The mandibles are among the most important appendages of insects\' mouthparts. Their morpho-functional organization is correlated with the variation in dietary preferences. In this study, we investigated the ultrastructural organization and metal composition of the mandibles of two ladybird species with different dietary habits: Harmonia axyridis (an entomophagous species) and Subcoccinella vigintiquatuorpunctata (a phytophagous species). The ultrastructural organization was studied using Scanning and Transmission Electron Microscopy, whereas the metal composition was investigated using Energy-Dispersive X-ray spectroscopy (EDX). Significant differences were observed in the general organization and metal enrichment pattern between the two species. The mandibles of H. axyridis are large and present a molar part with two teeth, with the apical one showing a bifid apex. In contrast, S. vigintiquatuorpunctata exhibited a molar region with several teeth on its apical part. The study revealed significant differences in metal content between the teeth and the prostheca of H. axyridis. Mn was the most abundant element in teeth, whereas Cl was more abundant in the prostheca. In the case of S. vigintiquatuorpunctata, Si was the most abundant element in the prostheca, while Mn was more present in the teeth. A comparison between the two species revealed that both teeth and prostheca showed significant variation in the elemental composition. These findings underscore the role of dietary preferences in shaping the structural and metal composition variations in the mandibles of these two ladybird species.

With the development of biological control methods, the predatory ladybird beetle Harmonia axyridis Pallas (Coleoptera: Coccinellidae) has been widely used for pest control in agricultural production. Appropriate shelf-life management strategies could synchronize H. axyridis production with pest outbreaks, finally improving the effectiveness of biological control. Herein, we preliminarily explored whether an artificial diet could optimize the shelf-life management of H. axyridis. We compared the survival rate, nutrition accumulation, reproductive development, juvenile hormone (JH) related-gene expression levels, and stress resistance gene expression levels between aphid-fed and artificial diet-fed H. axyridis females. The results revealed that H. axyridis females maintained a high survival rate after being fed an artificial diet for 60 days, whereas the survival rate of aphid-fed females decreased. Continuous feeding of the artificial diet caused H. axyridis females to enter a diapause-like state, which was characterized by low JH levels, high triglycerides and trehalose accumulation, ovarian development inhibition, decreased Vgs expression levels, and increased stress resistance gene expression levels. This diapause-like state could be promptly recovered upon transferring to an aphid diet. These results indicate that the artificial diet could manipulate the reproductive development status of H. axyridis and lay the foundation for its shelf-life management.

Bacteria have a profound influence on life history and reproduction of numerous insects, while the associations between hosts and bacteria are substantially influenced by environmental pressures. Cold storage is crucial for extending the shelf life of insects used as tools for biological control, but mostly causes detrimental effects. In this study, we observed a great decrease in egg hatch rate of cold-stored Harmonia axyridis during the later oviposition periods. Furthermore, most eggs produced by their F1 offspring exhibited complete loss of hatchability. We hypothesized that long-term exposure to cold may greatly alter the bacterial community within the reproductive tracts of H. axyridis, which may be an important factor contributing to the loss of egg viability. Through sequencing of the 16S rRNA gene, we discovered considerable changes in the bacterial structure within the reproductive tracts of female cold-stored beetles (LCS_F) compared to non-stored beetles (Control_F), with a notable increase in unclassified_f_Enterobacteriaceae in LCS_F. Furthermore, in accordance with the change of egg hatchability, we observed a slight variation in the microbial community of eggs produced by cold-stored beetles in early (Egg_E) and later (Egg_L) oviposition periods as well as in eggs produced by their F1 offspring (Egg_F1). Functional predictions of the microbial communities revealed a significant decrease in the relative abundance of substance dependence pathway in LCS_F. Moreover, this pathway exhibited relatively lower abundance levels in both Egg_L and Egg_F1 compared to Egg_E. These findings validate that long-term cold storage can greatly modify the bacterial composition within H. axyridis, thereby expanding our understanding of the intricate bacteria-insect host interactions.

