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Hypoxia not only plays a critical role in multiple disease conditions; it also influences the growth and development of cells, tissues and organs. To identify novel hypoxia-related mechanisms involved in cell and tissue growth, studying a precise hypoxia-sensitive time window can be an effective approach. Drosophila melanogaster has been a useful model organism for studying a variety of conditions, and we focused in this study on the life cycle stages of Drosophila to investigate their hypoxia sensitivity. When normoxia-grown flies were treated with 4% O2 at the pupa stage for 3, 2 and 1 day/s, the eclosion rates were 6.1%, 66.7% and 96.4%, respectively, and, when 4% O2 was kept for the whole pupa stage, this regimen was lethal. Surprisingly, when our hypoxia-adapted flies who normally live in 4% O2 were treated with 4% O2 at the pupa stage, no fly eclosed. Within the pupa stage, the pupae at 2 and 3 days after pupae formation (APF), when treated for 2 days, demonstrated 12.5 ± 8.5% and 23.6 ± 1.6% eclosion, respectively, but this was completely lethal when treated for 3 days. We conclude that pupae, at 2 days APF and for a duration of a minimum of 2 days, were the most sensitive to hypoxia. Our data from our hypoxia-adapted flies clearly indicate that epigenetic factors play a critical role in pupa-stage hypoxia sensitivity.

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. [ABSTRACT FROM AUTHOR]

The compound eyes are crucial to honeybees, playing pivotal roles in color recognition, orientation, localization, and navigation processes. The development of compound eyes is primarily mastered by an evolutionarily conserved transcription factor Pax6. In honeybees, there are two Pax6 homologs: ey and toy. To gain a deeper understanding of their functions, we knock out both homologs using CRISPR/Cas9 technology. Intriguingly, we observe that toy knockout mutants have smaller heads without compound eyes and exhibit brain atrophy, while ey knockout mutants develop normal compound eyes, most of which die before/during their metamorphosis from pupa to adult. By comparing the head transcriptomes of four stages (larva, prepupa, pupa, and adult) in toy-knockout mutants versus normal controls, we identify significantly perturbed genes related to DNA binding transcription factors, neuron differentiation, and insect visual primordium development. Additionally, we find the interaction network of toy in honeybees differs obviously from that of D. melanogaster. Our findings suggest the two Pax6 genes serve distinct functions in honeybees and toy takes over the central function of ey in master-regulating the development of honeybee compound eyes. This adds new evidence for breaking the simplified view that some of conservative developmental toolkit genes function as all-or-nothing master regulators. Knockout of toy using CRISPR/Cas9 technology leads to small-headed honeybees without compound eyes, while ey-knockout bees have normal compound eyes, suggesting toy takes over the central role of ey in mastering honeybee compound-eye development. [ABSTRACT FROM AUTHOR]

Introduction: Considering the importance of comparing the silkworm performances of Iranian and imported commercial hybrids from China production traits, viability index and cocoon dimensions index were investigated in eight Iranian (31×32, 32×31, 103×104, 104×103, 151×154, 154×151, 153×154 and 154×153), and nine imported Chinese commercial hybrids (Suju×Minghu (S×M), Minghu×Suju (M×S), Baiyue×Qiufeng (B×Q), Qiufeng×Baiyue (Q×B), BaiyueB×QiufengA (BB×QA), QiufengA×BaiyueB (QA×BB), 781×7532, Lianggang No2 and Guican No 5) in the present study. Materials and Methods: After eggs hatched and according to the standard protocols, the studied hybrids were reared under the same conditions at Iran Silk Research Center (ISRC). Statistical analyses were done in three replications of 200 larvae (totally 600 records per each hybrid) based on completely randomized design. For comparing the means, Tukey's statistical test was used at a significance level of P<0.05. The GLM procedure was used in SAS software version 9.4. Data generated in respect of the studied traits were subjected for further analysis by a multiple trait evaluation index using the following formula: evaluation index (E.I.)= (A-B/C×10) + 50, where A is value of a particular breed for particular trait, B is mean value for a particular trait of all the hybrids, C is standard deviation of a particular trait for all the hybrids, 10 is standard unit and 50 is fixed value. Minimum / average Evaluation Index (E.I) value fixed for selection of a hybrid is >50. Based on the evaluation index (E.I) consisted of production traits and viability, best hybrids were identified. Results and Discussion: The result of the research showed the close competition between Iranian and Chinese silkworm hybrids. In the situation that the performance of Iranian 32×31 and 31×32 hybrids for traits related to weight, shell weight and shell percentage of best cocoons was higher than the other hybrids but based on a composite index that was used in this research, 151×154, 154×151, 153×154 and 154×153 hybrids showed superiority. Based on this index, M×S and S×M as well as BB×QA and QA×BB had a higher average than the other hybrids. This result is based on the average of two-way crosses for the hybrids that both pairs were used in the experiment. Conclusion: Generally, the results of mean comparisons and evaluation index showed that Iranian hybrids had better performance in all traits related to produce cocoons than the Chinese hybrids. Although, Chinese hybrids were significantly different from the Iranian for pupa viability percentage trait as Lianggang No2 hybrid showed the highest value. [ABSTRACT FROM AUTHOR]

Holometabolous insects undergo morphological remodeling from larvae to pupae and to adults with typical changes in the cuticle; however, the mechanism is unclear. Using the lepidopteran agricultural insect Helicoverpa armigera, cotton bollworm, as a model, we revealed that the transcription factor RUNT-like (encoded by Runt-like) regulates the development of the pupal cuticle via promoting a pupal cuticle protein gene (HaPcp) expression. The HaPcp was highly expressed in the epidermis and wing during metamorphosis and was found being involved in pupal cuticle development by RNA interference (RNAi) analysis in larvae. Runt-like was also strongly upregulated in the epidermis and wing during metamorphosis. Knockdown of Runt-like produced similar phenomena, a failure of abdomen yellow envelope and wing formation, to those following HaPcp knockdown. The insect molting hormone 20-hydroxyecdysonen (20E) upregulated HaPcp transcription via RUNT-like. 20E upregulated Runt-like transcription via nuclear receptor EcR and the transcription factor FOXO. Together, RUNT-like and HaPCP are involved in pupal cuticle development during metamorphosis under 20E regulation. Author summary: Holometabolous insects develop pupal cuticle during metamorphosis; however, the mechanism is unclear. Using the lepidopteran agricultural insect Helicoverpa armigera, cotton bollworm, as a model, we revealed that the transcription factor RUNT-like regulates a pupal cuticle protein gene, HaPcp, expression therefore regulates the development of the pupal cuticle under insect molting hormone 20-hydroxyecdysonen (20E) regulation. HaPcp is an insect specific gene without homologue in humans, and the insect Runt-like is different from vertebrate, too. This work deeply addresses the mechanism that 20E regulates development of insect pupal cuticle and presents new gene targets to explore the new growth regulators to control the pest. [ABSTRACT FROM AUTHOR]

Background: Black flies are among the most medically and veterinary important insects, as adult females of certain species are the sole vector of Onchocerca volvulus. Here, a new black fly species belonging to the subgenus Asiosimulium Takaoka & Choochote, 2005, is described and formally named as Simulium (Asiosimulium) kittipati sp. nov. Methods: Pupae and larvae of black flies were collected from available substrates in the stream from central Thailand. Pupae were individually separated in plastic tubes and maintained until adult flies emerged. The emerged adult flies associated with their pupal exuviae and cocoon as well as mature larvae preserved in 85% ethanol were used to describe the new species based on an integrated approach of morphological examination and molecular analysis of the COI gene. Results: The new species is characterized in the female by the medium-long sensory vesicle with a medium-sized opening apically, scutum with three faint longitudinal vittae, and the ellipsoidal spermatheca; in the male by the number of upper-eye (large) facets in 20 vertical columns and 21 horizontal rows, hind basitarsus slender, nearly parallel-sided, and median sclerite much wider and upturned apically; in the pupa by the head and thoracic integument densely covered with tiny tubercles, and the pupal gill of arborescent type with 28–30 filaments; and in the larva by the postgenal cleft deep, nearly reaching the posterior margin of the hypostoma, and dark pigmented sheath of the subesophageal ganglion. The DNA barcode successfully differentiated the new species from its congeners with an interspecific genetic divergence of 1.74–18.72%, confirming the morphological identification that the species is a new member of the subgenus Asiosimulium. Phylogenetic analyses also indicated that the new species is genetically closely related to Simulium phurueaense Tangkawanit, Wongpakam & Pramual, 2018, further supporting its morphological classification. Conclusions: This is the ninth species assigned to the subgenus Asiosimulium within the genus Simulium Latreille, 1802. Taxonomic notes and identification keys are given to distinguish this new species from the eight known species members in its same subgenus. Additionally, a distribution map of all species members in this subgenus occurring in Thailand and other countries is provided. [ABSTRACT FROM AUTHOR]

