of Prakasam district, Andhra Pradesh, India. Additionally, the life history traits of S. exigua were studied on major alternate host plants under laboratory conditions (27 ± 1 °C and 70 ± 2% RH) to understand the survival, life expectancy and potential contribution to future populations. The results show that, among the different crops surveyed, the maximum larval incidence was noticed in maize (1.93 larvae/plant), cowpea (1.73 larvae/plant), and sunflower (1.68 larvae/plant) during the off-season. Life history studies of S. exigua showed that highest larval survival percentage was observed on chickpea (83.6%), while the lowest was on maize (44.5%). The mean developmental time for larvae was longest on maize (27.1 days) and shortest on chickpea (14.9 days). Larvae did not develop beyond the third instar when fed with chilli. The growth index statistics showed chickpea (9.2) was the most suitable host plant, whereas maize (0.9) was the least suitable host. The age-stage-specific survival rate (S xj ) varied across developmental stages, and the survival curves overlapped, indicating different growth rates among individuals. The life expectancy (e xj ) at age zero was highest on groundnut (37.06 days). The intrinsic rate of increase (r) of S. exigua was lowest on maize (0.10 ± 0.0013) and highest on chickpea (0.22 ± 0.0010). Similarly, the net reproductive rate (R 0 ) was highest on chickpea (846.39 ± 18.22) and lowest on maize (59.50 ± 2.06). The population doubled every 3.08 ± 0.011 days on chickpea compared to 7.22 ± 0.80 days on maize. The study conclusively indicates that chickpea and sunflower, primarily cultivated during the rabi season in India, are the most preferred hosts for S. exigua. In contrast, maize and cotton, mainly grown during the kharif season, are less preferred and merely support the pest's survival. Consequently, S. exigua switches hosts between different crops growing seasons, so effective management of S. exigua during the kharif season can help prevent pest outbreaks during the rabi season., (© 2024. The Author(s).)

Insect metamorphosis involves significant changes in insect internal structure and is thus a critical focus of entomological research. Investigating the morphological transformation of internal structures is vital to understanding the origins of adult insect organs. Beetles are among the most species-rich groups in insects, but the development and transformation of their internal organs have yet to be systematically documented. In this study, we have acquired a comprehensive dataset that includes 27 detailed whole-body tomographic image sets of Harmonia axyridis, spanning from the prepupal to the pupal stages. Utilizing this data, we have created intricate 3D models of key internal organs, encompassing the brain, ventral nerve cord, digestive and excretion systems, as well as the body wall muscles. These data documented the transformation process of these critical organs and correlations between the origin of adult and larval organs and can be used to enhance the understanding of holometabolous adult organ genesis and offers a valuable reference model for investigating complete metamorphosis in insects., (© 2024. The Author(s).)

Seagrass habitats play a major role in fisheries productivity through nursery functions and feeding grounds for diverse fish species. However, little is known about the seasonal distribution of fish larvae at large spatial scales in coastal East Africa. We investigated drivers of the seasonal fish larvae abundance and composition in seagrass habitats in Kenya and Tanzania. We found a high diversity of fish larvae (54 families) inhabiting seagrass habitats that differed between sites and seasons. Fish larvae abundance were highest in Kenya, particularly during the northeast monsoon season. Overall, total larval abundances per site were low, reaching less than 190 individuals/100 m 3 in Kenya and less than 40 individuals/100 m 3 in Tanzania, likely related to the low productivity and strong hydrodynamic processes in this region. Our data suggests that most of the fish spawn year-round in these tropical waters as we did not find strong seasonal patterns. All sites had a high relative abundance of larvae from demersal spawning fishes, indicating that many fish species move to coastal sites for spawning. Primary productivity and dissolved oxygen, driven by hydrodynamics conditions are positively related to fish larvae productivity both in Kenya and Tanzania. These findings indicate that the occurrence of both resident and transient fish larvae in seagrass meadows is driven by strong hydrodynamic and tidal processes that transport fish larvae across adjacent habitats., (© 2024. The Author(s).)

