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Copidosoma floridanum is a cosmopolitan species and an egg-larval parasitoid of the Plusiine moth. C. floridanum has a unique development mode called polyembryony, in which over two thousand genetically identical embryos are produced from a single egg. Some embryos develop into sterile soldier larvae precociously, and their emergence period and aggressive behavior differ between the US and Japanese C. floridanum strains. Genome sequencing expects to contribute to our understanding of the molecular bases underlying the progression of polyembryony. However, only the genome sequence of the US strain generated by the short-read assembly has been reported. In the present study, we determined the genome sequence of the Japanese strain using Pacific Biosciences high-fidelity reads and generating a highly contiguous assembly (552.7 Mb, N50: 17.9 Mb). Gene prediction and annotation identified 13,886 transcripts derived from 10,786 gene models. We searched the genomic differences between US and Japanese strains. Among gene models predicted in this study, 100 gene loci in the Japanese strain had extremely different gene structures from those in the US strain. This was accomplished through functional annotation (GGSEARCH) and long-read sequencing. Genomic differences between strains were also reflected in amino acid sequences of vasa that play a central role in caste determination in this species. The genome assemblies constructed in this study will facilitate the genomic comparisons between Japanese and US strains, leading to our understanding of detailed genomic regions responsible for the ecological and physiological characteristics of C. floridanum. [ABSTRACT FROM AUTHOR]

(1) The global population is projected to reach a staggering 9.8 billion people by the year 2050, leading to major concerns about food security. The necessity to increase livestock production is inevitable. The black soldier fly (BSF) is known for its ability to consume a wide range of organic waste, and BSF larvae have already been used as a partial substitute for fishmeal. In contrast, the use of antibiotics in livestock feed for growth promotion and prophylaxis poses a severe threat to global health owing to antimicrobial resistance. Insect antimicrobial peptides (AMPs) have shown the potential to rapidly disrupt target bacterial membranes, making bacterial resistance to AMPs a less likely concern. (2) In this study, we explored various methods for stimulating AMP synthesis in BSF larvae and found that thermal injury effectively induced the production of various AMP types. Additionally, we investigated the activation of innate immune response pathways that lead to AMP production following thermal injury. (3) Interestingly, thermal injury treatment, although not involving bacteria, exhibited a similar response to that observed following Gram-positive bacterial infection in eliciting the expression of AMP genes. (4) Our findings offer support for the industrial use of BSF to enhance livestock production and promote environmental health. [ABSTRACT FROM AUTHOR]

Simple Summary: The functions of all genes encoded in the genome should be studied for genome editing. Genome editing technology can speed up insect research for the functional analysis of genes. Our knowledge about the functional information of genes is still incomplete currently, while the genome sequencing of an organism can be completed. The functional information has been annotated based solely on the information that has been obtained from the results of previous biological research. However, this information will be important in determining the target genes for genome editing. In particular, it is very important that this information is in machine-readable form because computer programs mainly parse this information for the understanding of biological systems. In this paper, we describe a workflow-based method for annotating gene functions in insects that makes use of transcribed sequence information as well as reference genome and protein sequence databases. Using the developed workflow, we annotated the functional information of the Japanese stick insect and silkworm, including gene expression as well as sequence analysis. The functional annotation information obtained by the workflow will greatly expand the possibilities of entomological research using genome editing. Next-generation sequencing has revolutionized entomological study, rendering it possible to analyze the genomes and transcriptomes of non-model insects. However, use of this technology is often limited to obtaining the nucleotide sequences of target or related genes, with many of the acquired sequences remaining unused because other available sequences are not sufficiently annotated. To address this issue, we have developed a functional annotation workflow for transcriptome-sequenced insects to determine transcript descriptions, which represents a significant improvement over the previous method (functional annotation pipeline for insects). The developed workflow attempts to annotate not only the protein sequences obtained from transcriptome analysis but also the ncRNA sequences obtained simultaneously. In addition, the workflow integrates the expression-level information obtained from transcriptome sequencing for application as functional annotation information. Using the workflow, functional annotation was performed on the sequences obtained from transcriptome sequencing of the stick insect (Entoria okinawaensis) and silkworm (Bombyx mori), yielding richer functional annotation information than that obtained in our previous study. The improved workflow allows the more comprehensive exploitation of transcriptome data and is applicable to other insects because the workflow has been openly developed on GitHub. [ABSTRACT FROM AUTHOR]

Temperature is one of the most influential factors for animals. The acclimation (rearing) and challenge temperatures are often more important than the given temperature per se. These effects on physiological responses have been known, but not well understood on immune responses. Here, we investigated the integrated effects of rearing and challenge temperatures on haemocyte populations in larvae of a plusiine moth, Chrysodeixis eriosoma. We hypothesize that the haemocyte concentration is decreased (increased) at higher (lower) temperatures from rearing temperatures and that the proportions of haemocyte types exhibit directional changes at higher (lower) temperatures to compensate for immune reactions. We expect that increasing (decreasing) the challenge temperature from the rearing temperature enhances (reduces) phagocytic activity. We found that higher temperatures slightly decreased the haemocyte concentration. We detected small changes in the proportions of haemocyte types among rearing temperatures, but the changes were non‐directional and most of them were statistically insignificant. We also found the integrated effects only with increases in the challenge temperatures, which resulted in increased phagocytosis, whereas no apparent reactions were detected with decreases in the challenge temperatures. Our results show that the haemocyte concentration is significantly affected by the rearing temperature, which implies that haematopoiesis depends on the ambient temperature. We discuss some adaptive and non‐adaptive components for the positive integrated effects of increases in the challenge temperatures. We also discussed the obtained non‐responsiveness in the integrated effects with decreases in the challenge temperatures. [ABSTRACT FROM AUTHOR]