Harmonia axyridis (H. axyridis) is the natural enemy of many aphid species. Traditional manual release of H. axyridis adults requires substantial manpower, and release efficiency is low. Automatic mechanical devices can improve the efficiency of delivery. Based on H. axyridis adults\' morphological size, a prototype release system for H. axyridis was designed, which considered the adhesion characteristics of H. axyridis adults. According to the measured physical characteristics of H. axyridis adults, the structural parameters of the mechanical system for the release of the H. axyridis adults were determined. The relationship of the quantity of release, the impeller rotating speed, and the time for the release of H. axyridis adults were constructed. The mechanism can quantitatively adjust the number of H. axyridis adults to meet a certain H. axyridis-aphids ratio. Combining the image processing technology with the camera function of a mobile phone, the maximum cross-sectional area method was used to count the H. axyridis adults in the designated area. Results showed that the impeller rotating speed had a significant effect on the survival rate of the H. axyridis adults. When the airflow velocities were 29.5 m/s and 38.3 m/s, the survival rates of the H. axyridis adults were 93.8% and 94.5% at 4.2 rpm. The adhesion rate of the H. axyridis adults was 2.5%-4.6%. This work will provide technical support for the research of biological control.

Plant volatile organic compounds (VOCs) are the key distress signals involved in tritrophic interactions, by which plants recruit predators to protect themselves from herbivores. However, the effect of nitrogen fertilization on VOCs that mediate tritrophic interactions remains largely unidentified. In this study, a maize (Zea mays)-aphid (Rhopalosiphum padi)-ladybird (Harmonia axyridis) tritrophic interaction model was constructed under high-nitrogen (HN) and low-nitrogen (LN) regimens. H. axyridis had a stronger tendency to be attracted by aphid-infested maize under HN conditions. Then, volatiles were collected and identified from maize leaves on which aphids had fed. All of the HN-induced volatiles (HNIVs) elicited an electroantennogram (EAG) response from H. axyridis. Of these HNIVs, 1-nonene was attractive to H. axyridis under simulated natural volatilization. Furthermore, our regression showed that the release of 1-nonene was positively correlated with H. axyridis visitation rates. Supplying 1-nonene to maize on which aphids had fed under LN enhanced attractiveness to H. axyridis. These results supported the conclusion that 1-nonene was the active compound that mediated the response to nitrogen in the tritrophic interaction. In addition, the 1-nonene synthesis pathway was hypothesized, and we found that the release of 1-nonene might be related to the presence of salicylic acid (SA) and abscisic acid (ABA). This research contributes to the development of novel environmentally friendly strategies to optimize nitrogen fertilizer application and to improve pest control in maize crops.

The essential oil of Pelargonium graveolens (PGEO) is identified in the literature as a rich source of bioactive compounds with a high level of biological activity. This study aimed to examine the chemical profile of PGEO as well as its antioxidant, antibacterial, antibiofilm, and insecticidal properties. Its chemical composition was analyzed using gas chromatography-mass spectrometry (GC-MS), achieving comprehensive identification of 99.2% of volatile compounds. The predominant identified compounds were β-citronellol (29.7%) and geraniol (14.6%). PGEO\'s antioxidant potential was determined by means of DPPH radical and ABTS radical cation neutralization. The results indicate a higher capacity of PGEO to neutralize the ABTS radical cation, with an IC50 value of 0.26 ± 0.02 mg/mL. Two techniques were used to assess antimicrobial activity: minimum inhibitory concentration (MIC) and disk diffusion. Antimicrobial evaluation using the disk diffusion method revealed that Salmonella enterica (14.33 ± 0.58 mm), which forms biofilms, and Priestia megaterium (14.67 ± 0.58 mm) were most susceptible to exposure to PGEO. The MIC assay demonstrated the highest performance of this EO against biofilm-forming S. enterica (MIC 50 0.57 ± 0.006; MIC 90 0.169 ± 0.08 mg/mL). In contrast to contact application, the assessment of the in situ vapor phase antibacterial activity of PGEO revealed significantly more potent effects. An analysis of antibiofilm activity using MALDI-TOF MS demonstrated PGEO\'s capacity to disrupt the biofilm homeostasis of S. enterica growing on plastic and stainless steel. Additionally, insecticidal evaluations indicated that treatment with PGEO at doses of 100% and 50% resulted in the complete mortality of all Harmonia axyridis individuals.

BACKGROUND: Harmonia axyridis is an effective natural enemy insect to a variety of phloem-sucking pests and Lepidopteran larvae, such as aphids, scabies, and phylloxera, while its industrial production is limited due to unmature artificial diet. Insect intestinal microbiota affect host development and reproduction. The aim of this study is to understand intestinal microbiota composition of H. axyridis and screen effective probiotics on artificial diet. Considering the role of the components and composition of the diet on the structure and composition of the intestinal microbiome, four kinds of diets were set up: (1) aphid; (2) basic diet; (3) basic diet + glucose; (4) basic diet + trehalose. The gut microbiota of H. axyridis was detected after feeding on different diets.
RESULTS: Results showed that the gut microbiota between artificial diet group and aphid groups were far apart, while the basic and glucose groups were clearly clustered. Besides, the glucose group and trehalose group had one unique phylum, Cryptophyta and Candidatus Saccharibacteria, respectively. The highest abundance of Proteobacteria was found in the aphid diet. The highest abundance of Firmicutes was found in the basic diet. However, the addition of glucose or trehalose alleviated the change. In addition, the relative abundance of Enterobacter, Klebsiella, Enterobacteriaceae_unclassified, Enterobacteriales_unclassified and Serratia in the aphid group was higher than other groups. Moreover, the function of gut genes in each group also showed clear differences.
CONCLUSIONS: These results have offered a strong link between artificial diets and gut microbes, and also have provided a theoretical basis for the screening of synergistic probiotics in artificial diet.

