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Insects
Volume 16
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Insects, Volume 16, Issue 11 (November 2025) – 22 articles

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27 pages, 4104 KB  
Article
CropCLR-Wheat: A Label-Efficient Contrastive Learning Architecture for Lightweight Wheat Pest Detection
by Yan Wang, Chengze Li, Chenlu Jiang, Mingyu Liu, Shengzhe Xu, Binghua Yang and Min Dong
Insects 2025, 16(11), 1096; https://doi.org/10.3390/insects16111096 (registering DOI) - 25 Oct 2025
Abstract
To address prevalent challenges in field-based wheat pest recognition—namely, viewpoint perturbations, sample scarcity, and heterogeneous data distributions—a pest identification framework named CropCLR-Wheat is proposed, which integrates self-supervised contrastive learning with an attention-enhanced mechanism. By incorporating a viewpoint-invariant feature encoder and a diffusion-based feature [...] Read more.
To address prevalent challenges in field-based wheat pest recognition—namely, viewpoint perturbations, sample scarcity, and heterogeneous data distributions—a pest identification framework named CropCLR-Wheat is proposed, which integrates self-supervised contrastive learning with an attention-enhanced mechanism. By incorporating a viewpoint-invariant feature encoder and a diffusion-based feature filtering module, the model significantly enhances pest damage localization and feature consistency, enabling high-accuracy recognition under limited-sample conditions. In 5-shot classification tasks, CropCLR-Wheat achieves a precision of 89.4%, a recall of 87.1%, and an accuracy of 88.2%; these metrics further improve to 92.3%, 90.5%, and 91.2%, respectively, under the 10-shot setting. In the semantic segmentation of wheat pest damage regions, the model attains a mean intersection over union (mIoU) of 82.7%, with precision and recall reaching 85.2% and 82.4%, respectively, markedly outperforming advanced models such as SegFormer and Mask R-CNN. In robustness evaluation under viewpoint disturbances, a prediction consistency rate of 88.7%, a confidence variation of only 7.8%, and a prediction consistency score (PCS) of 0.914 are recorded, indicating strong stability and adaptability. Deployment results further demonstrate the framework’s practical viability: on the Jetson Nano device, an inference latency of 84 ms, a frame rate of 11.9 FPS, and an accuracy of 88.2% are achieved. These results confirm the efficiency of the proposed approach in edge computing environments. By balancing generalization performance with deployability, the proposed method provides robust support for intelligent agricultural terminal systems and holds substantial potential for wide-scale application. Full article
(This article belongs to the Special Issue Artificial Intelligence (AI) and Insect Pests Management: Securing Food Security, Human Health, and Natural Resources)
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17 pages, 3957 KB  
Article
The Oral Transfection of Spodoptera exigua (Lepidoptera: Noctuidae) Larvae via an Artificial Diet as a Strategy for Recombinant Protein Production
by María Isabel Sáez, Alba Galafat, Pablo Barranco, María Dolores Suárez, Francisco Javier Alarcón and Tomás Francisco Martínez
Insects 2025, 16(11), 1095; https://doi.org/10.3390/insects16111095 (registering DOI) - 25 Oct 2025
Abstract
Insects present extraordinary potential for obtaining recombinant proteins, both in terms of the quantity and quality of the synthesized product. This work proposes the use of artificial diets including pDNA as an oral transfection system for the Lepidoptera Spodoptera exigua. It is [...] Read more.
Insects present extraordinary potential for obtaining recombinant proteins, both in terms of the quantity and quality of the synthesized product. This work proposes the use of artificial diets including pDNA as an oral transfection system for the Lepidoptera Spodoptera exigua. It is hypothesized that oral transfection can lead to the effective expression of the reporter genes carried in plasmids. Prior to their incorporation into the artificial diet, plasmids (pCMVβ and pEGFP-N2) were protected from inactivation in the digestive tract by chitosan nanoparticulation. The survival of plasmids and their oral uptake by larvae was evaluated, as well as the persistence of pDNA in larvae throughout their ontogeny. The results confirmed that transfection occurred and that pDNA persisted during the ontogeny, even after discontinuing plasmid administration. The transcription of reporter genes was quantified by qRT-PCR, and the results indicate a dose-dependent synthesis of mRNA as the inclusion level of pDNA in diets increased. Moreover, the measurement of the biological activity of the recombinant proteins (β-galactosidase activity and green fluorescence) paralleled the results obtained for gene transcription, also dose-dependently. Therefore, effective oral transfection is feasible in S. exigua, provided that pDNA is protected against gut inactivation prior to its incorporation in artificial diets. Full article
(This article belongs to the Section Insect Molecular Biology and Genomics)
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13 pages, 2118 KB  
Article
Beyond Species Averages: Intraspecific Trait Variation Reveals Functional Convergence Under Invasion
by Zhixing Lu, Xinyu Wang, Xiang Zhang and Youqing Chen
Insects 2025, 16(11), 1094; https://doi.org/10.3390/insects16111094 (registering DOI) - 24 Oct 2025
Abstract
Biological invasions provide a unique window into community assembly. While classic theory predicts that native species must differentiate their niches to coexist with an invader, the actual outcomes under intense pressure are complex. Our study examines community reassembly under extreme pressure from the [...] Read more.