The mechanisms that underlie senescence are not well understood in insects. Telomeres are conserved repetitive sequences at chromosome ends that protect DNA during replication. In many vertebrates, telomeres shorten during cell division and in response to stress and are often used as a cellular marker of senescence. However, little is known about telomere dynamics across the lifespan in invertebrates. We measured telomere length in larvae, prepupae, pupae, and adults of two species of solitary bees, Osmia lignaria and Megachile rotundata. Contrary to our predictions, telomere length was longer in later developmental stages in both O. lignaria and M. rotundata. Longer telomeres occurred after emergence from diapause, which is a physiological state with increased tolerance to stress. In O. lignaria, telomeres were longer in adults when they emerged following diapause. In M. rotundata, telomeres were longer in the pupal stage and subsequent adult stage, which occurs after prepupal diapause. In both species, telomere length did not change during the 8 months of diapause. Telomere length did not differ by mass similarly across species or sex. We also did not see a difference in telomere length after adult O. lignaria were exposed to a nutritional stress, nor did length change during their adult lifespan. Taken together, these results suggest that telomere dynamics in solitary bees differ from what is commonly reported in vertebrates and suggest that insect diapause may influence telomere dynamics., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

We present a de novo transcriptome of the mosquito vector Culex pipiens, assembled by sequences of susceptible and insecticide resistant larvae. The high quality of the assembly was confirmed by TransRate and BUSCO. A mapping percentage until 94.8% was obtained by aligning contigs to Nr, SwissProt, and TrEMBL, with 27,281 sequences that simultaneously mapped on the three databases. A total of 14,966 ORFs were also functionally annotated by using the eggNOG database. Among them, we identified ORF sequences of the main gene families involved in insecticide resistance. Therefore, this resource stands as a valuable reference for further studies of differential gene expression as well as to identify genes of interest for genetic-based control tools., (© 2024. The Author(s).)

One of the major functions of the larval salivary glands (SGs) of many Drosophila species is to produce a massive secretion during puparium formation. This so-called proteinaceous glue is exocytosed into the centrally located lumen, and subsequently expectorated, serving as an adhesive to attach the puparial case to a solid substrate during metamorphosis. Although this was first described almost 70 years ago, a detailed description of the morphology and mechanical properties of the glue is largely missing. Its main known physical property is that it is released as a watery liquid that quickly hardens into a solid cement. Here, we provide a detailed morphological and topological analysis of the solidified glue. We demonstrated that it forms a distinctive enamel-like plaque that is composed of a central fingerprint surrounded by a cascade of laterally layered terraces. The solidifying glue rapidly produces crystals of KCl on these alluvial-like terraces. Since the properties of the glue affect the adhesion of the puparium to its substrate, and so can influence the success of metamorphosis, we evaluated over 80 different materials for their ability to adhere to the glue to determine which properties favor strong adhesion. We found that the alkaline Sgs-glue adheres strongly to wettable and positively charged surfaces but not to neutral or negatively charged and hydrophobic surfaces. Puparia formed on unfavored materials can be removed easily without leaving fingerprints or cascading terraces. For successful adhesion of the Sgs-glue, the material surface must display a specific type of triboelectric charge. Interestingly, the expectorated glue can move upwards against gravity on the surface of freshly formed puparia via specific, unique and novel anatomical structures present in the puparial's lateral abdominal segments that we have named bidentia., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Eicosanoids are synthesized from phospholipids by the catalytic activity of phospholipase A 2 (PLA 2 ). Even though several PLA 2 s are encoded in the genome of different insect species, their physiological functions are not clearly discriminated. This study identified four PLA 2 genes encoded in the western flower thrips, Frankliniella occidentalis. Two PLA 2 s (Fo-PLA 2 C and Fo-PLA 2 D) are predicted to be secretory while the other two PLA 2 s (Fo-PLA 2 A and Fo-PLA 2 B) are intracellular. All four PLA 2 genes were expressed in all developmental stages, of which Fo-PLA 2 B and Fo-PLA 2 C were highly expressed in larvae while Fo-PLA 2 A and Fo-PLA 2 D were highly expressed in adults. Their expressions in different tissues were also detected by fluorescence in situ hybridization. All four PLA 2 s were detected in the larval and adult intestines and the ovary. Feeding double-stranded RNAs specific to the PLA 2 genes specifically suppressed the target transcript levels. Individual RNA interference (RNAi) treatments led to significant developmental retardation, especially in the treatments specific to Fo-PLA 2 B and Fo-PLA 2 D. The RNAi treatments also showed that Fo-PLA 2 B and Fo-PLA 2 C expressions were required for the induction of immune-associated genes, while Fo-PLA 2 A and Fo-PLA 2 D expressions were required for ovary development. These results suggest that four PLA 2 s are associated with different physiological processes by their unique catalytic activities and expression patterns., (© 2024. The Author(s).)

