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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).)

Organisms produce antimicrobial peptides (AMPs) either in response to infection (induced) or continuously (constitutively) to combat microbes encountered during normal trophic activities and/or through pathogenic infections. The expression of AMPs is tightly regulated, often with specificity to particular tissues or developmental stages. As a result, AMPs face varying selective pressures based on the microbes the organism's tissue or developmental stage encounters. Here, we analyzed the evolution and developmental-specific expression of Defensins, which are ancient AMPs in insects, in the stable fly (Stomoxys calcitrans). Stable fly larvae inhibit microbe-rich environments, whereas adult flies, as blood-feeders, experience comparatively fewer encounters with diverse microbial communities. Using existing RNA-seq datasets, we identified six Defensins that were only expressed in larvae (larval Defensins) and five that were not expressed in larvae (non-larval Defensins). Each of the non-larval Defensins was expressed in at least one adult tissue sample. Half of the larval Defensins were induced by mating or feeding in adults, and all three of the induced Defensins were located downstream of canonical binding sites for an Imd transcription factor involved in the highly conserved NF-κB signaling that regulates induction of AMPs. The larval and non-larval Defensins were located in distinct genomic regions, and the amino acid sequences of the larval Defensins formed a monophyletic clade. There were more amino acid substitutions across non-larval Defensins, with multiple genes losing a highly conserved furin cleavage site thought to be required for the removal of the amino terminus from the mature Defensin domain. However, larval Defensins had a higher proportion of radical amino acid substitutions, altering amino acid size and polarity. Our results reveal insights into the developmental stage-specific regulation of AMPs, and they suggest different regulatory regimes impose unique selection pressures on AMPs, possibly as a result of variation in exposure to microbial communities across development., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology.)

Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) is a globally devastating pest and has evolved with varying levels of resistance to a broader spectrum of insecticides. CarEs are major enzymes involved in insecticide detoxification and metabolic resistance. Four CarE genes were identified and cloned from H. armigera , and the expression pattern analyses indicated that they were predominantly expressed in the detoxifying tissues and larval feeding periods. The insecticide inductive assays further suggested that these CarE genes with expressions were significantly upregulated after beta -cypermethrin, lambda -cyhalothrin, malathion, and chlorpyrifos exposures. The purified recombinant proteins of both CarE016B and CarE016C by bacterial expression exhibited significantly higher catalytic efficiency toward α-naphthyl acetate and also showed relatively stronger binding with both β-cypermethrin and λ-cyhalothrin compared to the CarE006C and CarE015A. In vitro metabolism assay with HPLC suggests that CarE016B and CarE016C have the ability to metabolize β-cypermethrin, λ-cyhalothrin, and malathion with varying hydrolase activities. GC-MS/MS analysis identified 3-phenoxybenzaldehyde (3-PBAld) as the metabolite of both β-cypermethrin and λ-cyhalothrin. Homology modeling and molecular docking analyses further indicate that these three types of insecticides could be anchored into the active pocket of both CarEs. Collectively, these results demonstrate that both CarE016B and CarE016C play a critical role in the detoxification of pyrethroid and organophosphate insecticides in H. armigera . This study provides the foundations for a comprehensive understanding of the role of the CarEs family in insecticide detoxification and resistance in H. armigera .

The widespread use of benzoylurea insecticides (BUs) has led to significant resistance issues in various agricultural pests. Previous studies have demonstrated that the overexpression of sigma glutathione S-transferase 1 (PxGSTs1) can confer resistance to novaluron in Plutella xylostella ; however, the underlying molecular mechanism remains unclear. This study investigates the role of glutathione S-transferase PxGSTs1 in mediating resistance to BUs in P. xylostella . Using a combination of RNA interference and transgenic Drosophila models, we demonstrated that the overexpression of PxGSTs1 significantly contributes to the resistance against BUs. Functional assays revealed that PxGSTs1 binds to these insecticides with varying affinities. Structural analysis through homology modeling and molecular docking identified the importance of hydrogen bonding and pi-pi stacking in resistance mechanisms. Site-directed mutagenesis confirmed the critical role of Ser65 and Tyr97 in these interactions. Our findings provide a molecular basis for the development of novel BUs and inform strategies for managing BU resistance in P. xylostella .

Black soldier fly larvae (BSFL) have attracted attention due to their ability to upcycle various biological side streams into valuable biomass, such as proteins, lipids, and chitin. In this study, we investigated the impact of high-fiber diets on larval growth performance and the shift of microbes in the gut. We tested empty fruit bunches (EFB), potato pulp (PP), and cottonseed press cake (CPC), with chicken feed (CF) used as a control diet. We found that larvae reared on the EFB, PP, and CPC were smaller than control larvae at the end of development due to the low nutritional value of the diets. However, survival rates of more than 90% were observed regardless of the diet. We used a cultivation-dependent approach to analyze the microbial community in the gut of BSFL, isolated, and identified a total of 329 bacterial strains. Bacillaceae were most frequently isolated from larvae reared on the high-fiber EFB diet. These isolates were predicted to degrade cellulose in silico and this was subsequently confirmed in vitro using the Congo Red assay. Whereas the members of Enterobacteriaceae and Morganellaceae were mostly found in guts of larvae reared on the high-protein diets CPC and CF. We conclude that the gut microbiome plays a crucial role in the digestion of fiber-rich plant organic material, thereby enabling the BSFL to successfully complete their life cycle also on substrates with low nutritional value. As a result, BSFL convert industrial side streams into valuable biomass, reducing waste and promoting sustainability., Importance: Organic side streams from various industries pose a challenge to the environment. They are often present in huge amounts and are mostly discarded, incinerated, used for biogas production, or as feed for ruminant animals. Many plant-based side streams contain difficult-to-digest fiber as well as anti-nutritional or even insecticidal compounds that could harm the animals. These challenges can be addressed using black soldier fly larvae, which are known to degrade various organic substrates and convert them into valuable biomass. This will help mitigate agro-industrial side streams via efficient waste management and will contribute to the more economical and sustainable farming of insects., Competing Interests: The authors declare no conflict of interest.