Biological invasions provide a unique window into community assembly. While classic theory predicts that native species must differentiate their niches to coexist with an invader, the actual outcomes under intense pressure are complex. Our study examines community reassembly under extreme pressure from the invasive ant Solenopsis invicta. We found that while native species do differentiate themselves from the invader, the overwhelming competition constrains this process, forcing survivors into a narrow, shared functional space. This constrained niche differentiation produces a pattern of community-level functional convergence, a process where functionally dissimilar communities become more similar under intense environmental filtering, as survivors are forced into a narrow, shared niche space. The capacity for these rapid, adaptive niche shifts is rooted in intraspecific trait variation (ITV). We also identified a dynamic feedback loop through density-dependent phenotypic plasticity in the invader. By showing how the foundational process of niche differentiation leads to a convergent outcome under extreme pressure, our work clarifies the rules of community assembly in an increasingly invaded world. Full article
(This article belongs to the Section Insect Ecology, Diversity and Conservation)
22 pages, 61965 KB  
Article
The Cercal Sensilla of the Praying Mantis Hierodula patellifera and Statilia maculata: A New Partition Based on the Cerci Ultrastructure
by Yang Wang, Xiaoqun Ding, Huan Li and Yang Liu
Insects 2025, 16(11), 1093; https://doi.org/10.3390/insects16111093 (registering DOI) - 24 Oct 2025
Abstract
Cerci function as crucial sensory organs in insects, featuring a diverse array of sensilla on their surface, analogous to those found on antennae. Using scanning electron microscopy (SEM), we characterized the ultrastructure and distribution of cercal sensilla in Hierodula patellifera (H. patellifera [...] Read more.
Cerci function as crucial sensory organs in insects, featuring a diverse array of sensilla on their surface, analogous to those found on antennae. Using scanning electron microscopy (SEM), we characterized the ultrastructure and distribution of cercal sensilla in Hierodula patellifera (H. patellifera) and Statilia maculata (S. maculata). Results show that the cerci of H. patellifera and S. maculata are highly similar, with main differences observed in the number of cercal articles and the length of cerci. The cerci of both species and sexes are composed of multiple cylindrical articles, and there is variation in the number of types of sensilla on their surface articles within sex and individuals. Females possess more cercal articles than males, and their cerci are generally longer than those of males. In both sexes of these praying mantises, four types of cercal sensilla were identified: sensilla filiformia (Sf), sensilla chaetica (Sc), sensilla campaniformia (Sca) and cuticular pore (CP), with sensilla chaetica further classified into two subtypes (ScI, ScII). Sc are widely distributed over the entire cerci, while Sf are distributed in a circular pattern on the cercal articles. While the overall distribution patterns of cercal sensilla were conserved between the sexes, significant sexual dimorphism was observed in the morphological parameters of the sensory hairs, including their quantity, length, and basal diameter. Based on distinct sensilla arrangements on the cerci, we propose a novel zoning of the cerci into four parts (I–IV), which reflects a functional gradient specialized for reproductive roles: the proximal region is enriched with robust mechanoreceptors likely involved in mating and oviposition, the central region serves as a multimodal hub for integrating courtship and mating cues, and the distal region is simplified for close-range substrate assessment. These findings highlight the adaptive evolution of cercal sensilla in relation to reproductive behaviors and provide a morphological basis for future studies on mantis phylogeny and sensory ecology. Full article
(This article belongs to the Section Insect Physiology, Reproduction and Development)
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20 pages, 2300 KB  
Article
Structural and Functional Insights into Methuselah Genes of Plutella xylostella (Lepidoptera: Plutellidae): Evolutionary Adaptations and Their Responses to Chlorantraniliprole
by Maryam Zolfaghari, Fei Yin, Samina Shabbir, Qichun Chen, Yong Xiao, Zhengke Peng, Zhen-Yu Li and Myron P. Zalucki
Insects 2025, 16(11), 1092; https://doi.org/10.3390/insects16111092 (registering DOI) - 24 Oct 2025
Abstract
G protein-coupled receptors (GPCRs) are considered the largest and most variable family of transmembrane receptors regulating physiological processes such as toxicological responses and insecticide resistance development. The present study investigated the responses of Methuselah (Mth), belonging to GPCR family B in the [...] Read more.
G protein-coupled receptors (GPCRs) are considered the largest and most variable family of transmembrane receptors regulating physiological processes such as toxicological responses and insecticide resistance development. The present study investigated the responses of Methuselah (Mth), belonging to GPCR family B in the Diamondback Moth (DBM), Plutella xylostella, to chlorantraniliprole (CAP). Genome-wide identification and phylogenetic analysis of Pxmth genes revealed their evolutionary relationships and functional classifications. Expression profiling demonstrated significant overexpression of Pxmth2 in the CAP-resistant strain. Additionally, the tertiary and secondary structures of Pxmth2 were characterized, providing insights into its functional role. Silencing Pxmth2 via RNA interference (RNAi) reduced resistance of DBM to CAP and suppressed downstream stress-associated genes (CYP6B6, CYP6B7, CYP6BF1), increasing susceptibility to the insecticide. The function of Pxmth2 was further explored using a transgenic line of Drosophila melanogaster engineered to overexpress the gene; flies overexpressing Pxmth2 exhibited a significantly increased resistance to CAP compared to controls. These findings indicate that Pxmth2 contributes to CAP resistance in DBM and highlights potential molecular targets for improving pest management strategies. Full article
(This article belongs to the Section Insect Molecular Biology and Genomics)
15 pages, 1040 KB  
Article
Distinct Modulation of Feeding Behavior in the Whitefly Vector Bemisia tabaci MED by ToCV Single-Infection Versus Synergistic Co-Infection with TYLCV
by Tianbo Ding, Hong Huang, Xiaobei Liu, Min Zhang, Jianmei Yu, Guoxu Xia and Dong Chu
Insects 2025, 16(11), 1091; https://doi.org/10.3390/insects16111091 (registering DOI) - 24 Oct 2025
Abstract
Plant viruses can significantly influence the behavior and performance of their insect vectors, with profound implications for viral epidemiology. However, studies on the effects of co-infection with multiple plant viruses on vector feeding behavior remain scarce, despite its frequent occurrence in nature and [...] Read more.