Winter diapause in insects is commonly terminated through cold exposure, which, like vernalization in plants, prevents development before spring arrives. Currently, quantitative understanding of the temperature dependence of diapause termination is limited, likely because diapause phenotypes are generally cryptic to human eyes. We introduce a methodology to tackle this challenge. By consecutively moving butterfly pupae of the species Pieris napi from several different cold conditions to 20 °C, we show that diapause termination proceeds as a temperature-dependent rate process, with maximal rates at relatively cold temperatures and low rates at warm and extremely cold temperatures. Further, we show that the resulting thermal reaction norm can predict P. napi diapause termination timing under variable temperatures. Last, we show that once diapause is terminated in P. napi, subsequent development follows a typical thermal performance curve, with a maximal development rate at around 31 °C and a minimum at around 2 °C. The sequence of these thermally distinct processes (diapause termination and postdiapause development) facilitates synchronous spring eclosion in nature; cold microclimates where diapause progresses quickly do not promote fast postdiapause development, allowing individuals in warmer winter microclimates to catch up, and vice versa. The unveiling of diapause termination as one temperature-dependent rate process among others promotes a parsimonious, quantitative, and predictive model, wherein winter diapause functions both as an adaptation against premature development during fall and winter and for synchrony in spring. [ABSTRACT FROM AUTHOR]

In this study, a powder of the larvae, pupae and adults of the fall armyworm Spodoptera frugiperda (J.E. Smith) was obtained, and 1% of this powder was added to solid culture media potato dextrose agar (PDA) and liquid media Sabouraud dextrose broth (SDB) and minimal medium (MM), to induce the development and virulence of the M1 strain of Metarhizium anisopliae (Metchnikoff) Sorokin, which is pathogenic against the L1 larvae of S. frugiperda. The pupa powder was selected because it causes the greatest degree of radial growth of the M1 strain (73.3 mm) in PDA (p < 0.05). The enzymatic activity of M1, in the medium with pupa powder of S. frugiperda (I), was increased on day 4 for the proteases in MMI (1.1 U min-1) and on day 2 for the lipases in MMI (7034 U) and SDBI (6004 U) (p<0.05). In the pathogenicity bioassay, the M1 strain grown on PDA with the pupa powder caused 90% mortality on day 4 in the L1 larvae, while M1 grown on PDA without the powder caused 50%. With the pupa powder, the growth of the M1 strain increased, which promoted the production of enzymes, thus, increasing the pathogenicity of M. anisopliae against the L1 larvae of S. frugiperda. [ABSTRACT FROM AUTHOR]

Simple Summary: Parasitoid wasps play a crucial role in controlling sanitary pests such as cockroaches, which pose a threat to human health. Tetrastichus hagenowii is an effective parasitoid wasp for the biological control of cockroaches, particularly the American cockroach (Periplaneta americana). This study aimed to evaluate the stability of potential reference genes in T. hagenowii. Seven candidate reference genes were rigorously selected and their reliability was assessed using RT-qPCR. Stability analysis across different developmental stages of T. hagenowii was conducted using five algorithmic methods, ultimately identifying α-tubulin as the most stable reference gene. Our research not only provides reliable reference genes to enhance studies on gene expression and functional genomics related to T. hagenowii but also offers valuable insights for the regulation of parasitoid wasps in cockroach control strategies. Parasitoid wasps play a crucial role in the efficient control of pests, a substantial menace to human health and well-being. Tetrastichus hagenowii (Ratzeburg) stands out as the most effective egg parasitoid wasp for controlling American cockroaches, but accurate and stable reference genes for quantitative real-time polymerase chain reaction of T. hagenowii genes are still lacking. In this study, we assessed seven candidate nuclear genes, including α-tubulin (α-TUB), elongation factor-1-alpha (EF-1α), β-actin (Actin), ribosomal protein 49 (RP49), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), nicotinamide adenine dinucleotide (NADH), and elongation factor 2 (EF2) of T. hagenowii. By analyzing expression stability with four algorithms (Delta Ct, geNorm, NormFinder, and BestKeeper), as well as comprehensive ranking with RefFinder, we identified α-TUB as the most stable reference gene for the larval, pupal, female adult, and male adult stages. Subsequently, we estimated the transcript levels of vitellogenin (Vg) and cuticle protein (CP) after normalization with α-TUB across various developmental stages. Significantly higher expression levels of CP and Vg were observed in pupae and female adults, respectively, consistent with previous findings in other insects. This study offers a reliable reference gene for normalizing transcription levels of T. hagenowii genes. [ABSTRACT FROM AUTHOR]

In the face of rising global temperatures, the mechanisms behind an organism's ability to acclimate to heat stress remain enigmatic. The rice leaf folder, Cnaphalocrocis medinalis, traditionally viewed as temperature‐sensitive, paradoxically exhibits robust larval acclimation to heat stress. This study used the heat‐acclimated strain HA39, developed through multigenerational exposure to 39°C during the larval stage, and the unacclimated strain HA27 reared at 27°C to unravel the transgenerational effects of heat acclimation and its regulatory mechanisms. Heat acclimation for larvae incurred a fitness cost in pupae when exposed to high temperature, yet a significant transgenerational effect surfaced, revealing heightened fitness benefit in pupae from HA39, even without additional heat exposure during larval recovery at 27°C. This transgenerational effect exhibited a short‐term memory, diminishing after two recovery generations. Moreover, the effect correlated with increased superoxide dismutase (SOD) enzyme activity and expression levels of oxidoreductase genes, representing physiological and molecular foundations of heat acclimation. Heat‐acclimated larvae displayed elevated DNA methylation levels, while pupae from HA39, in recovery generations, exhibited decreased methylation indicated by the upregulation of a demethylase gene and downregulation of two methyltransferase genes at high temperatures. In summary, heat acclimation induces DNA methylation, orchestrating heat‐stress memory and influencing the expression levels of oxidoreductase genes and SOD activity. Heat‐stress memory enhances the acclimation of the migratory insect pest to global warming. [ABSTRACT FROM AUTHOR]

Peckia (Peckia) chrysostoma (Wiedemann, 1830) (Diptera: Sarcophagidae) is a colonizer of cadavers in the Neotropical Region. Nevertheless, data on development for the P. (P.) chrysostoma (e.g., instar duration) and behavioral strategies used by the species for locating and colonizing a corpse are scant. We aimed to explore bionomic and reproductive aspects of the flesh fly P. (P.) chrysostoma, and in this article we: (a) provide quantitative data on the life cycle of P. (P.) chrysostoma; (b) present bionomic measurements (length and weight) of larvae and pupae; (c) describe intrauterine egg and larvae development; and (d) analyze the ovo/larviposition behavior by gravid females. Females showed ovaries with discernible eggs and larvae between 8 and 10 days (x̅ = 23.3 eggs/female). This study reports the first observation of egg deposition, an atypical behavior for the species. The average development time for immature stages was 22.24 h and 21.36 h for 1st and 2nd respectively, and 3rd showed an average development time of 80.47 h. Pupa had the longest duration (x̅ = 295.69 h). A direct increase was observed in weight (P < 0.05) and length (P < 0.05) throughout time. The average survival time of males and females is approximately 30 days. This study expands the knowledge on P. (P.) chrysostoma, such as facultative ovoviviparity under laboratory conditions and the life cycle, which may benefit future studies for accuracy in entomology-based estimation of minimum post-mortem interval (min PMI). [ABSTRACT FROM AUTHOR]