Climate change destabilizes the symbiosis between corals and Symbiodiniaceae. The effects of ocean acidification and warming on critical aspects of coral survical such as symbiotic interactions (i.e., carbon and nitrogen assimilation and exchange) during the planula larval stage remain understudied. By combining physiological and stable isotope techniques, here we show that photosynthesis and carbon and nitrogen assimilation (H 13 CO 3 - and 15 NH 4 + ) in Pocillopora damicornis coral larvae is enhanced under acidification (1000 µatm) and elevated temperature (32 °C). Larvae maintain high survival and settlement rates under these treatment conditions with no observed decline in symbiont densities or signs of bleaching. Acidification and elevated temperature both enhance the net and gross photosynthesis of Symbiodiniaceae. This enhances light respiration and elevates C:N ratios within the holobiont. The increased carbon availability is primarily reflected in the 13 C enrichment of the host, indicating a greater contribution of the algal symbionts to the host metabolism. We propose that this enhanced mutualistic symbiotic nutrient cycling may bolster coral larvae's resistance to future ocean conditions. This research broadens our understanding of the early life stages of corals by emphasizing the significance of symbiotic interactions beyond those of adult corals., Competing Interests: Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Background: Larval source management (LSM) is re-emerging as a critical malaria intervention to address challenges associated with core vector control tools, such as insecticide-treated nets (ITNs), and to accelerate progress towards elimination. Presently, LSM is not widely used in rural settings and is instead more commonly applied in urban and arid settings. A systematic entomological assessment was conducted in rural communities of southeastern Tanzania, where insecticide-treated nets (ITNs) are widely used, to explore opportunities for deploying LSM to improve malaria control., Methods: Aquatic habitat surveys were conducted in 2022 and 2023 to understand habitat usage by different mosquito vectors, covering five villages during the rainy season and seven villages during the dry season. Additionally, samples of adult mosquitoes were collected to assess the role of various Anopheles species in malaria transmission in the area, and to explore opportunities for species sanitation using targeted LSM., Results: Adult mosquito surveys showed that in this area, the total entomological inoculation rates (EIR) for indoor collections were 20.1 and 6.5 infectious bites per person per year for outdoors. Anopheles funestus and Anopheles arabiensis were the only Anopheles vectors identified. Anopheles funestus was responsible for over 97.6% of the malaria transmission indoors and 95.4% outdoors. The concurrent larval surveys found that habitats with late instar An. arabiensis and An. funestus comprised only a small subset of 11.2%-16.5% of all water bodies in the rainy season, and 9.7%-15.2% in the dry season. In terms of size, these habitats covered 66.4%-68.2% of the total habitat areas in the wet season, reducing to 33.9%-40.6% in the dry season. From the rainy season to the dry season, the surface area of habitats occupied by An. arabiensis and An. funestus decreased by 92.0% to 97.5%, while the number of habitats occupied by An. arabiensis and An. funestus decreased by 38.0% to 57.3%. Anopheles funestus preferred large, permanent habitats with clear water and vegetation year-round, while An. arabiensis showed contrasting seasonal preferences, favouring sunlit still waters in the rainy season and larger, opaque habitats in the dry season., Conclusion: These findings suggest that An. funestus, which is the dominant malaria vector in the area, mediating over 95% of malaria transmission, preferentially occupies only a small subset of uniquely identifiable aquatic habitats in both wet and dry seasons. This presents an opportunity to expand LSM in rural settings by carefully targeting An. funestus habitats, which might be effective and logistically feasible as a complementary approach alongside existing interventions. Further research should assess the impact of targeted LSM for species sanitation compared to blanket LSM., Competing Interests: Declarations Ethics approval and consent to participate Ethics approvals for this study were obtained from the Institutional Review Board of Ifakara Health Institute (Ref no: IHI/IRB/No: 32-2021) and the Medical Research Coordinating Committee (MRCC) at the National Institute for Medical Research (Ref no: NIMR/HQ/R.8a/Vol. IX/3761). Consent for publication Permission to publish was granted by The National Institute for Medical Research (NIMR), Tanzania, Ref No. BD.242/437/01C/11. Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

The objective of the study was to investigate how substituting fish meal (FM) with mealworm larval (Tenebrio molitor L.) meal (TM) in the diet affects the growth performance, meat quality, nutrient digestibility, and carcass of broiler chicks. A total of one hundred chicks were divided into five groups, each containing twenty birds. The fish meal was replaced with mealworm meal at different levels of 25% (TM25), 50% (TM50), 75% (TM75), and 100% (TM100) in the diets. At the end of the study, replacing different levels the fish meal with mealworms linearly suppressed nutrient digestion, growth, carcass parameters, and water holding capacity and cooking loss. However, the best energy efficiency ratios were obtained in the TM100 group. Consequently, it was concluded that replacement of FM with mealworm up to 25% had no adverse effect on growth performance, carcass parameters, meat quality and nutrient utilization efficiency of broiler chickens., Competing Interests: Declarations Ethics approval All experimental procedures using birds were conducted following the European Guidelines for Care and Use of Animals for Research Purpose, and they were approved by the Local Ethics Committee for Animal Testing of the Aydın Adnan Menderes University (64583101/2021/027). Conflict of interest The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

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).)

Biological treatment of food waste (FW) by black soldier fly larvae (BSFL) is considered as an effective management strategy. The composition and concentrations of nutrients in FW change during its storage and transport period, which potentially affect the FW conversion and BSFL growth. The present study systematically investigated the effect of different storage times (i.e., 0-15 d) on FW characteristics and its substantial influence on the BSFL growth. Results showed that the highest larvae weight of 282 mg and the shortest growth time of 14 days were achieved at the group of FW stored for 15 days, but shorter storage time (i.e., 2-7 d) had adverse effect on BSFL growth. Short storage time (i.e., 2-4 d) improved protein content of BSFL biomass and prolonged storage time (i.e., 7-10 d) led to the accumulation of fat content. The changes of substrate characteristics and indigenous microorganisms via FW storage time were the main reasons for BSFL growth difference. Lactic acid (LA) accumulation (i.e., 19.84 g/L) in FW storage for 7 days significantly limited the BSFL growth, leading to lowest larvae weight. Both the substrate and BSFL gut contained same bacterial communities (e.g., Klebsiella and Proteus), which exhibited similar change trend with the prolonged storage time. The transfer of Clostridioides from substrate to BSFL gut promoted nutrients digestion and intestinal flora balance with the FW stored for 15 days. Pathogens (e.g., Acinetobacter) in BSFL gut feeding with FW storage time of 7 days led to the decreased digestive function, consistent with the lowest larvae weight. Overall, shorter storage time (i.e., 2-7 d) inhibited the BSFL digestive function and growth performance, while the balance of the substrate nutrients and intestinal flora promoted the BSFL growth when using the FW stored for 15 days., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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).)

Forensic entomology plays a crucial role in estimating the minimum postmortem interval through the study of insect larvae found at crime scenes. The precision of this estimation relies on various biotic and abiotic elements that simultaneously influence insect growth and development, encompassing factors such as temperature, humidity, photoperiod, diet, and the existence of xenobiotics in decomposing tissues. Despite numerous studies on the influence of these factors, including the impact of xenobiotics, there are currently no robust tools available for making corrections to this estimation considering concurrently all variables. In an attempt to propose an exploratory and descriptive statistical model to analyze the simultaneous effect and interaction of different variables on larval growth, this study aimed to compare the effect of malathion on the growth of Megaselia scalaris (Loew, 1866) (Diptera: Phoridae) raised in malathion-spiked porcine muscle, under controlled and uncontrolled temperature and humidity conditions (environmental conditions). Larvae were also reared using various growth media. A split-plot design that combined crossed and nested factors was employed; 2 linear mixed models were developed to assess the relationships between the variables. The model provides valuable insights into the complex interactions among xenobiotics, growth media, and environmental conditions in the size and development of M. scalaris., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America.)

Insecticide formulations with safer environmental profiles and limited off-target effects are desirable to manage medical and veterinary pests. Silver nanoparticles are insecticidal against mosquitos, nonbiting midges, and other insects. The biting midge, Culicoides sonorensis Wirth and Jones, is a vector of agriculturally important pathogens in much of the United States, and this study aimed to examine the insecticidal properties of silver nanoparticles in larvae of this species. Mortality of third-instar larvae was assessed daily for 7 days after exposure to concentrations of silver nanoparticles, sorghum polymer particles, and hybrid silver-sorghum polymer particles. Both silver nanoparticles and silver-sorghum polymer particles were insecticidal, but sorghum polymer particles alone did not significantly contribute to larval mortality. Concentrations of 100 mg/liter of silver nanoparticles achieved >50% mortality at day 7, and 200 mg/liter treatments achieved >75% larval mortality within 24 h. The antimicrobial properties of silver nanoparticles were also examined, and culturable bacteria were recovered from larval-rearing media at 200 mg/liter but not at 400 mg/liter of silver nanoparticles. These data suggest that C. sonorensis larval mortality is primarily caused by silver nanoparticle toxicity and not by the reduction of bacteria (i.e., a larval food source). This work describes the first use of silver nanoparticles in C. sonorensis and shows the potential insecticide applications of these nanoparticles against this agricultural pest. The grain-polymer particles also successfully carried insecticidal silver nanoparticles, and their utility in loading diverse compounds could be a novel toxin delivery system for biting midges and similar pests., (Published by Oxford University Press on behalf of Entomological Society of America 2024.)