Plant viruses can significantly influence the behavior and performance of their insect vectors, with profound implications for viral epidemiology. However, studies on the effects of co-infection with multiple plant viruses on vector feeding behavior remain scarce, despite its frequent occurrence in nature and potential for altered transmission outcomes. Bemisia tabaci MED, a key vector insect, is closely linked to the rapid spread of tomato chlorosis virus (ToCV) and tomato yellow leaf curl virus (TYLCV) in China. In this study, the electrical penetration graph (EPG) technique was employed to investigate and compare the indirect (via infected plants) and direct (via viruliferous insects) effects of ToCV alone and ToCV and TYLCV co-infection on the feeding behaviors of B. tabaci MED. The results revealed that whiteflies on ToCV-infected or ToCV&TYLCV co-infected plants exhibited significantly longer non-probing durations compared to those on un-infected plants. The intracellular puncture activity of whiteflies was markedly reduced on virus-infected plants, and ToCV infection particularly shortened the duration of phloem sap ingestion. Moreover, viruliferous whiteflies (carrying ToCV or both viruses) spent less time in the intercellular pathway phase. Specifically, ToCV-viruliferous whiteflies had a shorter first-probe duration than non-viruliferous ones. The time from the first probe to the first E phase was also shorter in viruliferous whiteflies, especially in those carrying both ToCV and TYLCV. Furthermore, a significant difference was observed in the total duration of phloem sap ingestion between ToCV-viruliferous and ToCV&TYLCV-viruliferous whiteflies. These findings indicate that both ToCV infection and ToCV&TYLCV co-infection can modulate whitefly feeding behaviors through indirect and direct manners, with co-infection eliciting unique behavioral changes. These insights are valuable for elucidating the negative impact of ToCV-infected and ToCV&TYLCV co-infected tomato plants on whitefly performance, and for uncovering the mechanisms underlying the epidemics of these viruses. Full article
(This article belongs to the Special Issue Insect Transmission of Plant Viruses)
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19 pages, 3718 KB  
Article
Evaluation of the Insecticidal Potential of Lysinibacillus fusiformis Against Drosophila suzukii Larvae
by Maristella Mastore, Elisa Broggio, Davide Banfi, Ricardo A. R. Machado, Aashaq Hussain Bhat, Sadreddine Kallel, Marcella Reguzzoni, Silvia Quadroni and Maurizio F. Brivio
Insects 2025, 16(11), 1090; https://doi.org/10.3390/insects16111090 (registering DOI) - 24 Oct 2025
Abstract
The increase in the world population and consequent rise in food demand have led to the extensive use of chemical pesticides, causing environmental and health concerns. In response, biological control methods, particularly those involving microbial agents, have emerged as sustainable alternatives within integrated [...] Read more.
The increase in the world population and consequent rise in food demand have led to the extensive use of chemical pesticides, causing environmental and health concerns. In response, biological control methods, particularly those involving microbial agents, have emerged as sustainable alternatives within integrated pest management. This study highlights the potential of Lysinibacillus fusiformis as a biocontrol agent against the dipteran Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), a pest responsible for damaging soft-skinned fruits. Experimental treatments using vegetative cells, spores, and secondary metabolites of L. fusiformis on D. suzukii larvae demonstrated significant larvicidal effects, accompanied by observable changes in gut morphology under microscopy. Moreover, preliminary immunological assays showed the interference of this bacterium with the host immune system. All the results indicate the suitability of L. fusiformis for its possible integration into sustainable agricultural practices, although additional research is required to understand its applicability in the field. Full article
(This article belongs to the Special Issue Drosophila suzukii: Interactions of Microorganisms or Metazoans with the Immune System)
14 pages, 5708 KB  
Article
Integrative Taxonomic Analysis Doubles Number of Species in the Central Asian Butterfly Genus Lyela (Lepidoptera, Nymphalidae, Satyrinae)
by Vladimir A. Lukhtanov
Insects 2025, 16(11), 1089; https://doi.org/10.3390/insects16111089 (registering DOI) - 24 Oct 2025
Abstract
Lyela Swinhoe, 1908 is a small Central Asian butterfly genus, in which three species were previously recognized based on comparison of wing patterns. The present study, based on an extensive population sample across the entire range of Lyela and using integrative taxonomy methods, [...] Read more.