Sarcophaga (=Parasarcophaga) (Liopygia) ruficornis (Fabricius, 1794) is a species of medical–veterinary and forensic importance, as its immatures cause myiasis in humans and animals and colonize carcasses and cadavers. Therefore, investigations into the biology and morphology of this species, with a particular focus on pupae that constitute ≥50% of the developmental period for the immatures, are pertinent. Although there are biological and morphological studies of pupae, the intrapuparial development at different temperatures has not yet been analysed. Therefore, the present study aimed to describe how temperature (22, 27 and 32 ± 1°C) affects the development and morphology of S. (L.) ruficornis pupae at 60 ± 10% relative humidity and a 12‐h photoperiod. Ten pupae were collected, euthanized and fixed every 4 h from pupariation until 24 h and every 8 h until the emergence of the first adult. Emergence occurred at 440, 272 and 232 h at 22, 27 and 32°C, with 590, 380 and 330 pupae, respectively. The highest mortality occurred at 32°C. Eight periods were defined, and sex was determined in pharate adult stage; in addition, 40 key morphological characteristics to estimate pupal age were presented. These findings can serve as a basis for studies on the biology and morphology of the pupa of S. (L.) ruficornis, particularly in bionomics, control and forensics, helping researchers and experts. [ABSTRACT FROM AUTHOR]

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In the current study, the effect of lethal concentrations of drug solutions and chemicals against pupae of Culex pipiens molestus mosquitoes was investigated after 24 hours and up to 3 days of the experiment using different concentrations. Regarding the drug solutions, the concentration of 100 ppm of the niclosamide drug solution showed a 100% lethal effect on the second day of the experiment, and the value of LC50 was 113.7 ppm. The concentration of 800 ppm of the sulfasalazine drug solution caused the death of all the pupa, with a death rate of 100% on the third day of the experiment, and the value of LC50 was 854.7 ppm. The concentration of 2000 ppm for both piperazine and isoniazid medicinal solutions caused a death rate of 100% on the second and third day of the experiment, respectively, and the LC50 value for both piperazine and isoniazid was 2220.7 and 2273.0 ppm, respectively. The concentration of 14,000 ppm of the drug solution 4-aminoantipyrine caused the death of all pupal mosquitoes on the second day of the experiment, and the value of LC50 was 14,360.0 ppm. As for solutions of chemicals, the concentration of 150 ppm of the chemical solution ethyl salicylate showed a lethal effect on all pupal mosquitoes, with a 100% death rate on the second day of the experiment, and the value of LC50 was 176.2 ppm. The death rate of mosquito pups reached 100% when using chemical solutions of thymol, sulfamic acid, sulfanilamide, and 2,6-dihydroxytoluene at concentrations of 240, 2000, 2500 and 10,000 ppm, respectively. The value of LC50 for each of thymol and sulfamic acid was, Sulfanilamide and 2,6-dihydroxytoluene 352.1, 2940.8, 2582.5 and 10750.0 ppm, respectively.

Abstract Background Culex pipiens L. is a principal vector of zoonotic arboviruses in Europe, acting in both an amplification role in enzootic transmission between avian hosts and as a bridge vector between avian hosts and mammals. The species consists of two forms which are indistinguishable using morphological methods but possess varying ecological and physiological traits that influence their vector capacity. In this study we validate methods that can be used to extract trace DNA from single pupal exuviae of Cx. pipiens for use in molecular speciation of samples. These DNA extraction methods are compared using measurement of the total yield and successful identification using a real-time polymerase chain reaction (PCR) assay. Results Genomic DNA was initially extracted from colony-derived individuals using an ethanol precipitation method, two commercially available DNA extraction kits: DNeasy® Blood & Tissue Kit (Qiagen, UK) and Wizard® SV Genomic DNA Purification System (Promega, UK) and a direct real-time PCR method. Time elapsed between eclosion and processing of pupae significantly influenced Cx. pipiens form identification as nucleic acid concentration and PCR amplification success decreased with increased time elapsed. Real-time PCR amplification success, however, was not shown to vary significantly between the three extraction methods, with all methods successfully identifying all samples, but the direct real-time PCR method achieved a lesser amplification success rate of 70% (n = 20 for each treatment). More variable results were produced when field-derived exuviae were used, with no significant difference in real-time PCR amplification success found across the four methods and a lower overall rate of successful identification of 55–80%. Conclusions This study shows that both colony and field derived Cx. pipiens pupal exuviae can be a useful non-invasive source of trace DNA permitting accurate biotype differentiation for at least twenty-four hours post-eclosion. The significance and utility of this technique in ecological and behavioural studies of Cx. pipiens is discussed and recommendations made for use according to experimental scenario.

Black soldier flies (Hermetia illucens (Linnaeus)) or BSF are insects that process organic waste, with larvae rich in nutrients and potential as an alternative protein source for food and feed. Astaxanthin, an antioxidant found in plants, algae, and crustaceans, is also assumed to be present in insects like BSF. This study examined the impact of feeding different organic wastes on BSF larval growth and astaxanthin content in its pupae. The experimental design involved four treatments: fruit and vegetable scraps (BS), tofu pulp (AT), rice leftovers (N), and a mix of fruit and vegetable scraps, tofu pulp and rice left-over (C), with animal feed T51 as control. Each treatment was repeated three times. Growth parameters of BSF such as weight, length, and larval characteristics, along with the chemical content of pupae, including astaxanthin, were measured. Pupae were extracted using a triphasic maceration method with n-heptane, ethyl acetate, acetonitrile, butanol-l, and distilled water. Compound identification was done using terpenoid qualitative tests, thin layer chromatography (TLC), and liquid chromatography-mass spectrometry (LC-MS). The findings revealed that fruit and vegetable scraps (BS) significantly increased larval body size, with a weight of 0.09 ± 0.04 grams/larvae and a length of 1.49 ± 0.31 cm/larvae. LC-MS analysis detected lutein derivatives, anisodamine, resveratrol derivatives, dehydroarginine, and astaxanthin derivatives with a molecular mass of 597.23 m/z in BSF larvae fed on fruit and vegetable scraps.

The three-dimensional morphologies of insects, such as spines and horns, have recently garnered attention as an effective system for elucidating the processes underlying dramatic changes in external morphology. Adult leaf beetles of Dactylispa Weise 1897 exhibit sharp spines on their pronotum and elytra, while their pupae lack such features. In order to obtain foundational data for future investigations into the developmental mechanisms governing spine formation, this study assessed variations in spine numbers across ten body regions and in spine size across three body regions in adults of Dactylispa higoniae (Lewis in Ann Mag Nat Hist (Ser. 6) 17:329–343, 1896. https://doi.org/10.1080/00222939608680376) and D. issikii (Chûjô in Bull Umeno Entomol Lab 6:5–13, 1938). As a result, the degree of variation in spine numbers and size was variable among body regions even within single species. However, the number of spines on the pronotal front margin in D. higoniae and the elytral interval 4 in D. issikii was stable. The spine numbers of these two species did not exhibit significant differences between the right and left sides of the body and was not likely to be influenced by sex or elytral length. Furthermore, we observed pronotal and elytral spines that formed inside the pupal cuticle of late-stage pupae of these two species. Our observations suggested neither pronotal nor elytral spines exhibit a formation pattern resembling concentric folded structures, as seen in beetle horns. [ABSTRACT FROM AUTHOR]

Summary: The purpose of this experiment was to explore the improvement of tussah pupa powder on the deterioration of pork batter emulsion gel by using proteomics, distribution of moisture, rheological properties and surface hydrophobicity. The results showed that the myosin decomposed as the heating temperature increased; as the increase of the addition proportion of tussah pupa powder at each sterilisation temperature point, compared to the control group, the pH of a pork batter emulsified gel showed a significant increased trend (P < 0.05); the low molecular weight proteins or formed oligomers of tussah pupa powder could be used as fillers to participate in the formation process of gel network; the vitellogenin of tussah pupa powder could form a 'skeleton' in the pork batter system; as the sterilisation temperature at 100 °C (30 min) and 110 °C (30 min), the surface hydrophobicity was the largest as the addition amount 2%. In short, the tussah pupae powder could improve the deterioration characteristics of the pork batter emulsified gel by influencing the pH, surface hydrophobicity and the formation of gel network of pork batter system. This research provided theoretical support for improving the process property of the meat products. [ABSTRACT FROM AUTHOR]