( S )-Methoprene has been widely applied as a powerful biochemical pesticide to control disease vectors and other pestiferous arthropods of economic importance. As a juvenile hormone analogue, many products based on ( S )-methoprene are developed and commercialized in the USA, Europe, and elsewhere. However, the agricultural use of ( S )-methoprene and its analogues remains underexplored. Here, based on an intermediate derivatization strategy and structural modification, a series of enantiopure ( S )-methoprene derivatives were designed for their expected bioactivity against two crop-threatening pests. Six compounds showed more than 2-fold stronger inhibition of emergence against Plutella xylostella than ( S )-methoprene, among which one that was designated as B2 showed even superior activity to the conventional chemical pesticide and biopesticide with IE 50 of 0.02 mg/L. Nine compounds exhibited over 2-fold higher bioactivity against Aphis craccivora growth than ( S )-methoprene. The physicochemical property evaluation and toxicological test showed that the potent ( S )-methoprene derivatives were low toxic to the nontarget organism and the environment. Molecular docking studies further demonstrated that the high bioactivity of B2 may be partially attributed to its great affinity for binding to juvenile hormone receptors of P. xylostella . The current study suggests that B2 is a biochemical pesticide candidate with potency to be developed as a new agrochemical for lepidopteran control.

Although the metabolism of yolk lipids such as docosahexaenoic acid (DHA) is pivotal for embryonic development, the underlying mechanism remains elusive. Here we find that the zebrafish hydroxysteroid (17-β) dehydrogenase 12a (hsd17b12a), which encodes an intestinal epithelial-specific enzyme, is essential for the biosynthesis of long-chain polyunsaturated fatty acids in primitive intestine of larval fish. The deficiency of hsd17b12a leads to severe developmental defects in the primitive intestine and exocrine pancreas. Mechanistically, hsd17b12a deficiency interrupts DHA synthesis from essential fatty acids derived from yolk-deposited triglycerides, and consequently disrupts the intestinal DHA-phosphatidic acid (PA)-phosphatidylglycerol (PG) axis. This ultimately results in developmental defects of digestive organs, primarily driven by ferroptosis. Our findings indicate that the DHA-PA-PG axis in the primitive intestine facilitates the uptake of yolk lipids and promotes the expansion of digestive organs, thereby uncovering a mechanism through which DHA regulates embryonic development., Competing Interests: Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Caste differentiation involves many functional traits that diverge during larval growth and metamorphosis to produce adults irreversibly adapted to reproductive division of labor. Investigating developmental differentiation is important for general biological understanding and has increasingly been explored for social phenotypes that diverge in parallel from similar genotypes. Here, we use Monomorium pharaonis ants to investigate the extent to which canalized worker development can be shifted toward gyne (virgin-queen) phenotypes by juvenile hormone (JH) treatment. We show that excess JH can activate gyne-biased development in workers so that wing-buds, ocelli, antennal and genital imaginal discs, flight muscles, and gyne-like fat bodies and brains emerge after pupation. However, ovary development remained unresponsive to JH treatment, indicating that JH-sensitive germline sequestration happens well before somatic differentiation. Our findings reveal important qualitative restrictions in the extent to which JH treatment can redirect larval development and that these constraints are independent of body size. Our findings corroborate that JH is a key hormone for inducing caste differentiation but show that this process can be asymmetric for higher colony-level germline versus somatic caste differentiation in superorganisms as defined a century ago by Wheeler. We quantified gene expression changes in response to JH treatment throughout development and identified a set of JH-sensitive genes responsible for the emergence of gyne-like somatic traits. Our study suggests that the gonadotropic role of JH in ovary maturation has shifted from the individual level in solitary insects to the colony level in an evolutionary-derived and highly polygynous superorganism like the pharaoh ant., Competing Interests: Competing interests statement:The authors declare no competing interest.

Background: Genetic biocontrol interventions targeting mosquito-borne diseases require the release of male mosquitoes exclusively, as only females consume blood and transmit pathogens. Releasing only males eliminates the risk of increasing mosquito bites and spreading pathogens while enabling effective population control. The aim of this study is to develop robust sex-sorting methods for early larval stages in mosquitoes, enabling scalable male-only releases for genetic biocontrol interventions., Methods: To address the challenge of sex-sorting in the Asian malaria vector Anopheles stephensi, we engineer Sexing Element Produced by Alternative RNA-splicing of a Transgenic Observable Reporter (SEPARATOR). This dominant fluorescent-based method, previously proven effective in Aedes aegypti, exploits sex-specific alternative splicing of a reporter to ensure exclusive male-specific expression early in development. The sex-specific alternative RNA splicing of the doublesex gene was selected as a target for engineering SEPARATOR due to its evolutionary conservation in insects. To expand SEPARATOR's applicability for genetic sexing, we assessed the cross-species sex-specific RNA splicing activity of the An. gambiae doublesex (AngDsx) splicing module in An. stephensi. Male-specific enhanced green fluorescent protein (EGFP) expression was verified throughout the mosquito life cycle using a fluorescent stereomicroscope., Results: Our results confirm that SEPARATOR regulates male-specific EGFP expression in An. stephensi and enables reliable positive male selection from the first instar larval stages. Molecular analysis demonstrates that male-specific EGFP expression is dependent on doublesex sex-specific splicing events. Additionally, the splicing module from An. gambiae operates effectively in An. stephensi, demonstrating evolutionary conservation in sex-specific splicing events between these species., Conclusions: SEPARATOR's independence from sex-chromosome linkage provides resistance to breakage that could be mediated by meiotic recombination and chromosomal rearrangements, making it highly suitable for mass male releases. By enabling precise male selection from the first instar larval stages, SEPARATOR represents a significant advancement that will aid in the genetic biocontrol for Anopheles mosquitoes., Competing Interests: Declarations Ethical approval and consent to participate All animals were handled in accordance with the Guide for the Care and Use of Laboratory Animals as recommended by the National Institutes of Health and approved by the UCSD Institutional Animal Care and Use Committee (IACUC, Animal Use Protocol #S17187) and UCSD Biological Use Authorization (BUA #R2401). No human research was conducted so we did not need consent to participate in this study. Consent for publication Not applicable. Competing interests O.S.A. and S.W. have filed a patent on this technology. O.S.A is a founder of Agragene, Inc. and Synvect, Inc. with equity interest. The terms of this arrangement have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies. All other authors declare no competing interests., (© 2024. The Author(s).)

Deoxygenation is a growing threat to marine ecosystems, with an increase in the frequency, extent and intensity of hypoxia events in recent decades. These phenomena will pose various challenges to marine species, as it affects their survival, growth, body condition, metabolism and ability to handle other environmental stressors, such as temperature. Early life stages are particularly vulnerable to these changes. Thus, it is crucial to understand how these initial phases will respond to hypoxia to predict the impacts on marine populations and ecosystems. In this work, we aimed to evaluate the effect of oxygen (O 2 ) availability on fitness related traits (mortality, growth and body condition), metabolism (Routine metabolic rates [RMR]) and thermal tolerance (CTmax), in early stages of Atherina presbyter, exposed for two weeks, to two O 2 levels: normoxia (6.5-7.2 mg L -1 ) and hypoxia (2-2.5 mg L -1 ), through an experiment setup. Our findings showed that while low oxygen levels did not negatively impact mortality, total length, weight, or body condition (Fulton K), the larvae undergo metabolic depression when exposed to hypoxia, as an energy conservation mechanism. Furthermore, CTmax suffered a significant reduction in low O 2 availability, due to the inability of the circulatory and respiratory systems to fulfill energy demands. These outcomes suggest that although early life stages of Atherina presbyter can survive under low oxygen environments, they are less capable of dealing with sudden increases in temperature when oxygen is scarce., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Larval source management (LSM) effectively reduces mosquito populations at their breeding sites, addressing issues like insecticide resistance that limit the effectiveness of primary interventions such as insecticide-treated nets (ITNs). Although traditionally used in urban and dry areas, recent research suggests it might also be effective in rural settings in eastern and southern Africa, where Anopheles funestus thrives in permanent water bodies that sustain year-round transmission. Targeting these habitats could enhance LSM, but it requires understanding local community practices, as mosquito breeding sites often overlap with community water resources. This study examined how communities use aquatic habitats and how these practices may impact LSM strategies, with a focus on habitats used by An. funestus., Methods: This study was conducted in three villages in the Ulanga and Malinyi districts of southeastern Tanzania using a mixed-methods approach. Quantitative data were collected through cross-sectional surveillance, and qualitative data through unstructured interviews, focus group discussions, and field observations. Data analysis integrated both quantitative and qualitative findings to develop a comprehensive understanding of community perspectives., Results: A survey of 931 aquatic habitats found mosquito larvae in 73% of them, with late instar An. funestus present in 23%. River streams made up 41% of the habitats, while ground pools accounted for 4%. Most habitats (90%) were used by communities, including 95% of those with An. funestus larvae, for activities such as domestic chores, agriculture, livestock rearing, brickmaking, and fishing. Focus group discussions revealed a willingness to adopt LSM, with a preference for larviciding and habitat modification over habitat removal, as the water sources were vital for daily use. Community concerns centered on the safety of larvicides for humans and animals, the environmental impact, and the need for better awareness of how LSM affects health and livelihoods., Conclusion: This study highlights community perspectives on LSM, focusing on the dual function of aquatic habitats as mosquito breeding sites and essential community water sources. This dual role presents both challenges and opportunities, suggesting that LSM strategies must balance public health needs with socio-economic realities. There was a clear preference for larviciding and habitat modification over removal, with a strong emphasis on health and environmental safety. The study emphasizes the importance of educating communities on the safety and effectiveness of LSM, and tailoring LSM strategies to fit the needs and preferences of local communities., (© 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).)