Lyela Swinhoe, 1908 is a small Central Asian butterfly genus, in which three species were previously recognized based on comparison of wing patterns. The present study, based on an extensive population sample across the entire range of Lyela and using integrative taxonomy methods, confirmed the monophyly of the genus and revealed the paraphyly of the most widespread species, Lyela myops sensu auct. The genus is shown to include six species, L. myops (Staudinger, 1881) (Kazakhstan, northern Kyrgyzstan, northwestern China, and southwestern Mongolia), L. tashkumirica Lukhtanov, 2024, stat. nov. (Fergana Valley in Kyrgyzstan), L. babatagi Tshikolovets, 1998, stat. nov. (southern Uzbekistan and eastern Turkmenistan), L. tekkensis (Staudinger, 1886), stat. nov. (southwestern Turkmenistan and northeastern Iran), L. macmahoni Swinhoe, 1908 (Pakistan and Afghanistan), and L. amirica Wyatt, 1961 (Afghanistan). Each of these species represents a monophyletic unity with respect to the COI gene and is separated from the other species by a distinct barcoding gap and structural differences in the male genitalia. Full article
(This article belongs to the Section Insect Systematics, Phylogeny and Evolution)
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12 pages, 5344 KB  
Article
Aphid Infestation and Predator Dynamics in Cultivated Ruta chalepensis: Evidence of Myzus persicae Adaptation and Natural Enemy Responses
by Eugênio E. Oliveira, Tarciza F. Nascimento, Leonardo S. Francesco, Thiago Svacina, César León, Carlos N. Gomes, Luis O. Viteri Jumbo, Maria José González Armijos, Karina S. Vilca Mallqui and Guy Smagghe
Insects 2025, 16(11), 1088; https://doi.org/10.3390/insects16111088 - 24 Oct 2025
Abstract
Ruta chalepensis (Rutaceae), commonly known as rue, is traditionally recognized for its medicinal applications and cultural associations with mysticism. In addition to its ethnobotanical relevance, R. chalepensis has been reported to possess insect-repellent and pesticidal properties. However, domestication and the adoption of monoculture [...] Read more.
Ruta chalepensis (Rutaceae), commonly known as rue, is traditionally recognized for its medicinal applications and cultural associations with mysticism. In addition to its ethnobotanical relevance, R. chalepensis has been reported to possess insect-repellent and pesticidal properties. However, domestication and the adoption of monoculture practices can compromise inherent plant defenses, potentially increasing susceptibility to herbivore pressure. In this context, we report, for the first time, the occurrence of infestations by the highly polyphagous aphid Myzus persicae (Sulzer) (Hemiptera: Aphididae) on R. chalepensis cultivated under Neotropical conditions. The infestations were documented in commercial rue fields located in the Zona da Mata region, Minas Gerais, Brazil, in two consecutive years. Plant material was taxonomically verified and deposited in the VIC Herbarium at the Universidade Federal de Viçosa. Aphid specimens, including both apterous and alate morphs, were examined under a stereomicroscope to confirm species identity. Field sampling revealed a notable interaction between M. persicae and native natural enemies, particularly predatory spiders. Aphid density per plant declined markedly from 14.1 to 6.2 individuals, coinciding with an observed increase in spider activity. These findings expand our understanding of M. persicae’s ecological plasticity and highlight the vulnerability of non-conventional crops to generalist pests. They also underscore the importance of implementing targeted pest surveillance and integrated pest management (IPM) approaches in underutilized or alternative cropping systems. Full article
(This article belongs to the Section Insect Pest and Vector Management)
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13 pages, 1648 KB  
Article
Ecological Balance in Unmanaged Beech Reserves: Scolytids or Their Natural Saproxylic Beetle Enemies?
by Václav Zumr, Oto Nakládal, Lukáš Bílek and Jiří Remeš
Insects 2025, 16(11), 1087; https://doi.org/10.3390/insects16111087 - 23 Oct 2025
Abstract
The accumulation of dead wood can serve as a potential source of insect pests, with scolytids being the most frequently discussed group. The aim of this study was to quantify the abundance and species composition of scolytids and their natural saproxylic beetle enemies [...] Read more.
The accumulation of dead wood can serve as a potential source of insect pests, with scolytids being the most frequently discussed group. The aim of this study was to quantify the abundance and species composition of scolytids and their natural saproxylic beetle enemies in a beech reserve. In addition, we compared the types of dead wood preferred by scolytids and their natural enemies. Beetles were sampled passively using window traps, which effectively capture the actual density of beetles within the stand. In total, 20,515 saproxylic beetles were collected; the analyses included 11 scolytids species with 3017 individuals and 51 species of natural enemies with 4976 individuals. The results revealed a significantly higher abundance of natural saproxylic beetle enemies, with no strong affiliation to specific types of dead wood. This may indicate a high mobility of natural enemies actively searching for food resources within the forest stands. In conclusion, beech reserves support high abundances of natural scolytid enemies that exceed the numbers of scolytids themselves, indicating effective natural control processes. Full article
(This article belongs to the Special Issue The Richness of the Forest Microcosmos)
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14 pages, 1615 KB  
Article
Artificial Diet Assay Screening of Candidate RNAi Effectors Against Myzus persicae (Hemiptera)
by Amol Bharat Ghodke, Stephen J. Fletcher, Ritesh G. Jain, Narelle Manzie, Neena Mitter and Karl E. Robinson
Insects 2025, 16(11), 1086; https://doi.org/10.3390/insects16111086 - 23 Oct 2025
Abstract
Aphids are sap-sucking pests that cause substantial damage to fruit and fibre crops through direct feeding and transmission of plant viruses. While chemical pesticides remain the primary method of control, their use raises concerns related to human health, environmental contamination, pesticide resistance, and [...] Read more.