The development of paired appendages was a key innovation during evolution and facilitated the aquatic to terrestrial transition of vertebrates. Largely derived from the lateral plate mesoderm (LPM), one hypothesis for the evolution of paired fins invokes derivation from unpaired median fins via a pair of lateral fin folds located between pectoral and pelvic fin territories 1 . Whilst unpaired and paired fins exhibit similar structural and molecular characteristics, no definitive evidence exists for paired lateral fin folds in larvae or adults of any extant or extinct species. As unpaired fin core components are regarded as exclusively derived from paraxial mesoderm, any transition presumes both co-option of a fin developmental programme to the LPM and bilateral duplication 2 . Here, we identify that the larval zebrafish unpaired pre-anal fin fold (PAFF) is derived from the LPM and thus may represent a developmental intermediate between median and paired fins. We trace the contribution of LPM to the PAFF in both cyclostomes and gnathostomes, supporting the notion that this is an ancient trait of vertebrates. Finally, we observe that the PAFF can be bifurcated by increasing bone morphogenetic protein signalling, generating LPM-derived paired fin folds. Our work provides evidence that lateral fin folds may have existed as embryonic anlage for elaboration to paired fins., (© 2023. The Author(s).)

Indirect development with an intermediate larva exists in all major animal lineages 1 , which makes larvae central to most scenarios of animal evolution 2-11 . Yet how larvae evolved remains disputed. Here we show that temporal shifts (that is, heterochronies) in trunk formation underpin the diversification of larvae and bilaterian life cycles. We performed chromosome-scale genome sequencing in the annelid Owenia fusiformis with transcriptomic and epigenomic profiling during the life cycles of this and two other annelids. We found that trunk development is deferred to pre-metamorphic stages in the feeding larva of O. fusiformis but starts after gastrulation in the non-feeding larva with gradual metamorphosis of Capitella teleta and the direct developing embryo of Dimorphilus gyrociliatus. Accordingly, the embryos of O. fusiformis develop first into an enlarged anterior domain that forms larval tissues and the adult head 12 . Notably, this also occurs in the so-called 'head larvae' of other bilaterians 13-17 , with which the O. fusiformis larva shows extensive transcriptomic similarities. Together, our findings suggest that the temporal decoupling of head and trunk formation, as maximally observed in head larvae, facilitated larval evolution in Bilateria. This diverges from prevailing scenarios that propose either co-option 9,10 or innovation 11 of gene regulatory programmes to explain larva and adult origins., (© 2023. The Author(s).)

Drosophila melanogaster tumor models are growing in popularity, driven by the high degree of genetic as well as functional conservation to humans. The most common method to measure the effects of a tumor on distant organs of a human cancer patient is to use computed tomography (CT), often used in diagnosing cachexia, a debilitating cancer-induced syndrome most visibly characterized by loss of muscle mass. Successful application of high resolution micro-CT scanning of D. melanogaster was recently reported and we here present the segmentation of all visible larval organs at several stages of tumor development. We previously showed the strong expected reduction in muscle mass as the tumor develops, and we here report a surprisingly strong reduction also in gut and Malpighian tubules (kidney) volume. Time-point of tumor development was found to have a stronger correlation to cachectic organ volume loss than tumor volume, giving support to the previously proposed idea that tumor size does not directly determine degree of cachexia., (© 2022. The Author(s).)

