Courtship and mating behavior of Endoclita signifer (Hepialidae: Lepidoptera) and the male sex pheromones in hairbrushes.

Endoclita signifer Walker is the most destructive wood-boring pest of Eucalyptus in China, causing significant economic and ecological damage. As an insect of the primitive Lepidoptera family Hepialidae, E. signifer fly and mat for only 10-20 min at dusk. The courtship and mating behavior of E. signifer adults and whether male moths release sex pheromones are still unknown, especially since transitory flight survival strategies in primitive moths differ from advanced moths like noctuids. In this study, we first observed the courtship and mating behavior of E. signifer by considering the effects of space and then analyzed extracts of male hairbrushes using gas chromatography-electroantennogram detection. Our results indicated that during the courtship period, flying males form courtship fields, lekking, and chase flying females before mating with them; E. signifer were more successful in mating in larger spaces (Length × Width × Height = 9.6 × 7 × 4 m); 5 compounds in the hairbrushes of the male moths which elicited antennal responses of 2 sexes, despite at high concentrations. Combined with it, indicating that communication between male and female may rely on male sex pheromones. These findings can serve as a basis for studying the mechanisms of sex communication in E. signifer and developing sex pheromone-based trapping techniques.

Identification and Validation of Reference Genes for Expression Analysis Using RT-qPCR in Leptocybe invasa Fisher and La Salle (Hymenoptera: Eulophidae).

Leptocybe invasa (Hymenoptera: Eulophidae) is a globally intrusive pest. Despite extensive research into the physiological responses of this pest, our understanding of the molecular mechanisms still needs to be improved. We want to accurately investigate the expression of L. invasa's target genes, so it is imperative to select fitting reference genes. In this study, eight housekeeping genes' stability (RPS30, ACTR, 18S rRNA, ACT, RPL18, GAPDH, 28S rRNA, and TUB) was tested under five different experimental conditions, including male or female adults, somites (head, thorax, and abdomen), temperatures (0 °C, 25 °C, and 40 °C), diets (starvation, clear water, 10% honey water, Eucalyptus sap), and pesticides (acetone was used as a control, imidacloprid, monosultap). Gene stability was calculated using RefFinder, which integrates four algorithms (the ∆Ct method, geNorm, NormFinder, and BestKeeper). The findings implied that ACT and ACTR were the most accurate when comparing sexes. For analyzing different somites, 28S rRNA and RPL18 were ideal; the 28S rRNA and RRS30 were perfect for analyzing at different temperatures. The combination of ACT and GAPDH helped to analyze gene expression in different diets, and GAPDH and 28S rRNA were suitable for various pesticide conditions. Overall, this research offers a complete list of reference genes from L. invasa for precise analysis of target gene expression, which can improve the trustworthiness of RT-qPCR and lay the foundation for further investigations into the gene function of this pest.

Quick shift in volatile attraction between the third and fifth instar larvae of Endoclita signifier.

BACKGROUND: Endoclita signifer is a polyphagous lepidopteran species of eucalyptus that selects its hosts in the third-instar larval period. To understand how it adapts to its host during development, we studied the olfactory responses of late-stage (fifth)-instar larvae to the dynamic chemical environment they encounter. RESULTS: Thirty-two volatiles from eucalyptus trunk and soil were identified, among which 14 showed electroantennal activity and five were identified as new. Further behavioral bioassay showed that both ß-pinene and the imitation ratio of six key volatile in eucalyptus trunk were attractive to the fifth-instar larvae, but both eliminated and increased ß-pinene in the mixture decreased the choice ratio and showed no attraction. Although E. signifer larvae shifted their attraction from o-cymene at the third-instar stage to ß-pinene at the fifth-instar stage in a single volatile, the appropriate ratio of the main compounds in eucalyptus trunk volatile is the key to the behavior choice of fifth-instar larvae. CONCLUSION: The switch in olfactory attraction to different compounds between fifth- and third-instar larvae indicated an olfactory plasticity between third- and fifth-instar larvae. And the particular blend mediated the highly specialized communication interactions specificity between fifth-instar larvae and eucalyptus trunk volatile indicated the specialized host adaptation to fifth-instar larvae. This enhances understanding of how the primitive lepidopteran E. signifier, as a native pest, adapts to introduced eucalyptus. Moreover, this study provides knowledge for the screening and development of target volatiles for trapping and managing E. signifer larvae. © 2022 Society of Chemical Industry.

Antibiotics use and risk of colorectal neoplasia: An updated meta-analysis.

PURPOSE: Accumulating evidence indicate that antibiotic use could induce microbiome dysbiosis, which was a critical driver to the onset and progression of colorectal cancer (CRC). But the relationship between antibiotics use and CRC was still disputed. Hence, we conducted this systematic review and meta-analysis to appraise and synthesize the present available evidence to clarify the association. METHODS: PubMed, Embase, Web of Science, and Cochrane Library were systematically searched for relevant observational studies from inception to June 5, 2020. Pooled odds ratios (ORs) with 95% confidence intervals (CIs) were estimated to explore the association between antibiotics use and CRC using random-effects model. Subgroup analyses, sensitive analyses, and publication bias were conducted to assess the robust reliability of pooled results. RESULTS: A total of 15 observational studies containing 5,164,138 patients were included in this meta-analysis. The pooled analysis indicated that the total antibiotic use was correlated with increased risk of CRC (OR, 1.11; 95% CI, 1.05-1.18). The subgroup analyses suggested that antibiotic use significantly elevated risk of colon cancer, but not rectal cancer. Furthermore, we found that penicillin, cephalosporin, anti-anaerobic, and anti-aerobic antibiotics increased the risk of CRC, in particular metronidazole but no significant associations were identified in macrolide, tetracycline, sulfonamides, nitrofurans, and quinolone use. The results of sensitive analyses and publication bias indicated the conclusions were robust. CONCLUSION: The findings showed that antibiotics use may be associated with the onset of CRC. Policy-makers and clinicians should adequately assess possible benefits and harms of antibiotics use, especially in some high-risk populations. Also, for high-risk patients with previous antibiotics use, it was suggested to perform early colonoscopy screening to find or even eliminate early-stage CRC.

EsigGOBP1: The Key Protein Binding Alpha-Phellandrene in Endoclita signifer Larvae.

Endoclita signifer larvae show olfactory recognition towards volatiles of eucalyptus trunks and humus soils. Further, EsigGOBP1 was identified through larval head transcriptome and speculated as the main odorant-binding proteins in E. signifer larvae. In this study, the highest expression of EsigGOBP1 was only expressed in the heads of 3rd instar larvae of E. signifer, compared with the thorax and abdomen; this was consistent with the phenomenon of habitat transfer of 3rd instar larvae, indicating that EsigGOBP1 was a key OBP gene in E. signifer larvae. Results of fluorescence competition binding assays (FCBA) showed that EsigGOBP1 had high binding affinities to eight GC-EAD active ligands. Furthermore, screening of key active odorants for EsigGOBP1 and molecular docking analysis, indicated that EsigGOBP1 showed high binding activity to alpha-phellandrene in 3rd instar larvae of E. signifer. Conformational analysis of the EsigGOBP1-alpha-phellandrene complex, showed that MET49 and GLU38 were the key sites involved in binding. These results demonstrated that EsigGOBP1 is a key odorant-binding protein in E. signifer larvae, which recognizes and transports eight key volatiles from eucalyptus trunk, especially the main eucalyptus trunks volatile, alpha-phellandrene. Taken together, our results showed that EsigGOBP1 is involved in host selection of E. signifer larvae, which would aid in developing EsigGOBP1 as molecular targets for controlling pests at the larval stage.