Aphids are sap-sucking pests that cause substantial damage to fruit and fibre crops through direct feeding and transmission of plant viruses. While chemical pesticides remain the primary method of control, their use raises concerns related to human health, environmental contamination, pesticide resistance, and impacts on beneficial insects. As a sustainable alternative, spray-on double-stranded RNA (dsRNA) technology offers a promising approach to induce RNA interference (RNAi) in target pests. For RNAi to be effective against sap-sucking insects like the green peach aphid (Myzus persicae), it is essential to identify genes whose silencing disrupts vital physiological functions. In this study, artificial diet (AD)-based feeding assays were used to evaluate dsRNAs targeting eight genes involved in neural function, osmoregulation, feeding behaviour, and nucleic acid/protein metabolism. dsRNAs were administered individually, in combinations, or as a multi-target stacked construct. After 98 h of feeding, aphid mortality ranged from 14 to 72% (individual targets), 78–85% (combinations), and 54% (stacked construct). Transcript knockdown varied from 6.3% to ~54%, though a consistent correlation with mortality was not always observed. The gene targets and combinatorial dsRNA strategies identified in this study provide a foundation for developing RNAi-based crop protection technologies against M. persicae infestation. Full article
(This article belongs to the Section Insect Pest and Vector Management)
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8 pages, 372 KB  
Article
The Use of Disabled Insecticidal Proteins (DIPs) to Investigate the Interaction Between Aedes aegypti-Active Toxins from Bacillus thuringiensis
by Nelly Igwe and Neil Crickmore
Insects 2025, 16(11), 1085; https://doi.org/10.3390/insects16111085 - 23 Oct 2025
Abstract
A variety of pesticidal proteins derived from the bacterium Bacillus thuringiensis exhibit activity against the yellow fever mosquito Aedes aegypti and are used to control this insect vector. Several of these proteins, including Cry1Ca and Cry2Aa, additionally have activity against lepidopteran insects. Furthermore, [...] Read more.
A variety of pesticidal proteins derived from the bacterium Bacillus thuringiensis exhibit activity against the yellow fever mosquito Aedes aegypti and are used to control this insect vector. Several of these proteins, including Cry1Ca and Cry2Aa, additionally have activity against lepidopteran insects. Furthermore, the specificity of Cry2Aa has recently been shown to depend on domain I of the Cry protein, whereas it is generally recognized that domain II is the primary specificity-determining domain. This work has made use of disabled forms of three Cry proteins (Cry2Aa, Cry1Ca and Cry11Aa) and one naturally non-active protein (Cry2Ab) in an in vivo competition assay to investigate whether Cry2Aa and the dual-active Cry1Ca share a common receptor with the other pesticidal proteins. It was found that despite their differing specificities and potential modes of action, all of the Aedes-active proteins tested made use of a common receptor, although evidence is presented that Cry2Aa can use multiple receptors. When additional toxins (Cry41Aa, Cry1Aa, Cry1Ac) with no activity against this mosquito were tested, they too were found to share the same receptor, suggesting that Cry toxins may have evolved to utilize a common set of receptors in insects but that additional factors determine species specificity. Full article
(This article belongs to the Special Issue Recent Advances in Bacillus thuringiensis (Bt) Insecticides and Bt Crops: Challenges and Opportunities)
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18 pages, 5216 KB  
Article
Extraction Optimization, Multi-Dimensional Characterization, and Agricultural Application of Humic Acid from Protaetia brevitarsis Frass Derived from Straw Transformation
by Keming Chen, Qi Peng, Ziting Cheng, Lili Geng, Ting Geng, Jie Zhang and Changlong Shu
Insects 2025, 16(11), 1084; https://doi.org/10.3390/insects16111084 - 23 Oct 2025
Abstract
Agricultural straw, a massive lignocellulosic by-product, requires high-value utilization strategies, and larvae of Protaetia brevitarsis (a resource insect) can convert straw into two valuable products: insect protein and frass rich in humic acid (HA). In this study, we investigated the interactions among multiple [...] Read more.
Agricultural straw, a massive lignocellulosic by-product, requires high-value utilization strategies, and larvae of Protaetia brevitarsis (a resource insect) can convert straw into two valuable products: insect protein and frass rich in humic acid (HA). In this study, we investigated the interactions among multiple parameters affecting HA extraction efficiency and optimized the extraction process. The resulting extract was characterized by elemental analysis to determine nutrient elements, trace elements, and heavy metals; by 13C nuclear magnetic resonance (13C NMR) spectroscopy to identify the main categories of bioactive molecules; and by pot experiments to evaluate its effects on plant growth and quality. The optimized extraction conditions yielded extracts with a total organic carbon (TOC) concentration of 46.8 g/L, meeting the Chinese standard for water-soluble humic acid fertilizers (NY 1106-2010). Elemental analysis indicated that the extract was rich in trace elements, and heavy metal contents met the limitation requirements of toxic and harmful substances in fertilizers (GB 38400-2019). 13C NMR analysis revealed that the extract was enriched in aliphatic and methoxyl carbons, while pot experiments with cherry radish demonstrated that application of the extract at appropriate dosages significantly promoted plant growth and improved crop quality. These findings provide scientific support for circular agriculture and arable land protection, highlighting both their academic significance and broad application prospects. Full article
(This article belongs to the Section Role of Insects in Human Society)
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12 pages, 1099 KB  
Article
Biocontrol Potential of a Commercially Available Predator Rhyzobius lophanthae Blaisdell (Coleoptera: Coccinellidae) Against Diaphorina citri Kuwayama (Hemiptera: Liviidae)
by Gabriel Rodrigo Rugno and Jawwad A. Qureshi
Insects 2025, 16(11), 1083; https://doi.org/10.3390/insects16111083 - 23 Oct 2025
Abstract
Diaphorina citri Kuwayama is a key pest of citrus and insect vector of Huanglongbing (HLB), also known as citrus greening disease, causing significant losses in Florida and other regions. The naturally occurring effective ladybeetle predators and their impact on D. citri reduced from [...] Read more.