Introduction: Pintomyia (Pifanomyia) longiflocosa is an endemic species from Colombia, found between the central and eastern Andes, and reported as one of the primary vectors of cutaneous leishmaniasis in coffee-growing zones of the country. This species is classified in the Townsendi series and can only be identified by the morphology of the male adults., Objective: To determine the potential use of the fourth larval stage of the vector Pi. longiflocosa in morphological taxonomy based on the description of its chaetotaxy., Materials and Methods: Pintomyia longiflocosa adults were captured in Campoalegre, Huila, and reared in the Entomology Laboratory at the Colombian Instituto Nacional de Salud. To identify the setae found in each corporal segment, 15 fourth-instar larvae were mounted on microscope slides using Canadian balm after being cleared with 10 % potassium hydroxide and saturated phenol. Additionally, five specimens were prepared for their observation by scanning electron microscopy., Results: Based on the description of Pi. longiflocosa, we established that all species of the subgenus Pifanomyia so far described have the same antennal morphology and clavate setae along their body. However, various setae present in Pi. longiflocosa are absent in Pi. youngi, suggesting differences among the larvae of the Townsendi series., Conclusions: These results support the potential importance of morphological characters from the fourth larval instar, such as antennal morphology and chaetotaxy, specifically in closely related species that are cryptic in their adult stages.

The main objective of this study was to investigate the effects of commercial probiotic Bacillus sp. supplementation on seabream Sparus aurata larviculture under culture conditions. In this context, Bacillus was supplemented via rotifer feeding and water and its effects on pancreatic and intestinal enzyme activities as well as aquaculture parameters were evaluated during early life development. In the experimental group, as probiotic three Bacillus sp. spores were introduced via rotifer and larval culture tanks, while the larvae in control group did not feed any probiotic supplementation. At the end of the experiment on 40 days after hatching, the probiotic-supplemented group exhibited better growth performance and there were statistically differences in between groups of probiotic-treated and control regarding growth parameters (P < 0.01), despite insignificant survival rate (P > 0.05). In terms of enzymatic expressions, S. aurata larvae receiving probiotic supplementation through rotifers demonstrated noteworthy (P < 0.05) enhancements in specific activities of pancreatic and intestinal enzymes, except for amylase (P > 0.05), when compared to the control group. It is concluded that the administration of Bacillus sp. as probiotic bacteria through rotifer supplementation and water intake demonstrates significant positive impacts on both growth parameters and specific activities of main pancreatic and intestinal enzymes of seabream larvae., (© The Author(s) 2024. Published by Oxford University Press on behalf of Applied Microbiology International.)

In recent years, isoxazole insecticides or parasiticides targeting the γ-aminobutyric acid receptor, such as fluralaner or fluxametamide, featured a novel chemical structure and exhibited potent insecticidal activity with no-cross resistance. Thus, many research institutes have tried to modify the structures of these agents to find a new insecticide. Previously, the majority of researchers stuck to the "phenyl-isoxazole-phenyl" structure, making modifications only to other components. In this study, the "phenyl-isoxazole-phenyl" ternary motif was modified for the first time based on bioisosterism theory. A series of new derivatives carrying pyrazoline and 1,3,4-oxadiazole moieties were designed and synthesized to investigate their insecticidal activities against the diamondback moth ( Plutella xylostella ) and fall armyworm ( Spodoptera frugiperda ). Preliminary bioassay data showed that some of the target compounds exhibited good insecticidal activities against P. xylostella and S. frugiperda . Especially, compound A21 showed insecticidal activity against P. xylostella (LC 50 = 1.2 μg/mL) better than commercial insecticide ethiprole (LC 50 = 2.9 μg/mL) but worse than parasiticide fluralaner (LC 50 = 0.5 μg/mL). Similarly, compound A21 exhibited insecticidal activity to S. frugiperda (LC 50 = 13.2 μg/mL) better than commercial insecticide fipronil (LC 50 = 78.8 μg/mL) but worse than fluralaner (LC 50 = 0.7 μg/mL). Compound A21 could serve as a potential lead compound to control P. xylostella and S. frugiperda . The three-dimensional quantitative structure-activity relationship model revealed that the further introduction of an electron-donating group in the 2- or 3-site may increase the insecticidal activity of A21 . Molecular dynamics simulations showed that the hydrogen bond of A21 and receptor was important for the binding receptor. This study has identified a new substructure called "phenyl-pyrroline-oxadiazole" instead of the previously known "phenyl-isoxazole-phenyl" substructure, offering a useful guide for the design of novel insecticide molecules.

In French Polynesia, the pearl farming industry relies entirely on collecting natural spat using a shade-mesh collector, which is reported to contribute to both plastic pollution and the release of toxic chemicals. With the aim of identifying more environment-friendly collectors, this study investigates the chemical toxicity of shade-mesh (SM) and alternative materials, including reusable plates (P), a newly developed biomaterial (BioM) and Coconut coir geotextile (Coco), on the embryo-larval development of Pinctada margaritifera. Embryos were exposed during 48 h to four concentrations (0, 0.1, 10 and 100 g L -1 ) of leachates produced from materials. Chemical screening of raw materials and leachates was performed to assess potential relationships with the toxicity observed on D-larvae development. Compared to the other tested materials, results demonstrated lower levels of chemical pollutants in BioM and no toxic effects of its leachates at 10 g L -1 . No toxicity was observed at the lowest tested concentration (0.1 g L -1 ). These findings offer valuable insights for promoting safer spat collector alternatives such as BioM and contribute to the sustainable development of pearl farming., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

MicroRNAs (miRNAs) are post-transcriptional, non-coding regulatory RNAs that function coordinately with transcription factors (TFs) in gene regulatory networks. TFs and their targets are often co-regulated by miRNAs, forming composite feedforward circuits (cFFCs) with varying degrees of redundancy, primarily mediated by miRNAs. However, the maintenance of miRNA-mediated regulatory redundancy and its impact on gene expression evolution remain elusive. By integrating ChIP-seq data from ENCODE and miRNA targeting from TargetScanFly, we quantified miRNA-mediated cFFC redundancy in Drosophila melanogaster embryos and larvae, revealing more than three quarters of miRNA targets are involved in redundant cFFCs. Higher cFFC redundancy, where more miRNAs target the same gene within a cFFC, is correlated with stronger purifying selection, reduced expression divergence between species, and increased expression stability under heat shock stress. Redundant cFFCs primarily regulate older or broadly expressed young genes. These findings highlight the role of miRNA-mediated cFFC redundancy in enhancing gene expression robustness through natural selection., (© 2024. The Author(s).)