Xenopus laevis tadpoles possess high regenerative ability and can regenerate functional tails after amputation. An early event in regeneration is the induction of undifferentiated cells that form the regenerated tail. We previously reported that interleukin-11 (il11) is upregulated immediately after tail amputation to induce undifferentiated cells of different cell lineages, indicating a key role of il11 in initiating tail regeneration. As Il11 is a secretory factor, Il11 receptor-expressing cells are thought to mediate its function. X. laevis has a gene annotated as interleukin 11 receptor subunit alpha on chromosome 1L (il11ra.L), a putative subunit of the Il11 receptor complex, but its function has not been investigated. Here, we show that nuclear localization of phosphorylated Stat3 induced by Il11 is abolished in il11ra.L knocked-out culture cells, strongly suggesting that il11ra.L encodes an Il11 receptor component. Moreover, knockdown of il11ra.L impaired tadpole tail regeneration, suggesting its indispensable role in tail regeneration. We also provide a model showing that Il11 functions via il11ra.L-expressing cells in a non-cell autonomous manner. These results highlight the importance of il11ra.L-expressing cells in tail regeneration., (© 2022. The Author(s).)

Competition within and among species can play a key role in structuring the assemblages of anuran tadpoles. Previous studies have reported that tadpoles of the invasive cane toad (Rhinella marina) are more strongly disadvantaged by the presence of native frog tadpoles than by the same number of conspecific toad tadpoles. That effect might arise from a lack of coevolution of the invasive toad with its competitors; and/or from a generalized superiority of frog tadpoles over toad tadpoles. To clarify those possibilities, we conducted experimental trials using the larvae of a native rather than invasive toad (Bufo japonicus formosus in Japan) exposed to larvae of native anurans (the sympatric frogs Rana japonica and Rana ornativentris and the parapatric toad Bufo japonicus japonicus). In intraspecific competition trials, higher densities of B. j. formosus prolonged the larval period and reduced size at metamorphosis, but did not affect survival. In interspecific competition trials, the effects of the other anuran species on B. j. formosus were similar to the effects of the same number of conspecific larvae. This similarity in impact of interspecific versus intraspecific competition argues against any overall competitive superiority of frog larvae over toad larvae. Instead, the vulnerability of larval cane toads to frog tadpoles may result from a lack of coevolutionary history., (© 2022. The Author(s).)

Protease inhibitors (PIs) are important biotechnological tools of interest in agriculture. Usually they are the first proteins to be activated in plant-induced resistance against pathogens. Therefore, the aim of this study was to characterize a Theobroma cacao trypsin inhibitor called TcTI. The ORF has 740 bp encoding a protein with 219 amino acids, molecular weight of approximately 23 kDa. rTcTI was expressed in the soluble fraction of Escherichia coli strain Rosetta [DE3]. The purified His-Tag rTcTI showed inhibitory activity against commercial porcine trypsin. The kinetic model demonstrated that rTcTI is a competitive inhibitor, with a Ki value of 4.08 × 10 -7  mol L -1 . The thermostability analysis of rTcTI showed that 100% inhibitory activity was retained up to 60 °C and that at 70-80 °C, inhibitory activity remained above 50%. Circular dichroism analysis indicated that the protein is rich in loop structures and β-conformations. Furthermore, in vivo assays against Helicoverpa armigera larvae were also performed with rTcTI in 0.1 mg mL -1 spray solutions on leaf surfaces, which reduced larval growth by 70% compared to the control treatment. Trials with cocoa plants infected with Mp showed a greater accumulation of TcTI in resistant varieties of T. cacao, so this regulation may be associated with different isoforms of TcTI. This inhibitor has biochemical characteristics suitable for biotechnological applications as well as in resistance studies of T. cacao and other crops., (© 2022. The Author(s).)

A germ-free rearing system is a crucial method for host-microbiota interactions using Nasonia as a model system. The previous rearing media in 2012 introduced toxic factors like bleach and antibiotics, required significant effort and volume of media preparation, and the rearing protocols in 2012 and 2016 often resulted in embryos, larvae, and enclosing pupae drowning, underfed, or desiccating. In this work, we optimize the germ-free rearing media that excludes the toxic factors and provide a substrate for the developing animals to have constant access to media without the risk of drowning or desiccation. The new process resulted in an increase in full maturation of larvae to adults from 33 to 65%, with no effect on the rate of growth or final adult size. This significantly improves the applicability of germ-free rearing of Nasonia and potentially other parasitoids., (© 2022. The Author(s).)