Diaphorina citri Kuwayama is a key pest of citrus and insect vector of Huanglongbing (HLB), also known as citrus greening disease, causing significant losses in Florida and other regions. The naturally occurring effective ladybeetle predators and their impact on D. citri reduced from years of insecticide use against this pest and are not available commercially. Additionally, most species are large-sized, while most eggs and neonates of D. citri are in hard-to-reach locations such as unopened leaves, which makes access difficult for them. We evaluated a commercially available small-sized predatory ladybeetle Rhyzobius lophanthae Blaisdell against D. citri immatures. A single adult consumed an average of 24.9 eggs and 8.7 first and second instar nymphs of D. citri within 24 h. Beetles exhibited Type II functional response against nymphs with an attack rate of 0.92 h−1 and a handling time of 0.08 h. Their consumption rate increased with nymphal density up to twenty per shoot. In the field test, beetles lived 10 days longer when confined with new shoots infested with D. citri immatures in a voile fabric sleeve cage in citrus trees every two days, versus seven days. In an open field release of R. lophanthae in a citrus orchard, these ladybeetles were found foraging in sentinel and neighboring trees infested with D. citri. The consumption rate of R. lophanthae on D. citri immatures and its survival in Florida orchards suggest its potential for biological control and Integrated Pest Management. Full article
(This article belongs to the Special Issue Improving IPM of Specialty Crop Pests and Global Food Security—2nd Edition)
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17 pages, 4227 KB  
Article
Optimizing Foliar Spray Intervals and Rates of Isocycloseram and Cyantraniliprole Plus Thiamethoxam Application on Hydrangea paniculata to Combat Adult Systena frontalis (F) (Coleoptera: Chrysomelidae)
by Shimat V. Joseph
Insects 2025, 16(11), 1082; https://doi.org/10.3390/insects16111082 - 23 Oct 2025
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Abstract
Systena frontalis (Fabricius) (Coleoptera: Chrysomelidae) is a challenging pest to manage in ornamental container nurseries, affecting over 50 plant species, particularly panicled hydrangea (Hydrangea paniculata Siebold). Because S. frontalis produces multiple generations and there is a risk of developing resistance to the [...] Read more.
Systena frontalis (Fabricius) (Coleoptera: Chrysomelidae) is a challenging pest to manage in ornamental container nurseries, affecting over 50 plant species, particularly panicled hydrangea (Hydrangea paniculata Siebold). Because S. frontalis produces multiple generations and there is a risk of developing resistance to the insecticides currently used, growers urgently seek new tools, especially new active ingredients with different modes of action or new insecticide products. Isocycloseram (Plinazolin® Technology) and cyantraniliprole + thiamethoxam (Mainspring® Xtra) are new potential insecticides for managing adult S. frontalis. However, the effective rates and application frequencies and intervals of isocycloseram and cyantraniliprole + thiamethoxam are not well understood. Therefore, the objectives of this study were to determine how rates and application intervals of these insecticides affect feeding damage when applied as foliar sprays. In laboratory assays, applying isocycloseram 1.67 SC at 59.1, 118.3, 177.4, 236.6, and 295.7 mL in 378.5 L of water, as well as cyantraniliprole + thiamethoxam at 70.9, 141.7, 212.6, and 303.3 g, reduced feeding damage compared to nontreated controls. In field trials conducted in 2024 and 2025, leaves treated with isocycloseram at 118.3 and 147.9 mL in 378.5 L of water showed significantly less damage than nontreated controls. Cyantraniliprole + thiamethoxam at 226.8 and 283.5 g in 378.5 L of water also significantly reduced the number of damaged leaves compared to nontreated controls. Two to three repeated applications of isocycloseram 1.67 SC and cyantraniliprole + thiamethoxam, administered at 7 d intervals, significantly reduced leaf damage compared to two applications at 14 d intervals. Full article
(This article belongs to the Special Issue The Efficacy of Insecticides and Botanicals Against Pests—2nd Edition)
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15 pages, 1606 KB  
Article
The Effect of Growth and Nutrition in Black Soldier Fly Larvae Fed by Hemp Seed Oil Mixed Diets
by Suttida Suwannayod, Phattawin Setthaya, Kwankamol Limsopatham and Napat Harnpornchai
Insects 2025, 16(11), 1081; https://doi.org/10.3390/insects16111081 - 23 Oct 2025
Viewed by 125
Abstract
The black soldier fly (BSF), Hermetia illucens, has gained increasing attention as a sustainable protein source for animal feed. This study investigated the effects of dietary supplementation with hemp seed oil (HSO) at 0.5–6% concentrations on the growth performance and nutritional composition of [...] Read more.