Background: The gut microbiota can facilitate host growth under nutrient-constrained conditions. However, whether this effect is limited to certain bacterial species remains largely unclear, and the relevant underlying mechanisms remain to be thoroughly investigated., Results: We found that the microbiota was required for Bactrocera dorsalis larval growth under poor dietary conditions. Monoassociation experiments revealed that Enterobacteriaceae and some Lactobacilli promoted larval growth. Among the 27 bacterial strains tested, 14 significantly promoted larval development, and the Enterobacteriaceae cloacae isolate exhibited the most obvious promoting effect. A bacterial genome-wide association study (GWAS) revealed that the vitamin B6 synthesis pathway was critical for the promotion of E. cloacae growth. Deletion of pdxA, which is responsible for vitamin B6 biosynthesis, deprived the mutant strains of larval growth-promoting function, indicating that the 4-hydroxythreonine-4-phosphate dehydrogenase(pdxA) gene was crucial for promoting larval growth in E. cloacae. Importantly, supplementation of a poor diet with vitamin B6 successfully rescued the axenic larval growth phenotype of B. dorsalis., Conclusion: Our results suggest that gut microbes promote insect larval growth by providing vitamin B6 under nutrient scarcity conditions in B. dorsalis. Video Abstract., (© 2024. The Author(s).)

Aims: A wide range of vector control programmes rely on the efficient production and release of male mosquito. Asaia bacteria are described as potential symbionts of several mosquito species but their relationship with Aedes aegypti has never been rigorously tested. Here, we aimed to quantify the benefits of three Asaia species on host development in Ae. aegypti, and the ability of these bacteria to form a stable symbiotic association with growing larvae., Methods and Results: In order to disentangle direct and indirect effects of Asaia inoculation on host development, experiments used insects with an intact microbiome and those reared in near-aseptic conditions, while we characterized bacterial communities and Asaia densities with culture dependent and independent methods (16S rRNA amplicon sequencing). Neonate larvae were inoculated with Asaia spp. for 24 h, or left as uninoculated controls, all were reared on sterile food. Aseptic larvae were produced by surface sterilization of eggs. Although all Asaia were transient members of the gut community, two species accelerated larval development relative to controls. The two mutualistic species had lasting impacts on the larval microbiome, largely by altering the relative abundance of dominant bacteria, namely Klebsiella and Pseudomonas. Axenic larvae were dominated by Asaia when inoculated with this species but showed slower development than conventionally reared insects, indicating that Asaia alone could not restore normal development., Conclusions: Our results reveal Asaia as a poor mutualist for Ae. aegypti, but with a species-specific positive effect on improving host performance mediated by interactions with other bacteria., (© The Author(s) 2024. Published by Oxford University Press on behalf of Applied Microbiology International.)

The operation of the retina, like other brain circuits, is under modulatory control. One coordinator of changes in retinal function is dopamine, a neuromodulator released in a light-dependent way to adjust vision on a diurnal cycle. Here, we demonstrate that substance P is a similarly powerful retinal modulator that interacts with the dopamine system. By imaging glutamatergic synaptic transmission in larval zebrafish, we find that substance P decreases the contrast sensitivity of ON and OFF visual channels up to 8-fold, with suppression of visual signals being strongest through the "transient" pathway responding to higher frequencies. These actions are exerted in the morning, in large part by suppressing the amplification of visual signals by dopamine, but substance P is almost completely inactive in the afternoon. Modulation of retinal gain is accompanied by changes in patterns of vesicle release at the synapses of bipolar cells: increased gain shifts coding of stimulus strength from the rate of release events to their amplitude generated by a process of multivesicular release (MVR). Together, these actions of substance P reduce the flow of visual information, measured in bits, ∼3-fold. Thus, whereas dopamine "pushes" the retina to transmit information at higher rates in the afternoon, substance P acts in antiphase to suppress dopamine signaling and "pull down" information transmission in the morning., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Honeybee larvae and pupae form the cornerstone of colony survival, development, and reproduction. Azoxystrobin is an effective strobilurin fungicide that is applied during the flowering stage for controlling plant pathogens. The contaminated nectar and pollen resulting from its application are collected by forager bees and impact the health of honeybee larvae and pupae. The current study evaluated the survival, development, and physiological effects of azoxystrobin exposure on the larvae and pupae of Apis mellifera worker bees. The field-recommended concentrations of azoxystrobin were found to suppress the survival indices and lifespan in the larval as well as pupal stages; moreover, the rates of the survival and pupation of larvae as well as the body weights of the pupae and newly-emerged adult bees were significantly reduced upon long-term exposure to azoxystrobin. In addition, azoxystrobin ingestion induced changes in the expression of genes critical for the development, immunity, and nutrient metabolism of larvae and pupae, although the expression profile of these genes differed between the larval and pupal stages. Results indicated the chronic toxicity of azoxystrobin on the growth and development of honeybee larvae and pupae, which would affect their sensitivity to pathogens and other external stresses during the development stage and the study will provide vital information regarding the pollination safety and rational use of pesticides.

Dietary change can be a strong evolutionary force and lead to rapid adaptation in organisms. High-fat and high-sugar diets can challenge key metabolic pathways, negatively affecting other life history traits and inducing pathologies such as obesity and diabetes. In this study, we use experimental evolution to investigate the plastic and evolutionary responses to nutritionally unbalanced diets. We reared replicated lines of larvae of the housefly Musca domestica on a fat-enriched (FAT), a sugar-enriched (SUG), and a control (CTRL) diet for thirteen generations. We measured development time in each generation and larval growth and fat accumulation in generations 1, 7, and 13. Subsequently, all lines were reared for one generation on the control diet to detect any plastic and evolutionary changes. In the first generation, time to pupation decreased on a fat-rich diet and increased on a sugar-rich diet. The fat-rich diet increased fat accumulation and, to a lesser extent, the dry weight of the larvae. Multigenerational exposure to unbalanced diets caused compensatory changes in development time, dry weight, and absolute and relative fat content, although pattern and timing depended on diet and trait. When put back on a control diet, many of the changes induced by the unbalanced diets disappeared, indicating that the diet has large plastic effects. Nevertheless, fat-evolved lines still grew significantly larger than the sugar-evolved lines, and sugar-evolved lines had consistently lower fat content. This can be an effect of parental diet or an evolutionary change in nutrient metabolism as a consequence of multigenerational exposure to unbalanced diets., (© The Author(s) 2024. Published by Oxford University Press on behalf of the European Society of Evolutionary Biology.)

The caloric content and macronutrient ratio of diet consumed is a major source of phenotypic variation in most animal populations. While these nutritional effects have been well-documented for a variety of life-history and morphological traits, the effects of nutrition on male genitals are poorly understood but genitals are thought to be more canalised than general morphology and hence less susceptible to variation in nutrition. Even less is known about the effects of nutrition on female genital form, which to our knowledge, have never been investigated. Here we tested for effects of juvenile dietary macronutrients (protein and carbohydrate) on larval survival, adult morphology, including genital size and shape in male and female flour beetles (Tribolium castaneum). We found there was nutritionally induced plasticity in larval survival and morphology, although the latter effect was variable, with body size being most responsive to dietary macronutrients and genital size and shape being least responsive. Functionally equivalent morphological traits in the sexes responded similarly to nutrition. Previously, we showed that the genitalia of male and female T. castaneum are subject to strong stabilising sexual selection, and our current findings suggest that developmental mechanisms reduce the nutritional sensitivity of male and female genitals, possibly to ensure matching during mating., (© The Author(s) 2024. Published by Oxford University Press on behalf of the European Society of Evolutionary Biology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The predaceous fly Neoditomyia farri Coher is currently regarded as having an unknown population distribution in Jamaica. The larvae are known for their production of sticky "fishing lines" that are covered in adhesive droplets for prey capture and hang down from cave walls and ceilings. Published research beyond early observational records of the species is limited to one site-Dromilly Cave in the parish of Trelawny. The Jamaican Caves Organisation has observed larvae of the species at 26 caves visited over the last 2 decades and compiled a database with details of approximate numbers, internal and external environmental conditions, and accurate positions. Based on these records, we present evidence that the species is not cave obligate and describe factors that may suggest potential additional sites where the species can be found. Taken together, our observations paint a picture of a species which is ubiquitous and not under discernible threat, making it an excellent candidate for further research on its unique life history and behavior, which remain poorly understood., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America.)