Entomopathogenic fungi are the most effective control remedy against a wide range of medical and agricultural important pests. The present study aimed to isolate, identify, and assess the virulence of Metarhizium rileyi against Spodoptera litura and Spodoptera frugiperda pupae under soil conditions. The biotechnological methods were used to identify the isolate as M. rileyi. The fungal conidial pathogenicity (2.0 × 107, 2.0 × 108, 2.0 × 109, 2.0 × 1010, and 2.0 × 1011 conidia/mL−1) was tested against prepupae of S. litura and S. frugiperda at 3, 6, 9, and 12 days after treatments. Additionally, the artificial soil‐conidial assay was performed on a nontarget species earthworm Eudrilus eugeniae, using M. rileyi conidia. The present results showed that the M. rileyi caused significant mortality rates in S. litura pupae (61‐90%), and S. litura pupae were more susceptible than S. frugiperda pupae (46%–73%) at 12 day posttreatment. The LC50 and LC90 of M. rileyi against S. litura, were 3.4 × 1014–9.9 × 1017 conidia/mL−1 and 6.6 × 105–4.6 × 1014 conidia/mL−1 in S. frugiperda, respectively. The conidia of M. rileyi did not exhibit any sublethal effect on the adult stage of E. eugeniae, and Artemia salina following a 12‐day treatment period. Moreover, in the histopathological evaluation no discernible harm was observed in the gut tissues of E. eugeniae, including the lumen and epithelial cells, as well as the muscles, setae, nucleus, mitochondria, and coelom. The present findings provide clear evidence that M. rileyi fungal conidia can be used as the foundation for the development of effective bio‐insecticides to combat the pupae of S. litura and S. frugiperda agricultural pests. [ABSTRACT FROM AUTHOR]

Osmia excavata is an excellent pollinator in nature and plays a vital role in the conservation of agro-ecosystems and food security. Given the important role of the gut bacterial community in host health and regulation of host growth and development, using 16S rRNA gene sequencing data, the present study explored the composition of the gut bacterial community and its diversity at different life stages (eggs, young larvae, old larvae, young pupae, old pupae, and 1-day-old adults in cocoons) of the solitary bee Osmia excavata. The results showed that the core phyla in the gut of O. excavata at different life stages were Proteobacteria, Firmicutes, Bacteroidota, and Actinobacteriota, and the core genera were Sodalis, Tyzzerella, and Ralstonia. The highest intestinal bacterial diversity was found in the Egg period, and the lowest bacterial alpha diversity was found in the 1-day-old Adult period; the bacterial diversity of O. excavata showed a process of decreasing, as it was growing from the Egg period to the 1-day-old Adult period. Our study found that O. excavata undergoes a significant change in the structure of the gut flora when it grows from the young pupae to old pupae stage, a period of growth that coincides with the process of cocooning and isolation from the external environment after food depletion. This suggests that food and environmental factors are key contributors to the structure of the bacterial community in the gut of solitary bees. [ABSTRACT FROM AUTHOR]

BACKGROUND: Actin‐related protein 2/3 complex regulates actin polymerization and the formation of branched actin networks. However, the function and evolutionary relationship of this complex subunit 2 (Arpc2) has been poorly understood in insects. RESULTS: To address these issues, we performed comprehensive analysis of Arpc2 in Tribolium castaneum. Phylogenetic analysis revealed that Arpc2 was originated from one ancestral gene in animals but evolved independently between vertebrates and insects after species differentiation. T. castaneum Arpc2 has a 906‐bp coding sequence and consists of 4 exons. Arpc2 transcripts were abundantly detected in embryos and pupae but less so in larvae and adults, while it had high expression in the gut, fat body and head but low expression in the epidermis of late‐stage larvae. Knockdown of it at the late larval stage inhibited the pupation and resulted in arrested larvae. Silencing it in 1‐day pupae impaired eclosion, which caused adult wings to fail to close. Injection of Arpc2 dsRNAs into 5‐day pupae made adults have smaller testis and ovary and could not lay eggs. The expression of vitellogenin 1 (Vg1), Vg2 and Vg receptor (VgR) was downregulated after knocking down Arpc2 5 days post‐adult emergence. Arpc2 silencing reduced 20‐hydroxyecdysone titer by affecting the enzymes of its biosynthesis and catabolism but increased juvenile biosynthesis via upregulating JHAMT3 expression. CONCLUSION: Our results indicate that Arpc2 is associated with the metamorphosis and reproduction by integrating ecdysone and juvenile hormone metabolism in T. castaneum. This study provides theoretical basis for developing Arpc2 as a potential RNA interference target for pest control. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Background: The fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is an aggressive pest species that causes severe economic losses in outbreak regions. Use of FAW natural enemies, particularly native parasitoids, has been suggested as a promising control strategy. Campoletis chlorideae Uchida (Hymenoptera: Ichneumonidae) is a solitary ichneumonid larval endoparasitoid with a broad host range that includes S. frugiperda. However, its parasitism rate, developmental stages, and population dynamics during parasitization of S. frugiperda remain unclear. A field survey was conducted to determine the emergence rate and sex ratio of C. chlorideae, evaluated the biological control performance, and investigated the production potential of it on the individually reared and group-reared S. frugiperda through age-stage, two-sex life tables, respectively. Results: The results showed that C. chlorideae parasitizing individually reared FAW resulted for the parasitoid in a lifetime total fecundity (F) of 301.5 ± 16.4 eggs/female, a net reproduction rate (R0) of 62.03 ± 9.07 adult females/female, an overall life span of females of 28.3 ± 0.52 days, and an intrinsic rate of increase (r) of 0.1946 ± 0.0076 day−1. In contrast, the parameters of F, R0, overall life span of females, and r were 87.71 ± 6.32 pupae/female, 6.02 ± 1.61 adult females/female, 25.21 ± 0.79 days, and 0.0918 ± 0.0148 day−1 in group-reared S. frugiperda. Conclusions: This study suggests that C. chlorideae has a promising biological control potential against S. frugiperda. Future research should focus on developing methods to increase the production of C. chlorideae. [ABSTRACT FROM AUTHOR]

Bombyx mori is a lepidopteran holometabolous insect with distinct developmental stages: egg, larvae, pupae, and adult. The lepidopteran insect undergoes major modifications in the central nervous system (CNS) so as to adapt to the lifestyle of these distinct stages with specific habitats and functions from voraciously feeding larval stages to flying reproductive adults via dormant pupal stages. Such transitions are linked to transcriptional, epigenetic, and translational complexities. Therefore, studying rhythmic gene expression in CNS of various developmental stages and the effects of antagonists on developmental hormones requires a very stable reference gene (RG). To facilitate rhythmic gene expression studies using reverse transcription quantitative polymerase chain reaction (RT-qPCR) in B. mori and the effect of developmental hormone juvenile hormone (JH) and 20-hydroxy ecdysone hormone (20 HE), antagonists Precocene 1 and testosterone, respectively, were used. Eight candidate RGs, namely, Translational initiation factor 3 subunit 4 (TI3S4), Translational initiation factor 3 subunit 5 (TI3S5), Ribosomal protein subunit 7 (RPs7), TATA-binding protein association factor (TAF13), Translational initiation factor 4 A (TI4A), Ribosomal protein (RPL32), Elongation factor 1 (EF1), and Arginine kinase (AK), were assessed in the CNS of B. mori. The postembryonic developmental (PED) stages used were the fifth late larval instar, early pupa, mid pupa, late pupa, and adult. The assessments were done at four different time points, Zeitgeber time (ZT) 0, 6, 12, and 18, to find stability towards 24-h rhythmic expression. RefFinder, geNorm, and Ct value analysis were performed. RefFinder and geNORM studies suggested stability order as TI3S4 > TI3S5 > RPs7, but Ct value evaluation showed stability order as TI3S5 > TI3S4 > RPs7. We therefore demonstrated that TI3S4, TI3S5, and RPs7 can be used as RG in various PED stages in CNS of B. mori (Strain: CB-hybrid, PMÃ--CSR2) towards studies with effects of JH and 20 HE antagonists. [ABSTRACT FROM AUTHOR]

The study of immature stages of the Lycidae family is currently limited. Only approximately 2% of the described species have documented larvae in different instars and pupae, with a focus mainly on the fauna of the Old World. During fieldwork in Hidalgo, Mexico, we collected adult pairs of Neolycus fernandezi Dugès 1878 and Rhyncheros simulans. We kept the adults in laboratory conditions and obtained the eggs and first instar larvae. The comparison of chaetotaxies between N. fernandezi and R. simulans provides useful characters for differentiating genera. Additionally, the morphological characters of the first larval were compared with those described for Lycostomus ferrugineus Fabricius, 1798, L. formosanus Pic, 1937, as well as Calopteron terminale (Say, 1823). These comparisons revealed differences in the shape of the epicranium, relationship between the length of the maxillary palpomere and mala, prementum division, prosternum amplitude, pronotum shape, and abdominal spiracular plate division. The knowledge of the immature stages of two species within the Lycini tribe is significant since the egg and first instar larval of any American species belonging to this tribe have yet to be identified. [ABSTRACT FROM AUTHOR]