The black soldier fly (BSF), Hermetia illucens, has gained increasing attention as a sustainable protein source for animal feed. This study investigated the effects of dietary supplementation with hemp seed oil (HSO) at 0.5–6% concentrations on the growth performance and nutritional composition of black soldier fly larvae (BSFL). Larval development, survival rate, body weight, and adult longevity were evaluated under controlled conditions. In addition, chemical characterization of HSO was performed, and the proximate composition, mineral content, and amino acid profile of dried larvae were analyzed. The results indicated that HSO supplementation had no statistically significant effect on developmental time, survival rate, biomass accumulation, or adult lifespan compared to the control. Gas Chromatography–Mass Spectrometry (GC-MS) profiling of HSO revealed a wide range of bioactive compounds, including unsaturated fatty acids (UFAs), phytosterols, cannabinoids, and tocopherols. The BSFL showed high levels of fat and energy, with essential amino acids and minerals present in favorable concentrations for feed applications. The HSO improves the protein levels at the 0.5–1.0% concentration, with negative correlations at higher concentrations. The findings suggest that HSO can be integrated into BSFL diets without adverse effects on growth performance, while potentially enhancing the functional value of the larvae. This supports the feasibility of incorporating hemp by-products into insect-rearing systems to promote circular and value-added feed production. Full article
(This article belongs to the Section Insect Physiology, Reproduction and Development)
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16 pages, 703 KB  
Article
Toxicity of Hypaconitine from Aconitum coreanum (H. Lév.) Rapaics Against the Oriental Armyworm, Mythimna separata (Walker)
by Xiuwei Li, Jiaqi Xing, Meng Yang, Naiwei Chen and Yaping Liang
Insects 2025, 16(11), 1080; https://doi.org/10.3390/insects16111080 - 22 Oct 2025
Viewed by 237
Abstract
Mythimna separata (Walker) (Lepidoptera: Noctuidae) is a major migratory pest causing severe damage to cereal crops such as maize, wheat, and rice across Asia, and is also found in many parts of Oceania. With increasing insecticide resistance, botanical alternatives are urgently needed. This [...] Read more.
Mythimna separata (Walker) (Lepidoptera: Noctuidae) is a major migratory pest causing severe damage to cereal crops such as maize, wheat, and rice across Asia, and is also found in many parts of Oceania. With increasing insecticide resistance, botanical alternatives are urgently needed. This study evaluated the insecticidal potential of hypaconitine, a C19-diterpenoid alkaloid from Aconitum coreanum, against M. separata larvae. Hypaconitine exhibited significant stomach toxicity and strong antifeedant activity. It also caused pronounced growth inhibition, prolonged larval and pupal development, reduced pupation and adult emergence, induced morphological deformities, and significantly shortened adult longevity. Crucially, biochemical assays revealed sustained, time- and concentration-dependent upregulation of key detoxification enzymes—carboxylesterase (CarE), glutathione S-transferase (GST), and cytochrome P450 (CYP450)—over 72 h, indicating that hypaconitine imposes severe metabolic fitness costs rather than being readily detoxified. These effects collectively demonstrate that hypaconitine’s insecticidal efficacy arises not only from direct toxicity but also from exploiting the physiological trade-offs inherent in xenobiotic defense. Its multi-modal action—combining larvicidal, antifeedant, growth-regulatory, and metabolism-disrupting effects—presents a novel strategy for bioinsecticide development with a lower risk of resistance evolution. These findings highlight hypaconitine as a promising candidate for sustainable, integrated management of M. separata and other resistant lepidopteran pests. Full article
(This article belongs to the Section Insect Pest and Vector Management)
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28 pages, 2109 KB  
Article
Chemical Contaminants in Cerumen Samples from Ecuadorian Stingless Bees: Reporting Glyphosate, Aminomethylphosphonic Acid, and the Presence of Metals and Metalloids
by Joseline Sofía Ocaña-Cabrera, Jorge Ron-Román, Sarah Martin-Solano and Claude Saegerman
Insects 2025, 16(11), 1079; https://doi.org/10.3390/insects16111079 - 22 Oct 2025
Viewed by 292
Abstract
Stingless bee cerumen is a mixture of wax and plant resins. Foragers of stingless bees are exposed to various chemical contaminants during their plant visits and collection activities. These contaminants have the potential to be transferred into the nest. This study aimed to [...] Read more.
Stingless bee cerumen is a mixture of wax and plant resins. Foragers of stingless bees are exposed to various chemical contaminants during their plant visits and collection activities. These contaminants have the potential to be transferred into the nest. This study aimed to elucidate the existence of chemical contaminants in Ecuadorian cerumen. To this end, the following aims were established: (i) to determine and quantify glyphosate (GLY), aminomethylphosphonic acid (AMPA), some other pesticides, metals and metalloids in cerumen and (ii) to establish possible risks associated with the presence of these chemical contaminants to the health of stingless bees and humans. The quantification of chemical contaminants was conducted using gas chromatography (GC), liquid chromatography (LC), and ion chromatography (IC) coupled to mass spectrometry (MS). Glyphosate (0.02–0.2 mg/kg) and AMPA (0.028 mg/kg) were detected in four of the pooled samples (n = 14) from the northern and southern highland regions. Other pesticide traces were not detected in any cerumen samples. Metals (Cd, Cr, Pb, Ni, Sn) and metalloids (As, Sb, Se) were found in all samples, including highlands and the lower Amazon. The potential risks of exposure to glyphosate and AMPA for stingless bees and humans appear to be minimal (except for the specific conditions given for Tetragonisca angustula) and safe, respectively. It seems that cerumen may serve as an effective biomonitoring matrix for assessing the environmental health of stingless bee nests. Establishing guidelines and regulations for the safe use and handling of products derived from the stingless bee consumption is therefore imperative. Full article
(This article belongs to the Section Social Insects and Apiculture)
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27 pages, 7936 KB  
Article
Comprehensive Analysis of Cathepsin Genes in Hemiptera: Functional Characterization of the Venomous Cathepsin B from Sycanus bifidus
by Wenkai Liang, Sha Liu, Yuqin Wang, Chaoyan Wu, Wenxiu Wang and Jiaying Zhu
Insects 2025, 16(11), 1078; https://doi.org/10.3390/insects16111078 - 22 Oct 2025
Viewed by 90
Abstract
Cathepsins represent a crucial group of protein enzymes involved in insect metabolism. Within the Hemiptera order, comprising a diverse array of predatory, blood-feeding, and herbivorous species, the understanding of cathepsin types and their roles as venom components in predatory bugs remains limited. This [...] Read more.