Climate change is rapidly warming thermal environments, an important abiotic stimulus governing interactions between microbial symbionts and their hosts. Increasing evidence suggests that solitary bees rely on pollen provision microbes for successful development. However, the effects of heat stress on provision microbiota and the resulting consequences for larval health and development remain to be examined. We performed an in vitro study to investigate the effects of the thermal environment on provision microbiome composition and measured fitness outcomes for Osmia lignaria larvae. While pollen sterilisation removed bacteria from microbe-rich provisions, larval survivorship did not significantly differ between bees reared on microbe-rich (unmanipulated) diets and provisions treated with ethylene oxide (EO) gas. In contrast to previous research in solitary bees, larvae reared on EO-treated provisions weighed more and had higher total fat content, with temperature moderating the degree of difference. As anticipated, we observed a negative relationship between the duration of larval development and temperature. Our results indicated that an intact provision microbiota may not always improve bee fitness and that bee-microbe interactions during larval development may contribute to the size-shrinking effect observed for cavity-nesting bees under warming conditions., (© 2024 John Wiley & Sons Ltd.)

Early life stages of fish are insufficiently studied in most regions around the globe, and consequently, there is an important gap in knowledge on the life cycle of most teleosts. In the Canary Islands (Eastern Central Atlantic), most studies on ichthyoplankton communities are based on traditional net tows in open waters that provide information about early larval stages of pelagic species in the area, while nearshore benthic fish remain highly understudied. In this study, light traps were employed for the first time to assess post-larval stages of neritic fish in this archipelago. A two-year survey was carried out to collect nearshore fish larvae every 6 months at 11 localities from El Hierro, Tenerife and Lanzarote. A total of 3940 fish larvae classified into 13 orders, 28 families and 44 species were collected. These results provide a wide description of the composition of the inshore ichthyoplankton off the Canary Islands, across islands and seasons. Main environmental factors (SST, sea floor orography, oceanographic phenomena …) influencing the population dynamics of this community are discussed. Additionally, the ichthyoplankton assemblages were assessed from the intra-annual perspective, analyzing the species composition and abundances by months and seasons, and providing new insights into reproduction cycles of many common benthic fishes at shallow marine ecosystems of the islands. This study is important to further understand the life cycles of some of the most common fish species in the Canary Islands, unraveling main environmental factors that affect the success of their offspring that sustain populations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The Arabian goby, Cryptocentroides arabicus (Gobiidae: Gobiinae), is a significant species in marine and brackish water ecosystems. However, limited information is available about the reproductive biology and the early life history of this species. This study aimed to understand the embryonic and pre-larval development of C. arabicus under controlled laboratory conditions using light microscopy. Ten fish pairs were placed in five 70-L glass aquaria, and environmental parameters were carefully monitored. After 4 months, spawning began, with each event yielding between 700 and 1500 eggs, totaling 10 spawning events. The elliptical-shaped eggs, measuring 3.9 ± 0.1 mm in length and 1.6 mm in maximum width, hatched after 7 days and 4 h at a water temperature of 28°C. This process resulted in the identification of eight distinct larval developmental stages. Parental care by both the male and female was observed during the incubation period. Documenting the morphological changes during incubation contributes to a better understanding of the reproductive behaviour and early life history of C. arabicus. This is essential for the conservation management of marine and brackish water environments., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Recruitment success depends on external forcing mechanisms such as ocean currents that affect the transport of eggs and larvae to favorable habitats. In this study, we investigated the role of larval transport in the recruitment of Mullus barbatus in the Central Mediterranean Sea by modeling the recruits' abundance as a function of both spawning stock size and dispersal rates of the species' early life stages. Our analysis involved twenty years of data on recruits and spawners abundance obtained from scientific trawl surveys, and data on larval dispersal rates derived from a combination of actualized published sources and original data. By calculating the estimates of retention, import and uniformity of the contribution of the spawning areas distributed among different Geographical Sub Areas (GSAs) in the Sicilian nurseries, we assessed their contribution to recruitment using modified Ricker stock size-recruits models. In particular, our results show that a high uniform contribution from spawning areas within GSA16, mainly related to the oceanographic patterns promoting larval retention, together with spawners abundance, significantly reduced the variability of red mullet recruitment. We further highlighted that when switching from a higher to a lower level of evenness of contribution to the recruit population from different spawning areas in the GSA16, the expected spawning stock abundance per recruit for a given fishing pattern can suffer a rapid short-term decline, which is likely to have negative consequences for stock assessment and management decisions. Our results suggest that larval transport plays a crucial role in explaining the interannual variability of recruitment, thereby contributing to a better understanding of stock size variation. Additionally, our study enhances the understanding of the spatial dynamics involved in the recruitment of this species, which is of increasing interest within fisheries management frameworks., Competing Interests: Declaration of competing interest We confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome.All authors (FQ, FF, FG and GG) contributed to the study conception and design. Material preparations were performed by FQ, FG the formal analysis were performed by FQ and FG. FQ: Writing – original draft. All authors read and approved the final manuscript., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The neuropeptide prothoracicotropic hormone (PTTH) plays a key role in regulating ecdysone synthesis and promoting insect metamorphosis. Pyriproxyfen is a juvenile hormone analogue. We previously reported that pyriproxyfen disrupts ecdysone secretion and inhibits larval-pupal metamorphosis in silkworms. However, the specific molecular mechanisms by which pyriproxyfen interferes with ecdysone signaling remain to be elucidated. Herein, the RNA-seq analysis on the ecdysone-secretion organ prothoracic gland (PG) was conducted following pyriproxyfen exposure. A total of 3774 differentially expressed genes (DEGs) were identified, with 1667 up-regulated and 2107 down-regulated. KEGG analysis showed that DEGs were enriched in the MAPK signaling pathway, a conserved pathway activated by PTTH binding to Torso, which regulates the ecdysone synthesis. qRT-PCR results indicated a significant up-regulation in PTTH transcription level, while the transcription levels of torso and downstream MAPK pathway genes, Ras2, Raf and ERK, were down-regulated 24 h post-pyriproxyfen treatment. Consistent with these transcriptional changes, PTTH titers in the brain also increased following pyriproxyfen treatment. These results suggest that pyriproxyfen induces abnormal metamorphosis in silkworms by impairing PTTH-Torso signaling. This study enhances our understanding of the molecular mechanisms of pyriproxyfen-induced larval-pupal abnormal metamorphosis in silkworms, and also provides insights for developing detoxification strategies for juvenile hormone analog pesticides to non-target organisms., Competing Interests: Declaration of competing interest The authors that there are no conflicts of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The glutamate-gated chloride channels (GluCls) are widely existed in the neural and nonneural tissues of invertebrate. In addition to play important roles in signal transduction, the GluCls also showed multiple physiological functions in insects such as participate in the juvenile hormone synthesis. In the present study, the potential roles of TcGluCl in growth and development of the red flour beetle Tribolium castaneum were explored. Knockdown of TcGluCl showed no effects on the survivability, weight growth, final pupation rate, eclosion and fecundity of T. castaneum, whereas resulted in the significant premature pupation of larvae. Inhibition of TcGluCl expression significantly changed the levels of juvenile hormone and ecdysone as well as the expressions of hormone biosynthetic genes. The increased ecdysone level and decreased juvenile hormone level were observed at the late stage of dsGluCl-treated larvae. Knockdown of TcGluCl significantly reduced the expressions of TcSTIM1 and TcOrai1, which were the primary proteins in store-operated calcium entry (SOCE) mediated Ca 2+ influx mechanism. Whilst the L-glutamic acid treatment led to the increased TcOrai1 expression in T. castaneum. These findings suggested that knockdown of TcGluCl increased the ecdysone level and contributed to the premature pupation of larvae, which might be due to the reduced Ca 2+ influx caused by the decreased expressions of TcSTIM1 and TcOrai1. These studies provide novel insights on the function of GluCls in insects., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Spodoptera frugiperda is an economic agricultural pest that has invaded many countries around the world and caused huge losses in grain production. Camptothecin (CPT) is one of the botanical compounds with insecticidal activity and has the potential for pest control. However, the effects of CPT on development and metabolism of S. frugiperda remain unknown. In this study, we have investigated the adverse effects of 1.0 and 5.0 mg/kg CPT exposures on the growth and development of S. frugiperda. Our results found that 1.0 and 5.0 mg/kg CPT treatments altered the parameters of the life cycle, including inducing larval mortality, altering the weight of larvae, pupae, and adults, the larval duration, and decreasing the pupation rate and emergence rate. In addition, comparative metabolomics analysis was performed in the larval midgut of S. frugiperda to explore the toxicity mechanism of CPT. A total of 261 and 348 differential metabolites were identified in the groups with 1.0 and 5.0 mg/kg CPT treatments, respectively. Further analysis found that pantothenate and CoA biosynthesis, sulfur relay system, selenocompound metabolism, and fatty acid biosynthesis pathways were significantly altered by 5.0 mg/kg CPT exposure. Our results provided new insight into the toxicological mechanisms of CPT against S. frugiperda and laid the foundation for the field application of CPT in pest control., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Black soldier fly larvae (BSFL; Hermetia illucens) hold promise for converting biowaste into proteins and lipids for feed. Dietary starch is efficiently digested by the larvae and influences larval performance, but the mechanisms of starch digestion remain poorly understood. This study investigated changes in individual weight and amylase activity in BSFL after 4, 7 and 11 days of feeding for five substrates varying in starch content and type: chicken feed (CF), corn gluten feed (CGF), wheat bran (WB), wheat distillers grain (WDG) and discarded potatoes (DP). Substrate amylase activities were also measured with and without larvae (feeding and fermenting trays, respectively) over time in order to explore external digestion. Feed conversion ratio (FCR) and estimated digestibility (ED) of DM and starch were assessed at the end of the experiment. The ranking for best FCR was CF, WB, CGF, WDG and DP. In feeding trays, ED of DM was 69.8 ± 1.8, 59.5 ± 2.9, 58.6 ± 0.7, 45.4 ± 0.6 and 19.5 ± 0.8% in CF, DP, WB, CGF and WDG, respectively. Estimated digestibility of starch reached 100% with WB and CGF, followed by CF (88.2 ± 2.3%), DP (85.2 ± 1.2%) and WDG (43.1 ± 