Piercing-sucking insects are extremely efficient vectors of animal and plant pathogens. This group is polyphyletic with the piercing organ highly heterogenous in its structure and morphogenesis, adapted to a specific host and feeding mechanism. For instance, hemimetabolous aphids feed on plant sap from nymphs to adults and they renew their stylets at each molt, thanks to specialized secreting glands. Distinctly, holometabolous mosquitoes have two feeding modes: larvae feed in water by filtering and scraping surfaces with their mouths, while female adults can acquire blood from vertebrate hosts. The pupal metamorphosis allows switching from one feeding habit to another. Here, we present a deep characterization of the biogenesis of the adult mouthparts in parthenogenetic females of the pea aphid Acyrthosiphon pisum (Harris 1776) (Hemiptera: Aphididae) and in females of the mosquito Aedes albopictus (Skuse 1895) (Diptera: Culicidae), investigated using non-invasive X-ray synchrotron-based microtomography. Comparing datasets collected from aphid juvenile and adult stages and from preimaginal and adult stages of the mosquito, we were able to track the morphological changes of secreting glands and the synthesis of the adult stylet in aphid heads and to follow the de novo formation of mosquito mouthparts in pupae. Our study provides a baseline for investigating the evolution and the development of piercing-sucking mouthparts and to better understand how morphogenesis works in insects. [ABSTRACT FROM AUTHOR]

The species formerly known as Scrobipalpa pauperella (Heinemann, 1870) is hereby revised to Scrobipalpa freyii (Scott, 1859). An account is given of the discovery in the British Isles of the larva mining the leaves of Centaurea scabiosa L. (Asteraceae). The larva, pupa, adult and genitalia of both sexes are compared with those of Scrobipalpa acuminatella (Sircom, 1850). Images are provided of both species. [ABSTRACT FROM AUTHOR]

A common management intervention to support declining wild pollinators is 'pollinator planting'. However, despite years of inclusion in conservation initiatives, global pollinator declines continue.Using the agent‐based model BEE‐STEWARD with two example species, Bombus terrestris (L. 1758) and B. pascuorum (Scopoli, 1763) (Hymenoptera: Apidae), we explore when during the year bumblebee resource demand is the highest and how that relates to seasonal changes in colony composition (numbers of eggs, larvae, pupae and adults). We then examine the impact of two‐week periods of resource scarcity across the year. Finally, we explore how enhancement with early spring‐blooming herbaceous species or trees changes colony survival and queen production.In the United Kingdom, there is a previously under‐appreciated 'hungry gap' for bumblebees in March–April, before the peak flight period, driven by the demands of larvae for protein and thermoregulation in the colony, rather than the number of adult bees.A 2‐week gap in forage availability during this period drives a 50%–87% drop in the production of daughter queens. Adding early‐blooming species in the model had significant, positive, long‐term effects on colony survival probability and daughter queen production.Pollinator‐planting initiatives in both national and international conservation schemes need to include plants that flower up to 1 month before the adults of target social pollinator species are apparent in the field, during the period that larvae dominate the colony. This approach is likely to increase colony survival and queen production, contributing towards halting and reversing global pollinator decline. [ABSTRACT FROM AUTHOR]

The tomato leafminer, Tuta absoluta is a devastating invasive pest that poses a serious threat to tomato crops worldwide. Its extensive global dispersion serves its capacity to adapt to variations in climate conditions. In this context, the pupa is the most resistant stage to prolonged exposure to cold temperatures. Therefore, indicators of cold resistance were studied in overwintering pupae collected from the field and pupae reared under two constant conditions, high temperature, and long day (25°C, 16:8 L:D and 65±5% RH), and low temperature and short day (15°C, 13:11 L:D and 65±5% RH). The results show that the super cooling point (SCP) significantly decreased in December (-20.5±1.2°C) and January (-20.26±0.78°C) with a decrease in temperature. In the laboratory, the decrease in temperature and photoperiod increased the tolerance of pupae to subzero temperatures. Lethal Temperature 50 (LT50) and LT90 of pupae collected in the field were recorded at -13.70 to -10.23°C and -18.73 to -15.37°C, respectively. A comparison of lethal temperatures with the lowest ambient temperature in December and January indicated that T. absoluta has a high overwintering potential in Karaj, Alborz Province, Iran, and can easily survive cold winters. [ABSTRACT FROM AUTHOR]

BACKGROUND: Silkworm (Bombyx moil L.) Pupa protein (SPP) is a high‐quality insect protein and is considered a sustainable alternative source for traditional animal food protein. However, the utilization of SPP is limited because of its low solubility and emulsifying ability. In the present study, the synergistic effect of hydration and pulsed ultrasound on the physicochemical properties of SPP and SPP‐stabilized Pickering emulsions was evaluated. RESULTS: Pulsed ultrasound changed the particle size of SPP and its conformation. As the pulsed ultrasound increased from 0 s to 5 s, the α‐helix and SS contents of SPP decreased, whereas the β‐sheet and SH contents increased, which in turn improved its solubility and amphiphilicity. As a result, the SPP treated by a combination of 12 h of hydration and 3 s of ultrasound exhibited a contact angle of 74.95°, hydrophobicity of 904.83, EAI of 6.66 m2 g−1 and ESI of 190.69 min. Compared with the combination of 1 h of hydration and 5 s of ultrasound, the combination of 12 h of hydration and 3 s of ultrasound exerted more soluble and hydrophobic SPP, whereas the EAI and ESI of the samples were higher. Notably, the ultrasound‐treated SPP can form a stable gel‐like emulsion (oil fraction ranging from 70% to 80%). CONCLUSION: The combination of hydration and ultrasound can effectively improve the physicochemical characteristics of SPP as well as its emulsion stability. Sufficient hydration is a cost‐effective method for facilitating the modification of proteins by ultrasound treatment. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Tenebrio molitor gained recognition for its nutritional value for human and/or animal consumption, its utilization as a model species in research studies, and its ability to degrade many types of plastics. However, it is a stored-product pest infesting a wide range of commodities in storages. The impact of temperature on the development of T. molitor was evaluated, taking into account confounding effects of other covariates such as developmental stage and larval instar. The time for larval development was longer at 22.5 °C than at 25 °C, 27.5 °C, and 30 °C. Tenebrio molitor spent most of its lifetime as larva > egg, pupa, since the outcome of our model inference resulted that the estimated probability of longer duration in larvae is approximately 25.5 times higher compared to eggs and pupae, and passed through 26 instars before it became pupa. Duration of larval instars increased significantly up to L22 and then decreased until L26. Among larval instars, L22 exhibited the highest developmental duration while the lowest was recorded for L1. These findings enhance the knowledge about the biology of T. molitor, leading to precise decisions for its successful culture. [ABSTRACT FROM AUTHOR]

The comparative studies under laboratory conditions have been carried out to investigate the influence of different alternative host plants viz., bael [Aegle marmelos (L.)], curry leaves [Murraya koenigii (L.) Spreng.], ber (Zizyphus mauritiana Lam.) and lemon [Citrus limon (L.) Burm.f.] on biology and morphometrics of Lemon butterfly, Papilio demoleus (Linn.). The lemon is the most preferred host of P. demoleus used as a controlled experimental host plant. The variations in periods of larval, pupal and rate of emergence into adult have been recorded and found that the lemon was the most preferred host plant on which all the larval stages survived smoothly. The caterpillars when reared on alternative hosts showed an increase in average larval period. The average survival percent of larvae were reported highest on bael leaves, followed by curry leaves and ber leaves. Similar patterns of results were also recorded on the pupal period and rate of emergence with alternative hosts. The morphometric variations in weight, size and colour of caterpillars, pupae and adults were also measured and found in decreasing order of host preferences as lemon>bael>curry>ber. [ABSTRACT FROM AUTHOR]