The ability to cryopreserve mosquitoes would revolutionize work on these vectors of major human infectious diseases by conserving stocks, new isolates, lab-bred strains, and transgenic lines that currently require continuous life cycle maintenance. Efforts over several decades to develop a method for cryopreservation have, until now, been fruitless: we describe here a method for the cryopreservation of Anopheles stephensi embryos yielding hatch rates of ~ 25%, stable for > 5 years. Hatched larvae developed into fertile, fecund adults and blood-fed females, produced fully viable second generation eggs, that could be infected with Plasmodium falciparum at high intensities. The key components of the cryopreservation method are: embryos at 15-30 min post oviposition, two incubation steps in 100% deuterated methanol at - 7 °C and - 14.5 °C, and rapid cooling. Eggs are recovered by rapid warming with concomitant dilution of cryoprotectant. Eggs of genetically modified A. stephensi and of A. gambiae were also successfully cryopreserved. This enabling methodology will allow long-term conservation of mosquitoes as well as acceleration of genetic studies and facilitation of mass storage of anopheline mosquitoes for release programs., (© 2022. The Author(s).)

Regional optimisation of malaria vector control approaches requires detailed understanding both of the species composition of Anopheles mosquito communities, and how they vary over spatial and temporal scales. Knowledge of vector community dynamics is particularly important in settings where ecohydrological conditions fluctuate seasonally and inter-annually, such as the Barotse floodplain of the upper Zambezi river. DNA barcoding of anopheline larvae sampled in the 2019 wet season revealed the predominance of secondary vector species, with An. coustani comprising > 80% of sampled larvae and distributed ubiquitously across all ecological zones. Extensive larval sampling, plus a smaller survey of adult mosquitoes, identified geographic clusters of primary vectors, but represented only 2% of anopheline larvae. Comparisons with larval surveys in 2017/2018 and a contemporaneous independent 5-year dataset from adult trapping corroborated this paucity of primary vectors across years, and the consistent numerical dominance of An. coustani and other secondary vectors in both dry and wet seasons, despite substantial inter-annual variation in hydrological conditions. This marked temporal consistency of spatial distribution and anopheline community composition presents an opportunity to target predominant secondary vectors outdoors. Larval source management should be considered, alongside prevalent indoor-based approaches, amongst a diversification of vector control approaches to more effectively combat residual malaria transmission., (© 2022. The Author(s).)

Ineffectiveness of carbapenems against multidrug resistant pathogens led to the increased use of colistin (polymyxin E) as a last resort antibiotic. A gene belonging to the DedA family encoding conserved membrane proteins was previously identified by screening a transposon library of K. pneumoniae ST258 for sensitivity to colistin. We have renamed this gene dkcA (dedA of Klebsiella required for colistin resistance). DedA family proteins are likely membrane transporters required for viability of Escherichia coli and Burkholderia spp. at alkaline pH and for resistance to colistin in a number of bacterial species. Colistin resistance is often conferred via modification of the lipid A component of bacterial lipopolysaccharide with aminoarabinose (Ara4N) and/or phosphoethanolamine. Mass spectrometry analysis of lipid A of the ∆dkcA mutant shows a near absence of Ara4N in the lipid A, suggesting a requirement for DkcA for lipid A modification with Ara4N. Mutation of K. pneumoniae dkcA resulted in a reduction of the colistin minimal inhibitory concentration to approximately what is found with a ΔarnT strain. We also identify a requirement of DkcA for colistin resistance that is independent of lipid A modification, instead requiring maintenance of optimal membrane potential. K. pneumoniae ΔdkcA displays reduced virulence in Galleria mellonella suggesting colistin sensitivity can cause loss of virulence., (© 2021. The Author(s).)