1.0%). Larval amylase activity increased with growth for all substrates and dropped when approaching pupation. No relationship was found between larval amylase activity and substrate starch or other nutrient content, but a negative correlation was reported with the reducing sugar content of the larvae, suggesting glucose repression of amylase production. Amylase activity decreased with time in all feeding and fermenting substrates except WDG and DP. In vitro degradation assays indicated that BSFL amylase was nine times more efficient on raw corn or wheat starch than on raw potato starch, highlighting that starch structure is a major driver of digestibility. Western blot analysis revealed the presence of BSFL amylase in the feeding substrate, hinting at external digestion. Larval amylase was purified to identify its optimal pH (5.0-6.5) and temperature (70 °C). These results highlight that starch content is not a major driver of amylase activity in BSFL and suggest that other non-investigated factors could have had a crucial impact on the activity of larval digestive enzymes, such as microbial community of the substrate and presence of amylase inhibitors. This study also provides insights into the evolution of BSFL digestive activity during their development and the occurrence of external digestion., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Parasitoids often exhibit high flexibility in their development depending on stages of their host at the parasitism, yet little is known about the mechanism underlying such flexibility. In the study, we evaluated the larval development time of the parasitoid Exorista sorbillans (Diptera: Tachinidae) on the lepidopteran model insect Bombyx mori (Lepidoptera: Bombycidae). We found that the development duration of E. sorbillans larvae parasitizing on the late-developmental silkworms was significantly shorter than that of the larvae parasitizing on the early-developmental hosts. Intriguingly, the 2nd-3rd instar molting of parasitoid always occurred when the ecdysteroid titers in the host were increased to higher levels. Furthermore, inhibiting the release of ecdysteroids to parasitic abdomen by thorax-abdomen ligation of the host only repressed the 2nd-instar growth and molting of E. sorbillans larvae, but had no effect on their pupation. Meanwhile, the ecdysone synthesis and 20-hydroxyecdysone (20 E) signaling in larval parasitoids were impeded after ligation treatment. Moreover, exogenous 20 E application could largely rescue the defect in 2nd instar growth and molting through stimulating ecdysone synthesis and signaling in E. sorbillans. Our results indicate that the parasitoid requires the host ecdysteroids to stimulate 20 E signaling and the subsequent 2nd-instar growth and molting. These findings will improve our understanding of the host utilization strategies of parasitoids, and contribute to the development of in vitro rearing procedures of tachinid parasitoids for biological control., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Climate change facilitates the rapid invasion of agricultural pests, threatening global food security. The fall armyworm Spodoptera frugiperda is a highly polyphagous migratory pest tolerant to high temperatures, allowing its proliferation in harsh thermal environments. We aimed to demonstrate mechanisms of its high-temperature tolerance, particularly transcriptional and metabolic regulation, which are poorly understood. To achieve the aim, we examined the impact and mechanism of heat events on S. frugiperda by using multiple approaches: ecological measurements, transcriptomics, metabolomics, RNAi, and CRISPR/Cas9 technology. We observed that several physiological indices (larval survival rate, larval period, pupation rate, pupal weight, eclosion rate, and average fecundity) decreased as the temperature increased, with the 32 °C treatment displaying a significant difference from the control group at 26 °C. Significantly upregulated expression of genes encoding endochitinase and chitin deacetylase was observed in the chitin-binding, extracellular region, and carbohydrate metabolic process GO terms of hemolymph, fat body, and brain, exhibiting a tissue-specific pattern. Significantly enriched pathways (e.g., cutin, suberin, and wax biosynthesis; oxidative phosphorylation and cofactor biosynthesis; diverse amino acid biosynthesis and degradation; carbon metabolism; and energy metabolism), all of which are essential for S. frugiperda larvae to tolerate temperature, were found in metabolites that were expressed differently. Successful RNA interference targeting of the three chitin-related genes reduced gene expression levels and larval survival rate. Knockout of the endochitinase gene by using the CRISPR/Cas9 system significantly reduced the relative gene expression and increased sensitivity to high-temperature exposure. On the basis of our findings, theoretical foundations for understanding the high-temperature tolerance of S. frugiperda populations and latent genetic control strategies were established., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Drosophila NF-κB transcription factor Dif has been well known for its function in innate immunity, and recent study also reveals its role in neuronal cells. However, the underlying mechanisms of Dif in the brain remain elusive. In this study, we aim to investigate the function of Dif in Drosophila brain development and how Dif regulates structure and plasticity of the brain to affect aging and behaviors. Based on the analysis of differentially expressed genes, we identified key genes associated with cell division, development and aging in the brain of Dif 1 loss of function mutant. In Dif 1 larvae, we found that the metamorphosis and brain development were delayed, and cell division was decreased. In Dif 1 adults, the number of neuron cells was reduced in the brain, the lifespan and locomotor activity were decreased, protein markers associated with aging-related neurodegenerative diseases in the brain were altered in abundance or activity. Our results indicated that Dif plays a crucial role in brain plasticity and neurogenesis, dysfunction of Dif delays larval brain development and impacts proliferation of neuronal cells, resulting in aging adult brain by regulating expression of key genes in multiple signaling pathways involved in cell division, neurogenesis and aging., Competing Interests: Declaration of competing interest The authors have declared that no competing interests exist., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Salivary proteins in the oral secretion (OS) of chewing insects play a crucial role in insect-plant interactions during feeding. The rice leaf folder Cnaphalocrocis medinalis, a notorious pest in global rice production, triggers defense responses during feeding, but little is known about its salivary proteins. In this study, we confirmed that C. medinalis releases OS during feeding. By employing transcriptomic analysis and liquid chromatography-tandem mass spectroscopy (LC-MS/MS), we examined the salivary proteins from labial salivary glands and OS from C. medinalis. A total of 14,397 genes were expressed at the RNA level and 229 salivary proteins were identified. Comparative analysis with other 25 arthropod species revealed that 43 proteins were unique to C. medinalis. Expression pattern analysis revealed that most of the selected genes were highly expressed in the gut and the larval stages (4th-5th instar). These findings provide a comprehensive resource for future functional studies of salivary proteins, offering new insights into the molecular mechanisms by which C. medinalis modulates plant defenses and potential applications in pest management., Competing Interests: Declaration of competing interest All authors have declared that they have no potential competing interests both in personally and financially., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The use of Black Soldier Fly (BSF) larvae is emerging as a promising alternative for biowaste (i.e. food waste) treatment, generating larval biomass and process residues, suitable for use as animal feed and fertilizer, respectively. In line with an increasing use of starch-based bioplastics in food packaging, the presence of these biopolymers and associated biodegradable microplastics (BMPs) in food waste is expected to rise. Knowledge of the generation of BMPs and their fate in the BSF treatment process is scarce, or indeed, completely lacking in the case of small-sized BMPs (<50 μm). The present study aims to investigate the generation and potential accumulation of BMPs in BSF larvae process. Food waste mixed with starch-based bioplastic films was fed to larvae and BMPs of two particle sizes (inferior to and exceeding 10 μm in diameter) were monitored over time in rearing substrate and larval biomass. BMPs concentrations in substrate were compared with larvae-free control tests. The presence of larvae favoured the generation of BMPs. Concentrations of smaller-sized BMPs (<10 μm) increased by approximately 172% in the final substrate, and accumulated in the larval biomass with a peak exceeding the initial larval concentration by over 1000% just before prepupation, which is the typical stage they are collected when used as animal feed. These results indicate a potential risk of soil contamination by BMPs when final substrate is used as fertilizer and a risk of biomagnification phenomena when larvae are used as animal feed., Competing Interests: Declaration of competing interest none., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In a global change scenario, ocean warming and pathogen infection can occur simultaneously in coastal areas, threatening marine species. Data are shown on the impact of temperature on early larvae of the Mediterranean mussel Mytilus galloprovincialis. Increasing temperatures (18-20-22 °C) altered larval phenotypes at 48 hpf and affected gene expression from eggs to 24 and 48 hpf, with shell biogenesis related genes among the most affected. The effects of temperature on larval susceptibility to infection were evaluated using Vibrio coralliilyticus, a coral pathogen increasingly associated with bivalve mortalities, whose ecology is affected by global warming. Malformations and mortalities at 48 hpf were observed at higher temperature and vibrio concentrations, with interactive effects. In non-lethal conditions, interactions on gene expression at 24 and 48 hpf were also detected. Although temperature is the main environmental driver affecting M. galloprovincialis early larvae, warming may increase the susceptibility to vibrio infection, with consequences on mussel populations., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Polyphenisms occur when phenotypic plasticity produces morphologically distinct phenotypes from the same genotype. Plasticity is maintained through fitness trade-offs which are conferred to different phenotypes under specific environmental contexts. Predicting the impacts of contemporary climate change on phenotypic plasticity is critical for climate-sensitive animals like amphibians, but elucidating the selective pressures maintaining polyphenisms requires a framework to control for all mechanistic drivers of plasticity. Using a 32-year dataset documenting the larval and adult histories of 717 Arizona tiger salamanders (Ambystoma mavortium nebulosum), we determined how annual variation in climate and density dependence explained the maintenance of two distinct morphs (terrestrial metamorph vs. aquatic paedomorph) in a high-elevation polyphenism. The effects of climate and conspecific density on morph development were evaluated with piecewise structural equation models (SEM) to tease apart the direct and indirect pathways by which these two mechanisms affect phenotypic plasticity. Climate had a direct effect on morph outcome whereby longer growing seasons favoured metamorphic outcomes. Also, climate had indirect effects on morph outcome as mediated through density-dependent effects, such as long overwintering coldspells corresponding to high cannibal densities and light snowpacks corresponding to high larval densities, both of which promoted paedomorphic outcomes. Both climate and density dependence serve as important proxies for growth and resource limitation, which are important underlying drivers of the phenotypic plasticity in animal polyphenisms. Our findings motivate new studies to determine how contemporary climate change will alter the selective pressures maintaining phenotypic plasticity and polyphenisms., (© 2024 The Author(s). Journal of Animal Ecology © 2024 British Ecological Society.)