Silkworm (Bombyx mori) pupae are popular edible insects with high nutritional and therapeutic value. Currently, there is growing interest in the comprehensive application of silkworm pupae. In this study, peptides that exhibited anti-photoaging activity were obtained from silkworm pupae protein, aiming to investigate the protective effects and potential mechanisms of silkworm pupae peptides (SPPs) on skin photoaging. The results showed that SPPs were composed of 900 short peptides and could effectively alleviate skin photoaging progression. They significantly eliminated excessive production of ROS and MDA; meanwhile, they also renovated the antioxidant enzyme activities. The biomarkers related to collagen synthesis and degradation, including hydroxyproline, interstitial collagenase, and gelatinase, demonstrated that SPPs could suppress collagen degradation. Histopathological results showed that SPPs could reduce the inflammatory infiltrate and the thickness of the dermis and epidermis, as well as increase the collagen bundles and muscle fibers. The histopathological and biochemical results confirmed that SPPs could alleviate photoaging by inhibiting abnormal skin changes, reducing oxidative stress, and immune suppression. Overall, these data prove the protective effects of SPPs against the photoaging process, suggesting their potential as an active ingredient in skin photoaging prevention and therapy. [ABSTRACT FROM AUTHOR]

BACKGROUND: Galeruca daurica has become a new pest on the Inner Mongolia grasslands since an abrupt outbreak in 2009 caused serious damage. As a pupa indicator during insect metamorphosis, the early response gene of the ecdysone signaling pathway, Broad‐Complex (Br‐C), plays a vital role in the growth and development of insects. MicroRNAs (miRNAs) are small non‐coding RNAs which mediate various biological activities, but it is unknown whether and how Br‐C is regulated by miRNAs. RESULTS: Temporal expression profiles revealed that miR‐285 and Br‐C basically displayed an opposite trend during larval–adult development, and Br‐C was sharply up‐regulated on the last day of final‐instar larvae while miR‐285 was significantly down‐regulated. Both dual‐luciferase reporter assay and miRNA–mRNA interaction assay indicated that miR‐285 interacts with the coding sequence of Br‐C and represses its expression. Not only overexpression but also downexpression of miR‐285 led to the failure of larval to pupal to adult metamorphosis. In addition, both overexpression of miR‐285 and silence of Br‐C inhibited the expression of Br‐C and other ecdysone signaling pathway genes, including E74, E75, ECR, FTZ‐F1, and HR3. On the contrary, suppressing miR‐285 obtained opposite results. Further experiments showed that 20‐hydroxyecdysone down‐regulated miR‐285 and up‐regulated Br‐C and above‐mentioned genes, whereas juvenile hormone alalogue (JHA) resulted in opposite effects. CONCLUSION: Our results reveal that miR‐285 is involved in mediating the metamorphosis in G. daurica by targeting Br‐C in the ecdysone signaling pathway. miR‐285 and its target Br‐C could be as a potential target for G. daurica management. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Background: Culex pipiens L. is a principal vector of zoonotic arboviruses in Europe, acting in both an amplification role in enzootic transmission between avian hosts and as a bridge vector between avian hosts and mammals. The species consists of two forms which are indistinguishable using morphological methods but possess varying ecological and physiological traits that influence their vector capacity. In this study we validate methods that can be used to extract trace DNA from single pupal exuviae of Cx. pipiens for use in molecular speciation of samples. These DNA extraction methods are compared using measurement of the total yield and successful identification using a real-time polymerase chain reaction (PCR) assay. Results: Genomic DNA was initially extracted from colony-derived individuals using an ethanol precipitation method, two commercially available DNA extraction kits: DNeasy® Blood & Tissue Kit (Qiagen, UK) and Wizard® SV Genomic DNA Purification System (Promega, UK) and a direct real-time PCR method. Time elapsed between eclosion and processing of pupae significantly influenced Cx. pipiens form identification as nucleic acid concentration and PCR amplification success decreased with increased time elapsed. Real-time PCR amplification success, however, was not shown to vary significantly between the three extraction methods, with all methods successfully identifying all samples, but the direct real-time PCR method achieved a lesser amplification success rate of 70% (n = 20 for each treatment). More variable results were produced when field-derived exuviae were used, with no significant difference in real-time PCR amplification success found across the four methods and a lower overall rate of successful identification of 55–80%. Conclusions: This study shows that both colony and field derived Cx. pipiens pupal exuviae can be a useful non-invasive source of trace DNA permitting accurate biotype differentiation for at least twenty-four hours post-eclosion. The significance and utility of this technique in ecological and behavioural studies of Cx. pipiens is discussed and recommendations made for use according to experimental scenario. [ABSTRACT FROM AUTHOR]

During the winter, animals face limited food availability. Many animals enter dormancy to reduce their winter energy expenditure. Most insects spend the winter in diapause, a state of programmed dormancy. It is often assumed that diapausing insects need nutrient stores to fuel their many months of basal metabolism and must grow heavier than their non‐diapause‐programmed counterparts. However, the extent to which food limitation affects body weight during overwintering preparation as well as the likelihood and duration of diapause remains unclear. We limited the duration of the feeding period and thus the total quantity of food available to diapause‐destined larvae of the pupal‐diapausing flesh fly, Sarcophaga crassipalpis, to test how food limitation affects body weight in the context of diapause programming. We also tested the extent to which food deprivation and body weight affect the likelihood and duration of diapause. We hypothesized that diapause‐destined larvae grow more quickly and pupariate at a heavier body weight than non‐diapause larvae. We also hypothesized that body weight is more dramatically reduced by food limitations when a larva is programmed for diapause. Finally, we hypothesized that larvae with lighter body weight (i.e., food limited) are less likely to enter pupal diapause and also stay in diapause for a shorter duration than heavier, well‐fed, individuals. Contrary to our hypotheses that diapausing insects are heavier than their non‐diapausing counterparts, we found diapausing pupae weighed less than non‐diapausing pupae, especially when larvae received limited food. We found light pupae did not abort their diapause program. In both diapausing and non‐diapausing pupae, body weight was positively correlated with simulated winter survival. However, above a weight threshold, body weight no longer affected simulated winter survival in diapausing pupae. Contrary to our predictions and the general consensus in much of the diapause literature, we also found that lighter pupae stayed in diapause longer than heavier pupae. Overall, our results challenge the precept that body weight and diapause are positively associated. The relationship between body weight and diapause is complex and may be affected by the availability of food before and after winter, the availability of high‐quality overwintering sites, and the life history of a particular insect. [ABSTRACT FROM AUTHOR]

Monarch butterflies, Danaus plexippus L., in western North America are undergoing a seemingly volatile shift in seasonal population dynamics which includes novel winter-breeding on non-native milkweed in urban areas of northern California. The survival, viability and outcome of monarch pupae in the wild was evaluated for the first time by regular monitoring of a cohort of 104 pupae in a San Francisco managed urban landscape during the winter of 2021/22. Seventeen pupae were destroyed by landscaping activities. Almost half (49.4%) of the remaining pupae eclosed, with ∼ 70% likely infected with varying levels of the protozoan parasite Ophryocystis elektroscirrha (OE). A third of the pupae (32.2%) died of unknown causes, while the remainder (18.4%) disappeared, presumed predated. Two monarchs were killed while eclosing, by European paper wasps, Polistes dominula (Christ, 1791). Microscopic examination of pupal exuviae for spores of OE appeared to be a satisfactory proxy for microscopic examination of butterfly abdomens, in determining the presence/absence of infection. The developing importance of winter breeding in western monarch ecology necessitates modifications to conservation guidelines that enhance the survival of immature stages, acknowledging the role of non-native milkweeds while minimizing parasite infection. [ABSTRACT FROM AUTHOR]

In this study, we pioneered an alternative technology for manufacturing subunit influenza hemagglutinin (HA)-based vaccines. This innovative method involves harnessing the pupae of the Lepidoptera Trichoplusia ni (T. ni) as natural biofactories in combination with baculovirus vectors (using CrisBio® technology). We engineered recombinant baculoviruses encoding two versions of the HA protein (trimeric or monomeric) derived from a pandemic avian H7N1 virus A strain (A/chicken/Italy/5093/99). These were then used to infect T. ni pupae, resulting in the production of the desired recombinant antigens. The obtained HA proteins were purified using affinity chromatography, consistently yielding approximately 75 mg/L of insect extract. The vaccine antigen effectively immunized poultry, which were subsequently challenged with a virulent H7N1 avian influenza virus. Following infection, all vaccinated animals survived without displaying any clinical symptoms, while none of the mock-vaccinated control animals survived. The CrisBio®-derived antigens induced high titers of HA-specific antibodies in the vaccinated poultry, demonstrating hemagglutination inhibition activity against avian H7N1 and human H7N9 viruses. These results suggest that the CrisBio® technology platform has the potential to address major industry challenges associated with producing recombinant influenza subunit vaccines, such as enhancing production yields, scalability, and the speed of development, facilitating the global deployment of highly effective influenza vaccines. [ABSTRACT FROM AUTHOR]

