Neuropeptides play vital roles in energy homeostasis in insects and other animals. Although the importance of the regulatory network of neuropeptides in feeding and metabolism has been illuminated, a complete understanding of the mechanisms has not been addressed as many factors are involved in the regulation. CCHamide-2 is a newly identified brain-gut neuropeptide that regulates feeding behavior in several insect species including Drosophila melanogaster. However, little is known about the mechanisms controlling the feeding-related behavior and metabolic functions modulated by CCHamide-2 in other insects. In this study, we addressed the functions of CCHamide-2 in the two-spotted cricket, Gryllus bimaculatus, which was used as the experimental material to research the mechanisms of feeding and metabolism in this omnivorous insect species. Knockdown crickets by RNA interference against GbCCHamide-2R increased the amount of food intake, while injection of chemically synthetic GbCCHamide-2 peptide reduced the amount of food intake. Further, knockdown and peptide injection experiments revealed that GbCCHamide-2 signaling increased the concentrations of circulating lipids and carbohydrates, and the carbohydrate-rich diet increased the transcript levels of GbCCHa-2R. Moreover, GbCCHa-2 injection decreased the transcript level of Gbilp. By contrast, GbCCHamide-2 signaling did not affect nymphal growth or the transcript level of GbAKH, as well as feeding preferences. Taken together, CCHamide-2 signaling in G. bimaculatus regulates food intake associated with alterations in lipid and carbohydrate levels in hemolymph.

Antibiotic resistance genes (ARGs) have become recognized contaminants and pose a high public health risk. The animal gut microbiota is a reservoir of ARGs, but the knowledge of the origin and dissemination of ARGs remains unclear. In this study, we provide a comprehensive profile of ARGs and mobile genetic elements in the gut microbiota from 30 bovines to study the impact of modern antibiotics on resistance. A total of 42 ARG types were detected by annotating the metagenomic sequencing data from Comprehensive Antibiotic Resistance Database (CARD). We found that the diversity and abundance of ARGs in individual yaks were significantly lower than those in dairy and beef cattle ( p < 0.0001). The results of heat map and single nucleotide polymorphism clustering suggest that ARGs from dairy and beef cattle are more similar, whereas those from yaks cluster separately. The long-term use of antibiotics may contribute to this difference, suggesting that antibiotic consumption is the main cause of ARG prevalence. Furthermore, abundant insertions were also found in this study, signifying a strong potential for horizontal transfer of ARGs among microbes, especially pathogens.

The brown planthopper Nilaparvata lugens is a typical monophagous insect herbivore that feeds exclusively on rice sap. This insect pest causes serious damage to rice crops throughout East Asian countries. Chemical control remains the first choice for managing N. lugens populations; however, the use of insecticides has given rise to planthopper resurgence and additional environmental risks. Nilaparvata lugens is a model insect of Hemiptera because its whole genome sequence has been elucidated and is susceptible to RNA interference. In this study, our findings revealed that a superoxide-generating gene, NADPH oxidase 5 (Nox5), is essential for molting and oviposition in a Hemipteran insect Nilaparvata lugens. Knockdown of Nox5 transcript levels by RNA interference in 2nd–5th-instar nymphs results in significantly lethal deficits in the molting transitions from nymph–nymph and nymph–adult. Nox5 knockdown leads to a reduction of hydrogen peroxide in female ovaries and failure of oviposition from the insect ovipositor into the rice leaf sheath. Here, we provide in vivo evidence demonstrating that Nox5 is a key enzyme for regulating molting and oviposition in this insect species.

Genome annotation is critically important data that can support research. Draft genome annotations cover representative genes; however, they often do not include genes that are expressed only in limited tissues and stages, or genes with low expression levels. Neuropeptides are responsible for regulation of various physiological and biological processes. A recent study disclosed the genome draft of the two-spotted cricket Gryllus bimaculatus, which was utilized to understand the intriguing physiology and biology of crickets. Thus far, only two of the nine reported neuropeptides in G. bimaculatus were annotated in the draft genome. Even though de novo assembly using transcriptomic analyses can comprehensively identify neuropeptides, this method does not follow those annotations on the genome locus. In this study, we performed the annotations based on the reference mapping, de novo transcriptome assembly, and manual curation. Consequently, we identified 41 neuropeptides out of 43 neuropeptides, which were reported in the insects. Further, 32 of the identified neuropeptides on the genomic loci in G. bimaculatus were annotated. The present annotation methods can be applicable for the neuropeptide annotation of other insects. Furthermore, the methods will help to generate useful infrastructures for studies relevant to neuropeptides.

After mating, some insects drastically reduce mating activity, termed post-mating refractoriness, due to physiological restrictions. Male crickets, Gryllus bimaculatus, characteristically exhibit a 1-hour post-mating refractory stage, which is controlled by terminal abdominal ganglion. The molecular mechanisms underlying the male-specific precisely timed refractory stage remain unclear. Here, we show that among 28 neuropeptide precursors expressed in the terminal abdominal ganglion, myosuppressin, allatotropin, and sNPF exhibited male-specific expression based on RT-qPCR and in situ hybridization. RNA interference experiments showed that only the knockdown of allatotropin and sNPF extended the refractoriness duration. Furthermore, allatotropin and sNPF knockdown influenced the male reproductive system by inhibiting seminal fluid secretion from the male accessory gland and decreasing spermatozoon storage in seminal vesicles, respectively. Knockdown of allatotropin and sNPF receptors caused similar phenotypes to their ligands. In conclusion, this study demonstrated the regulation of post-mating refractoriness and reproductive system by Allatotropin and sNPF signaling pathways in male crickets.

Ion transport peptide (ITP) and ITP-like (ITPLs) are pleiotropic bioactive peptides in insects. Although the contribution of these peptides to ecdysis has been studied, the precise regulatory mechanisms remain poorly understood. Here, we characterized the functions of itp and itpl variants in the two-spotted cricket, Gryllus bimaculatus. Reverse transcription-quantitative PCR and whole-mount in situ hybridization revealed that itp was expressed in the brain and terminal abdominal ganglion, whereas itpl variants were expressed in all ganglia of the central nervous system. Simultaneous knockdown of itp and itpls disrupted ecdysis behavior and water transport from the gut into the hemolymph during molting. Nevertheless, knockdown of itpls without influencing itp expression did not significantly affect ecdysis behavior but caused a reduction in hemolymph mass. Although water transport into the hemolymph is considered necessary for the swelling required to split the old cuticle layers during molting, a rescue experiment by injection of water or cricket Ringer's solution into the hemolymph of knockdown crickets did not recover the normal phenotype. Therefore, we propose that ITP/ITPL control ecdysis behavior probably not by regulating water transport from the gut into the hemolymph in crickets.