We analyzed the expression of genes involved in the hypothalamic-pituitary-thyroid axis (HPT-axis) in the longfin yellowtail Seriola rivoliana early larva, including temperature effects (22, 26 and 28 °C) and days of development (day one, day two, and day six after hatching). We aimed to determine if egg and larval incubation at different temperatures could disrupt this critical endocrine axis, which, in an aquaculture context, it could provoke mortality during early metamorphosis. There was a significant interaction between temperature and developmental timing on the relative expression of thyrotropin releasing hormone (trh). Larvae at 22 °C was the longest and increased more trh expression than larvae at higher temperatures. Interestingly, thyrotropin stimulating hormone (tsh) was highly expressed after hatching. Subsequently, it was downregulated at any temperature at least until day four, suggesting a temporal inhibition of the HPT axis. Therefore, we suggest that tsh-binding (tshr) to follicles should have occurred from hatching, creating a further "cascade effect" of upregulation of larval thyroglobulin (tg) from day two in a temperature-dependent manner. Consequently, new thyroid hormones should have been produced after yolk sac absorption. The above may indicate a narrow window of larval survival, where the larval transition from endogenous to exogenous feeding would depend on the correct timing to synthesize tg. Temperature significantly affected the expressions of deiodinase 1 (dio1-downregulated) and deiodinase 2 (dio2-upregulated) after hatching. The expressions of thyroid receptors alpha (trα) and beta (trβ) remained constant after hatching without significant effects of temperature and days of development. Then, the differential expression on day six showed that all HPT-axis transcripts increased their expressions as larvae developed, which suggested a functional HPT. Finally, there was no evidence that any temperature would disrupt the endocrine's larval axis, which indicated that the longfin yellowtail has a wide temperature adaption. Nevertheless, based on tg upregulation, we suggest that larvae should be maintained around 25-26 °C after hatching for a better chance of survival and development., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Ocean acidity extreme (OAX) events, triggered by climate change and anthropogenic activities, are projected to become more intense and frequent in coastal ecosystems, devastating marine bivalves and ecosystems they support. Maternal effects adaptively modulate offspring performance in response to climatic stressors, but whether and to what extent they can confer offspring resistance to OAX remain largely unknown. Here, we investigated impacts of OAX on the parental and larval lipidomes of Manila clams (Ruditapes philippinarum) to add further insights into the energetic nature of maternal effects. A total of 177 significantly down-regulated lipid components (categorized into glycerolipids mainly) were shown in OAX-stressed adults compared with those reared under ambient conditions, and following parental conditioning, larvae also exhibited a further decreasing down-regulation of the glycerolipid components. Triacylglycerols were identified as the predominant composition of glycerolipids and the primary sources of energy for gonadal maturation and larvae development. Yet, larvae spawn from adults exposed to OAX had significantly lower contents of triacylglycerols than those without a prior history of parental conditioning, with the carbon chain length and unsaturation degree of the triacylglycerol components being significantly affected. The latter was also in line with significant increases in the production of triacylglycerol byproducts (diacylglycerols). Overall, our findings suggest that when OAX prevailed during reproductive seasons of Manila clams, maternal effects could be maladaptive by depressing the energetic deposition of larvae, and may not be a potential adaptive modulator of marine bivalves to cope with unprecedented environmental change., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)