Copidosoma floridanum is a polyembryonic, caste-forming, wasp species. The ratio of investment in different castes changes with environmental stressors (e.g. multi-parasitism with competitors). The vasa gene was first identified in Drosophila melanogaster as a germ-cell-determining factor, and C. floridanum vasa (Cf-vas) gene positive cells have been known to develop into reproductive larvae. Cf-vas seems to control the ratio of investment in C. floridanum larval castes. In this study, we identified environmental factors that control Cf-vas mRNA expression in Japanese C. floridanum by examining Cf-vas mRNA expression under competitor (Meteorus pulchricornis) venom stress; we treated the male and female morulae with M. pulchricornis venom. We also assessed the effects of multi-parasitism of Japanese C. floridanum with M. pulchricornis and found an increasing number of female soldier larvae. The results showed that several amino acid sequences differ between the Japanese and US Cf-vas. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) showed that Japanese Cf-vas mRNA is expressed in both male and female larvae and pupae, but mRNA expression decreases in adults. Cf-vas mRNA expression significantly decreased, while C. floridanum dronc (Cf-dronc) mRNA expression increased, in female morulae after M. pulchricornis venom treatment at 20 h and 0 h of the culture period, respectively. Females and males showed different Cf-vas or Cf-dronc mRNA expression after M. pulchricornis venom treatment. Therefore, M. pulchricornis venom could affect the ratio of investment in different female castes of Japanese C. floridanum by decreasing Cf-vas mRNA expression via apoptosis. Image 1 • Japanese Cf-vas and US Cf-vas amino acid sequences were different. • Co-parasitism with a competitor significantly increased the number of female, but not male soldier larvae. • Japanese Cf-vas mRNA expression decreased, while Cf-dronc mRNA expression increased, in female morulae after venom treatment. • M. pulchricornis venom increases the number of female soldier larvae by decreasing Japanese Cf-vas mRNA expression. [ABSTRACT FROM AUTHOR]

Larval castes are well known in both sexes of polyembryonic parasitic wasps, in which sterile soldier larvae are clonally produced from a single egg and thousands of reproductive larvae develop into adults. The proportion of soldier larvae in Copidosoma floridanum is first determined genetically in both sexes but subsequently increases in females as a result of environmental disturbances, such as the presence of competitive parasitoids. Because temperature is the most influential environmental factor affecting ectotherms, we hypothesize that an increase in ambient temperature also affects the production of female soldier larvae. Conversely, we hypothesize that there would be no effect of temperature on the production of male soldier larvae. In the present study, we show that heat stress experienced by C. floridanum at an early stage in host development increases the number of female soldier larvae produced during the last instar stage of the host. We also show that rearing at extreme (high and low) temperatures induces an increase in the number of female soldiers produced. The effect of heat stress and ambient temperature on soldier production is found only in females, as is the response to heterospecific competitors. By contrast, no responses are detected in males for either heat stress or ambient temperature. The adaptive effect of these phenomena is not known, although the results of the present study imply the existence of a common physiological cascade that causes an increase in the number of soldier larvae in females. [ABSTRACT FROM AUTHOR]

Heavy sea-water spray icing is a major problem in the maintenance of lighthouses situated in northern har-bors facing the Sea of Japan. Moreover, low ice adhesion materials or anti-icing coatings are ineffective over long periods. In this study, we have investigated the characteristics of saline ice adhesion on hydrophilic and hydrophobic pliable polymer sheets that can be used to envelop and thereby protect small lighthouses. We conducted experiments in cold room laboratories, and further, we verified the deicing conditions. Ice adhe-sion strength tests using fresh water indicated that a highly hydrophobic surface exhibited lower adhesion strength than other test materials. The results of saline ice adhesion tests at --20°C indicated that the ad-hesion strength decreases very rapidly as the salinity increases for all materials; this trend was particularly remarkable in case of superhydrophilic materials. The water spray icing tests were verified by experiments conducted using an ice model basin. Wet growth of ice occurred on all test specimens including the highly hydrophobic sheets. Furthermore, the initial icing depended significantly on the hydrophobic or hydrophilic surface properties. The icing on the hydrophobic surface was characterized by the formation of vertical streaks of ice; irregular projections could be observed on the ice surface. On the other hand, a thin fluid flow was formed on superhydrophilic surfaces. Exfoliation of pieces of the sheet ice was often observed during the initial growth. Deicing could be easily performed for both pliable polymer sheets. A combination of low ice adhesion property and rapid distortion was effective for the deicing of sea-water spray icing. [ABSTRACT FROM AUTHOR]

The DJ-1 gene is highly conserved across a wide variety of organisms and it plays a role in anti-oxidative stress mechanisms in cells. The red flour beetle, Tribolium castaneum, is widely used as a model insect species because it is easy to evaluate gene function in this species using RNA interference (RNAi). The T. castaneum DJ-1 (TcDJ-1) sequence is annotated in the T. castaneum genome database; however, the function and characteristics of the TcDJ-1 gene have not been elucidated. Here, we investigated the cDNA sequence of TcDJ-1 and partially characterized its function. First, we examined the TcDJ-1 amino acid sequence and found that it was highly conserved with sequences from other species. TcDJ-1 mRNA expression was higher in the early pupal and adult developmental stages. We evaluated oxidant tolerance in TcDJ-1 knockdown adults using paraquat and found that adults with TcDJ-1 knockdown exhibited increased sensitivity to paraquat. Our findings show that TcDJ-1 has an antioxidant function, as observed for DJ-1 from other insects. Therefore, these results suggest that TcDJ-1 protects against oxidative stress during metamorphosis. [ABSTRACT FROM AUTHOR]