Glyphosate is a widely-used herbicide that shows toxicity to non-target organisms. The predatory natural enemy Harmonia axyridis may ingest glyphosate present in pollen and aphid prey. The present study characterized the responses of adult H. axyridis to environmentally relevant concentrations of glyphosate (5, 10, and 20 mg/L) for one or five days. There were no obvious effects on adult H. axyridis survival rates or fecundity in response to 5 or 10 mg/L glyphosate. However, exposure to 20 mg/L glyphosate significantly reduced the survival rate and increased fecundity. Analysis of the adult H. axyridis microbiota with 16S rRNA sequencing demonstrated changes in the relative and/or total abundance of specific taxa, including Serratia, Enterobacter, Staphylococcus, and Hafnia-Obesumbacterium. These changes in symbiotic bacterial abundance may have led to changes in survival rates or fecundity of this beetle. This is the first report of herbicide-induced stimulation of fecundity in a non-target predatory natural enemy, reflecting potentially unexpected risks of glyphosate exposure in adult H. axyridis. Although glyphosate resistant crops have been widely planted, the results of this study indicate a need to strengthen glyphosate management to prevent over-use, which could cause glyphosate toxicity and threaten environmental and human health.

Evaluating the sublethal effects of insecticide is crucial for protecting and utilizing natural enemies. In this study, we determined the sublethal effects of acetamiprid and afidopyropen on Harmonia axyridis (Pallas) and explored the potential molecular mechanisms underlying these effects through transcriptomics analysis. The results showed that sublethal concentrations of acetamiprid significantly reduced the adult fecundity and longevity of F0H. axyridis and decreased the survival time and survival rate of the F1 generation. Sublethal concentrations of afidopyropen prolonged the developmental time of 4th instar larvae in the F0 generation. Additionally, acetamiprid and afidopyropen treatments significantly decreased the predation of H. axyridis. Furthermore, transcriptome sequencing analysis revealed that several P450 and UGT genes expressed differently when H. axyridis were exposed to sublethal concentrations of acetamiprid and afidopyropen, suggesting that the differential expression of detoxifying genes might be involved in the response and detoxification metabolism of acetamiprid and afidopyropen in H. axyridis. Our findings demonstrate that sublethal concentrations of acetamiprid adversely influences the development and predation of H. axyridis, while afidopyropen has limited effects on H. axyridis. These results are helpful for protecting and utilizing natural enemies and guiding the scientific use of pesticides in the field.

Maternal effects can reduce offspring susceptibility to predators by altering resource allocation to young and reproducing larger offspring. While the perception of predation risk can vary according to a prey\'s life stage, it is unclear whether maternally experienced intraguild predation (IGP) risk during different life stages influences the maternal effects of predatory insects. We investigated the influence of exposure to intraguild predators (Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae)) during the larval and/or adult stages on reproductive decisions and offspring growth in Menochilus sexmaculatus (Fabricius). Independent of the life stage, when M. sexmaculatus females experienced IGP risk, their body weight and fecundity decreased, but the proportion of trophic eggs produced increased. However, egg mass, egg clutch number, and egg clutch size were not influenced by the treatment. Next, when offspring encountered H. axyridis, mothers experiencing IGP risk during the larval and/or adult stages could increase their offspring\'s weight. Moreover, offspring in IGP environments reached a similar size as those with no-IGP environments when mothers experienced IGP risk during the larval and/or adult stages. Overall, M. sexmaculatus larval and/or adult exposure to IGP risk had no influence on egg size, but increased offspring body size when faced with H. axyridis. Additionally, mothers experiencing IGP risk during different life stages showed increased production of trophic eggs. Because IGP is frequently observed on M. sexmaculatus and favours relatively larger individuals, different stages of M. sexmaculatus express threat-sensitively to IGP risk; inducing maternal effects can be an adaptive survival strategy to defend against H. axyridis.