For decades, the honeybee, Apis mellifera, has suffered from severe colony losses due to the ectoparasitic mite Varroa destructor. Various strategies based on chemicals fail to adequately control varroa mite populations, and often comprise side-effects on the host, parasite resistance and residues in hive products. Reduced temperature tolerance of V. destructor compared to its host has long been recognised and accordingly, the potential of hyperthermia to disrupt mite reproduction within honeybee brood cells or even kill adult parasites. Yet, earlier studies on hyperthermia remain largely anecdotal, and readily implementable solutions have so far been lacking. This study investigates autonomously controlled interval heating from within brood combs throughout the season compared to control colonies maintained according to good apicultural practice. We documented treatment-dependent colony growth dynamics and honey production at three apiaries, complemented by regular monitoring of varroa mite levels and comprehensive digital assessments of brood development over time. Our one-year field trial suggests the evaluated hyperthermia device efficiently suppresses mite populations below critical thresholds until autumn. Whilst a general winter reference-curing revealed similar mite infestations of colonies previously treated with hyperthermia versus formic acid (control), only the latter imposed substantial overwintering burdens indirectly through frequent late-season queen supersedure. However, relative to targeted pupae, increased mortality particularly of heat-treated non-target brood stages (eggs and larvae) appeared to trigger compensatory colony-level responses, translating into temporarily decreased numbers of adult workers coupled with increased pollen foraging and overall lower honey harvests. Valuable insights into previously unrecognised side-effects of hyperthermia and mitigation thereof may ultimately permit successful routine applications of this chemical-free approach to combat the major threat to honeybees worldwide. [ABSTRACT FROM AUTHOR]

Simple Summary: Tuta absoluta (Meyrick) is an invasive insect pest that poses a threat to solanaceous crops. In recent years, the occurrences of extremely high temperature have been increasing due to climate change. Therefore, it is important to study the effect of high temperature on the growth and development of T. absoluta. In this study, when the eggs, pupae, and adults of T. absoluta were exposed to high-temperature stress, the pupal stage exhibited the highest sensitivity. This was demonstrated by a significant decrease in emergence rate, longevity, and fecundity. High-temperature stress during the egg stage resulted in a cascading effect, leading to a reduction in the adult longevity. This study provides a new insight for the integrated management of T. absoluta under high temperature. Insect life processes and reproductive behaviors are significantly affected by extremely high temperatures. This study focused on Tuta absoluta, which poses a severe threat to tomato cultivars. The effects of intense heat stress on the growth, development, oviposition, and longevity of T. absoluta were investigated. This investigation encompassed various developmental stages, including eggs, pupae, and adults. This study revealed that egg hatching and pupa emergence rates were significantly reduced at a temperature of 44 °C maintained for 6 h. The longevity of adults that emerged after the egg and pupal stages were exposed to 44 °C for 6 h was significantly reduced compared to the control. Notably, there was no significant variation in adult fecundity after egg-stage exposure to high temperatures. However, all treatments exhibited significantly reduced fecundity compared to the control after exposure to high temperatures during the pupal stage. Adult survival rates after exposure to 40 °C and 44 °C for 3 h were 74.29% and 22.40%, respectively, dramatically less than that of the control, which was 100%. However, no significant differences were noted in terms of longevity and egg production. These results offer a better understanding of the complex interactions between extreme temperatures and the life history traits of T. absoluta, thereby offering valuable insights for implementing management strategies to alleviate its impact on tomato crops in response to climate change. [ABSTRACT FROM AUTHOR]

One of the characteristics of the adult eclosion rhythm in Drosophila melanogaster is that adult emergence time differs greatly between the first and second eclosion days. The emergence time is in the middle of the light period on the first eclosion day, but immediately after light‐on on the second day. We hypothesized that incomplete entrainment of the endogenous pacemaker to the light–dark (LD) cycle is responsible for the daily variation. Due to the very short pupal period of this species, adult emergence may occur before complete synchronization of the pacemaker with the external cycle on the first eclosion day. Therefore, the peak time on the first eclosion day may differ significantly from that on the second day. To verify this hypothesis, using pupae that had pupariated within 24 h, the time difference between the first and second peaks was compared in LD 12:12 or constant darkness at five different temperatures from 30 to 15°C. In both light regimes, the time difference decreased with decreasing temperature and extended pupal duration. The interval between the eclosion peaks approached 24 h, supporting this hypothesis. These results can be interpreted by using a two‐oscillator model. [ABSTRACT FROM AUTHOR]

Morphological description of egg, larval chaetotaxy, pupa, adult genitalia of plume moth Sphenarches anysodactylus (Walker, 1864) (Pterophoridae: Lepidoptera) is presented. The mtCO1 of the Indian population done in this study is provided, revealing maximum similarity with six populations from Africa (97.55 to 98.37 %) and two populations from Australia (97.52 to 97.70) with differences in 10-15 nucleotide sequences. [ABSTRACT FROM AUTHOR]

IntroductionDengue is a public health challenge worldwide. Brazil registered about 70% of cases in Latin America in 2023; in 2024, the country is experiencing an unprecedented increase in the number of infected individuals. By May 2024, more than 4 million people were infected. Our goal was to: (1) determine the epidemiology of dengue cases and their spatiotemporal distribution and (2) carry out a survey of the storm drains and through a geospatial analysis to determine their possible correlation with cases of dengue in Presidente Prudente, São Paulo, Brazil.MethodsCases and information on the habitat of mosquito in the storm drain underground drainage system from 2020 to 2021 were obtained from public agencies. Larvae, pupae, and Ae. aegypti were identified according to species and described in taxonomic keys. Kernel density maps were constructed.ResultsFrom 1996 to 2023, the prevalence of cases peaked in 2016 and 2019, and in 2023 reached alarming levels, and the city was considered hyperendemic. In 2021, 2,609 cases were registered with 2 clusters of high density. Of 5,492 storm drains analyzed, 18.0% were found to have water, 9.0% had larvae or pupae of Aedes aegypti and 91.0% were classified as dirty or damaged. A direct correlation between the kernel layer of cases in 2021 with the kernel layer of storm drains containing water (r = 0.651) and larvae and pupae (r = 0.576) was found, suggesting that storm drains are risk factors and have an impact on the maintenance of dengue endemicity. The high number of damaged units found demonstrated the lack of storm drain management, compromising the urban drainage system and possibly contributing to dengue outbreaks.ConclusionPolicymakers may use these findings to improve existing dengue control strategies focusing on the control of storm drains and increase local and global perspectives on reducing dengue outbreaks.

Simple Summary: The concentration of larval black flies in well-defined aquatic habitats makes knowledge of the breeding sites critical for the efficient management of pest species. Black flies can achieve pest status either as generalists by developing in many types of streams and rivers or as specialists by developing in one or a few types of flowing water, such as large rivers. These two developmental strategies are evident in pest and vector species of the Simulium (Gomphostilbia) varicorne species group in Thailand. Simulium chumpornense is a habitat generalist, whereas S. khelangense is a habitat specialist, developing in the large Mekong River, where we discovered its immature stages. The first descriptions of the larva and pupa of S. khelangense, along with mitochondrial and nuclear genetic markers, allow accurate identification and comparisons with structurally similar species in the S. varicorne species group, thus aiding the ability to monitor the pest and vector status of black flies in Southeast Asia. Two species of black flies (Simuliidae) in Thailand, Simulium chumpornense Takaoka and Kuvangkadilok, 2000, and S. khelangense Takaoka, Srisuka & Saeung, 2022, are potent vectors of avian blood protozoa of the genera Leucocytozoon and Trypanosoma and are pests of domestic avian species. Although the adults are abundant throughout Thailand, information on their breeding habitats is limited, and the immature stages of S. khelangense are unknown. We collected the larvae and pupae of S. khelangense from the Mekong River, the first-ever record of Simuliidae from this large continental river. Mitochondrial cytochrome c oxidase I and internal transcribed spacer 2 were used to associate the larvae and pupae with known adults. Both genetic markers strongly supported their identity as S. khelangense. The larvae and pupa of S. khelangense are described. The pupal gill filaments, larval abdominal protuberances, and setae distinguish this species from other members of the S. varicorne species group. The immature stages of S. chumpornense inhabit a wide variety of flowing waters, from small streams (3 m wide) to enormous continental rivers (400 m wide); thus, S. chumpornense is a habitat generalist. In contrast, S. khelangense was found only in the large Mekong River and is, therefore, a habitat specialist. Both species can exploit their principal habitats and produce abundant adult populations. [ABSTRACT FROM AUTHOR]