The current study describes the effects of sub-lethal concentrations and constituent compounds (citral and geranyl acetate) of Cymbopogon flexuosus essential oil (EO) on the development of Aedes aegypti. We treated eggs with 6, 18, or 30 mg L -1 and larvae with 3 or 6 mg L -1 of EO and its major compounds (citral and geranyl acetate). Citral and geranyl acetate were evaluated at 18, 30, and 42 mg L -1 and compared with commercial growth inhibitors (diflubenzuron and methoprene). We measured larval head diameter, siphon length, and larval length. Finally, we examined concentrations of molt hormone (MH) and juvenile hormone III (JH III) using high-performance liquid chromatography coupled to mass spectrometry. All geranyl acetate concentrations decreased egg hatching, while EO altered molting among larval instars and between larvae and pupae, with an increase in the larval length (3 mg L -1 : 6 ± 0.0 mm; 6 mg L -1 : 6 ± 0.7 mm) and head width (3 mg L -1 : 0.8 ± 0 mm; 6 mg L -1 : 0.8 ± 0.0 mm) compared with the control group. We did not detect chromatographic signals of MH and JH III in larvae treated with C. flexuosus EO or their major compounds. The sub-lethal concentrations C. flexuosus EO caused a similar effect to diflubenzuron, namely decreased hormone concentrations, an extended larval period, and death., (© 2021. The Author(s).)

The non-persistent impact of biocontrol agents can be revealed for pest control when associated entomopathogenic fungi (EPFs) negatively affect the natural enemies. In this assay, impacts of Beauvaria bassiana (Balsamo) Vuillemin, and Metarhizium anisopliae (Metschnikoff) Sorokin were studied for their compatibility or side effects on life table parameters of an important generalist predator, Coccinella septempunctata L. The results indicated non-significant impacts of both EPFs on life table parameters of C. septempunctata. The development time (egg-adult) was not significantly different in control (69.79 days) and EPFs treated C. septempunctata (69.35-80.07 days). Both fungi did not induce any significant changes in the fecundity, adult pre-oviposition period (APOP), total preoviposition period (TPOP), and mean generation time (T) as compared to control treatment. Similarly, no difference in fecundity rate of C. septempunctata was observed after EPFs treatment (287.7-288.5) compared to control (290.0). The highest net reproductive rate (R 0 ) occurred in control (87.05 offspring individual -1 ) and M. anisopliae (86.31 offspring individual -1 ) as compared to B. bassiana treated beetles (76.97 offspring individual -1 ). The age-specific fecundity curves indicated that the C. septempunctata had a similar fecundity rate in both EPFs treatments and control. This study demonstrates no significant side effects of B. bassiana and M. anispoliae on the performance and biology of C. septempunctata. Considering the compatibility of both EPFs with C. septempunctata, their combinations can be recommended in various integrated pest management programs., (© 2021. The Author(s).)

Culex pipiens pallens is an important vector of lymphatic filariasis and epidemic encephalitis. Mosquito control is the main strategy used for the prevention of mosquito-borne diseases. Bacillus thuringiensis israelensis (Bti) is an entomopathogenic bacterium widely used in mosquito control. In this study, we profiled the microbiota and transcriptional response of the larvae of Cx. pipiens pallens exposed to different concentrations of Bti. The results demonstrated that Bti induced a significant effect on both the microbiota and gene expression of Cx. pipiens pallens. Compared to the control group, the predominant bacteria changed from Actinobacteria to Firmicutes, and with increase in the concentration of Bti, the abundance of Actinobacteria was gradually reduced. Similar changes were also detected at the genus level, where Bacillus replaced Microbacterium, becoming the predominant genus in Bti-exposed groups. Furthermore, alpha diversity analysis indicated that Bti exposure changed the diversity of the microbota, possibly because the dysbiosis caused by the Bti infection inhibits some bacteria and provides opportunities to other opportunistic taxa. Pathway analysis revealed significant enhancement for processes associated with sphingolipid metabolism, glutathione metabolism and glycerophospholipid metabolism between all Bti-exposed groups and control group. Additionally, genes associated with the Toll and Imd signaling pathway were found to be notably upregulated. Bti infection significantly changed the bacterial community of larvae of Cx. pipiens pallens., (© 2021. The Author(s).)

The leafminer Liriomyza trifolii is one of the major insects that affect Phaseolus vulgaris production worldwide. Novel and safe biobased stimulator compound (BSTC) with micronutrient-amino acid chelated compounds was developed from natural compounds and was used for foliar spray of P. vulgaris. Treated plants showed significantly increased in quality and productivity as well as significant reduction in leafminer infestation by close the tunnel end resulting in larvae suffocation and death. BSTC contains chemical composition that has important function in inducing immunity and resistance against insects, enhance plant growth and production. Also, HPLC showed that the assembled BSTC is rich in nucleobases than yeast extract (> 56 fold). Aminochelation zinc enhanced the rate of absorption of nutrient compounds and could participate in safe biofortification strategy. The expression of plant defense related genes under BSTC treatment revealed strong correlations between the transcription rates of defense related genes. Based on binding energies and interacting residues of six vital insect proteins, the best-docked complexes was obtained with disodium 5'-inosinate, delphinidin 3-glucoside and hyperoside. Obtained findings indicate that the foliar application of BSTC can enhance plant growth and productivity, uptake of important elements, expression of defense related genes and inhibit insect essential genes., (© 2021. The Author(s).)

Larval metamorphosis in bivalves is a key event for the larva-to-juvenile transformation. Previously we have identified a thyroid hormone receptor (TR) gene that is crucial for larvae to acquire "competence" for the metamorphic transition in the mussel Mytilus courscus (Mc). The mechanisms of thyroid signaling in bivalves are still largely unknown. In the present study, we molecularly characterized the full-length of two iodothyronine deiodinase genes (McDx and McDy). Phylogenetic analysis revealed that deiodinases of molluscs (McDy, CgDx and CgDy) and vertebrates (D2 and D3) shared a node representing an immediate common ancestor, which resembled vertebrates D1 and might suggest that McDy acquired specialized function from vertebrates D1. Anti-thyroid compounds, methimazole (MMI) and propylthiouracil (PTU), were used to investigate their effects on larval metamorphosis and juvenile development in M. coruscus. Both MMI and PTU significantly reduced larval metamorphosis in response to the metamorphosis inducer epinephrine. MMI led to shell growth retardation in a concentration-dependent manner in juveniles of M. coruscus after 4 weeks of exposure, whereas PTU had no effect on juvenile growth. It is hypothesized that exposure to MMI and PTU reduced the ability of pediveliger larvae for the metamorphic transition to respond to the inducer. The effect of MMI and PTU on larval metamorphosis and development is most likely through a hormonal signal in the mussel M. coruscus, with the implications for exploring the origins and evolution of metamorphosis., (© 2021. The Author(s).)

Animal development relies on a sequence of specific stages that allow the formation of adult structures with a determined size. In general, juvenile stages are dedicated mainly to growth, whereas last stages are devoted predominantly to the maturation of adult structures. In holometabolous insects, metamorphosis marks the end of the growth period as the animals stops feeding and initiate the final differentiation of the tissues. This transition is controlled by the steroid hormone ecdysone produced in the prothoracic gland. In Drosophila melanogaster different signals have been shown to regulate the production of ecdysone, such as PTTH/Torso, TGFß and Egfr signaling. However, to which extent the roles of these signals are conserved remains unknown. Here, we study the role of Egfr signaling in post-embryonic development of the basal holometabolous beetle Tribolium castaneum. We show that Tc-Egfr and Tc-pointed are required to induced a proper larval-pupal transition through the control of the expression of ecdysone biosynthetic genes. Furthermore, we identified an additional Tc-Egfr ligand in the Tribolium genome, the neuregulin-like protein Tc-Vein (Tc-Vn), which contributes to induce larval-pupal transition together with Tc-Spitz (Tc-Spi). Interestingly, we found that in addition to the redundant role in the control of pupa formation, each ligand possesses different functions in organ morphogenesis. Whereas Tc-Spi acts as the main ligand in urogomphi and gin traps, Tc-Vn is required in wings and elytra. Altogether, our findings show that in Tribolium, post-embryonic Tc-Egfr signaling activation depends on the presence of two ligands and that its role in metamorphic transition is conserved in holometabolous insects., (© 2021. The Author(s).)