Simple Summary: The silkworm has fascinated people all around the world for centuries due to numerous applications of by-products resulting from its metamorphosis. A decline has been noticed due to industrialization and some disease concerns; however, several reasons strengthen compelling arguments for keeping up this species. The most prevalent nutrients identified along silkworm rearing, from mulberry leaves up to larvae, pupae, and silk, are proteins. The biological value of protein is based on amino acids. The presence of other nutrients, especially fatty acids (n-3 family), vitamins, minerals, and polyphenols, along with the profile in amino acids, which are taken from mulberry leaves and metabolized by larvae and then transferred to pupae, strengthen the conviction according to which silkworm by-products have a great nutritional potential that is either comparable with fish meal or better than other aquaculture-specific feedstuffs. This paper aims to present deep knowledge about amino acids (classification, metabolism, occurrence, mechanisms, and potential functions) for silkworms and fish to further understand these species. The silkworm Bombyx mori, the second most varied group of insects, is a fascinating insect that belongs to the Lepidoptera species. We aimed to deepen our knowledge about the composition and significance of amino acids (AA) from the sericulture chain to fish. AAs are the most prevalent molecules throughout the growth process of silkworms. We described AAs classification, occurrence, metabolism, and functions. Online datasets revealed that the essential AAs (EAA) level in fish meal and silkworm pupae (SWP) is comparable. SWP have a high content of methionine and lysine, which are the principal limiting AAs in fish diets, indicating that SWP have nutritional potential to be added to fish diets. Additionally, an overview of the data analyzed displays that SWP have a higher protein efficiency ratio than fish meal, the classical protein-rich source (>1.19 times), and compared to soybean meal, the second-most preferred source of protein in aquaculture (>2.08 times), indicating that SWP can be considered effective for animal feeding. In this study, we provide an overview of the current knowledge concerning AAs, paying special emphasis to EAAs and explaining, to some extent, certain mechanisms and functions of these compounds, from mulberry leaves to larvae–pupae and fish diets. [ABSTRACT FROM AUTHOR]

The pine wood nematode (PWN) uses several Monochamus species as vehicles, through a temporary hitchhiking process known as phoresy, enabling it to access new host plant resources. Monochamus saltuarius acts as a new and major vector of the PWN in Northeastern China, showing lower PWN carrying capacity and a shorter transmission cycle compared to established vectors. The apparently altered symbiotic relationship offers an interesting area for researching the costs and adaptions involved in nematode–beetle, a specialized phoresy. We analyzed the response and fitness costs of M. saltuarius through physiological measurements and transcriptomics. The PWN exerted adverse repercussions on the growth and development of M. saltuarius. The PWN accelerated larval development into pupae, while beetle adults carrying the PWN exhibited an elevated abnormality rate and mortality, and reduced starvation resistance. During the pupal stage, the expression of growth-related genes, including ecdysone-inducible genes (E74EA), cuticle proteins, and chitin genes (CHTs), markedly increased. Meanwhile, the induced immune response, mainly by the IMD and Toll signaling pathways, could be a contributing factor to adult abnormality and mortality. Adult gonads and trachea exhibited enrichment in pathways related to fatty acid elongation, biosynthesis, and metabolism. FASN, ELOVL, and SCD possibly contributed to resistance against PWN. Our research indicated that phoretic interactions between vector beetles and PWN vary throughout the vector's lifespan, particularly before and after entry into the trachea. This study highlighted the fitness costs of immunity and metabolism on the vector beetle, indicating the adaptation mechanisms and evolutionary trade-offs to PWN. [ABSTRACT FROM AUTHOR]

Diapause is a peculiar sleep of insects in which the animal's motor activity completely ceases. During this period, insects become a good target for parasitoids, freely attacking them with their mobile ovipositors. We found that the parasitic wasp, Eupelmus messene (Hymenoptera, Eupelmidae, Cynipidae), stirs the internal contents of the diapausing host pupa of Aulacidea hieracii (Bouché, 1834) with its long and flexible ovipositor "making a shake" inside the pupa. However, the attacked pupae stay morphologically indistinguishable from healthy diapausing ones for several months. Using non-invasive Raman spectroscopy (RS), we, for the first time, studied the molecular composition of live diapausing and parasitized A. hieracii pupae. As a result of the study, we discovered alterations in the content and distribution of carotenoids, lipids, and proteins in the attacked pupae compared to the live intact diapausing pupae. [ABSTRACT FROM AUTHOR]

The system of gulcosinoalte sulfatase–sulfatase‐modifying factors (Sumfs) in herbivorous insects fulfils an essential function in defending host plant glucosinolates (GLSs)—the compounds of cruciferous plants—by swiftly desulfurising them. Previously, eight HaSulfs and one HaSumf1 were identified in Helicoverpa armigera. GLSs have a significant impact on the growth and development of H. armigera larvae. They can induce the expression of HaSulf in the midgut of H. armigera at both gene and protein levels. In the present study, we utilised RNAi technology to silence the HaSumf1 gene and explored its potential impact on the growth, development, and adaptation of GLSs. The sequence analysis has indicated that the HaSumf1 gene in H. armigera shares a high degree of sequence identity with other insect genes and is expressed at its highest level in the fifth‐instar phase. Additionally, the qRT‐PCR results reveal that the expression of HaSumf1 was upregulated by 1.26 and 0.07 times in third‐instar and 1.65 and 0.19 times in fifth‐instar when fed with 0.004% and 0.04% GLS, respectively. The effectiveness of dsSumf1 silencing was observed in both third‐ and fifth‐instar stages, with silencing rates of 65.5%–88.9% and 61.3%–74.6%, respectively (p < 0.05). The findings of the bioassay indicate that H. armigera's body weight at fifth‐instar significantly increased after treatment with 0.004% GLS and 0.04% GLS, while the larval survival rate, pupation rate, and pupa weight decreased. Notably, when feeding dsRNA‐encapsulated carbon quantum dots (CQD‐dsRNA), this trend was significantly strengthened (p < 0.05). Taken together, these findings provide evidence supporting HaSumf1's conservation involvement in the adaptation of host plant GLSs during lepidopteran insect evolution. Therefore, it plays a critical role in the growth and development of H. armigera, offering molecular targets for developing novel pesticides and targeted control measures against this vital pest. [ABSTRACT FROM AUTHOR]

Xylotrechus rusticus is one of the major forest trunk borers in China, causing a significant threat to afforestation and forestry engineering construction. Controlling X. rusticus is challenging because of its hidden life habit. The use of natural enemies against X. rusticus is an important strategy for controlling Cerambycidae pests. This is the first study to investigate the effectiveness of Dastarcus helophoroides as a biological control against X. rusticus under indoor and outdoor environments. The expression patterns of CYP6BK3 and CYP6BQ13 genes in various tissues and at different development stages were analyzed to clarify their role in the development of D. helophoroides. The results showed that the parasitism rate of D. helophoroides was the higher when their eggs were released on the larvae of X. rusticus L. indoors. Furthermore, the parasitism rate of D. helophoroides was the highest (86.67%) when their eggs were released on the larvae of X. rusticus in the ratio of 1:100 (host: D. helophoroides) under artificial wood segment simulation. The average parasitism rates of D. helophoroides eggs and adults on the exposed wood segment of the Cerambycidae were 81.83% and 83.29%, respectively. Indoor release of Scleroderma guani increased their parasitic rate on the aged larvae and pupae of X. rusticus. qPCR analysis of CYP450 gene expression showed that CYP6BK3 and CYP6BQ13 genes were expressed at all development stages of D. helophoroides; however, their expression levels were higher in adults than in the larvae. Notably, CYP6BK3 expression was the highest in the adipose tissue, whereas CYP6BQ13 was the highest in the hindgut tissue. [ABSTRACT FROM AUTHOR]