Cathepsins represent a crucial group of protein enzymes involved in insect metabolism. Within the Hemiptera order, comprising a diverse array of predatory, blood-feeding, and herbivorous species, the understanding of cathepsin types and their roles as venom components in predatory bugs remains limited. This investigation systematically identified cathepsin genes present in Hemiptera genomes, highlighting a prevalence of cathepsin B and L, with cathepsin D exhibiting a higher gene count in the Heteroptera suborder. Examining the predatory assassin bug Sycanus bifidus, eight cathepsin genes were notably expressed in its venom glands, with the SbCAB2 gene from the cathepsin B subfamily demonstrating the highest expression in the posterior main gland, indicating its significance as a venom component. Subsequent expression and purification of the recombinant SbCAB2 protein revealed heightened hydrolytic activity (0.91 U/mg protein) compared to extracts from the anterior main gland, accessory gland, and gut. Functional assays demonstrated that SbCAB2, at lower doses (0.625–2.5 μg), can impede phenoloxidase activity in Tenebrio molitor pupal hemolymph, with a 2.5 μg dose inhibiting 86.5% of this activity, thereby preventing hemolymph melanization. Conversely, a higher dose of 10 μg led to effects akin to human placental cathepsin B, promoting melanization in T. molitor pupal hemolymph. These findings lay the foundation for further exploration of the adaptive evolution of cathepsin genes in Hemiptera and offer crucial insights into the functional role of venomous cathepsins in predatory bugs. Full article
(This article belongs to the Section Insect Molecular Biology and Genomics)
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13 pages, 1401 KB  
Article
Prey Preference of Neoseiulus californicus (McGregor) (Mesostigmata: Phytoseiidae) When Offered Two Strawberry Pests, Scirtothrips dorsalis Hood (Thysanoptera: Thripidae) and Tetranychus urticae Koch (Acari: Tetranychidae)
by Allan Busuulwa, Abigail Campos Gutiérrez and Sriyanka Lahiri
Insects 2025, 16(11), 1077; https://doi.org/10.3390/insects16111077 - 22 Oct 2025
Viewed by 220
Abstract
Mites and thrips often infest strawberry crops across the globe, with devastating outcomes for growers. In fact, Tetranychus urticae Koch and Scirtothrips dorsalis Hood are major pests of strawberries in Florida, USA, often infesting plants at the same time. Chemical control is ineffective [...] Read more.
Mites and thrips often infest strawberry crops across the globe, with devastating outcomes for growers. In fact, Tetranychus urticae Koch and Scirtothrips dorsalis Hood are major pests of strawberries in Florida, USA, often infesting plants at the same time. Chemical control is ineffective because both pests have developed resistance to many pesticides; therefore, many growers are presently utilizing predatory mites. Currently, Amblyseius swirskii Anthis-Henriot is released for S. dorsalis and Phytoseiulus persimilis Athias-Henriot for T. urticae, but this dual-release strategy is costly and can reduce effectiveness since A. swirskii can prey on P. persimilis. This study evaluated whether Neoseiulus californicus (McGregor) could provide a single practical alternative for managing both pests. We assessed its feeding preference in laboratory using no-choice and choice tests where an adult predator was offered different life stages of T. urticae and larvae of S. dorsalis. Results showed that in no-choice tests, N. californicus consumed more T. urticae eggs compared to T. urticae adults. Also, S. dorsalis larvae and T. urticae deutonymphs were consumed in similar proportions. In choice tests, feeding shifted toward mobile prey with higher consumption of S. dorsalis larvae and T. urticae deutonymphs and adults. These results demonstrate that N. californicus is a flexible predator with potential to control both pests and should be investigated further for its potential as a cost-effective tool in strawberry pest management. Full article
(This article belongs to the Special Issue Advances in the Bio-Ecology and Control of Plant-Damaging Acari)
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11 pages, 2664 KB  
Article
Effect of Solvents on the Structure of the Gut Microbiota of Honeybees (Apis mellifera)
by Kang Wang, Jinmeng Ma, Ting Ji, Haibo Zhang and Ling Yin
Insects 2025, 16(11), 1076; https://doi.org/10.3390/insects16111076 - 22 Oct 2025
Viewed by 162
Abstract
The gut microbiota of social bees is vital for host health, yet pesticide exposure can disrupt these communities. Because most active ingredients are poorly soluble, toxicological tests often use cosolvents, but their effects remain unclear. We assessed four common cosolvents (DMSO, DMF, acetone, [...] Read more.
The gut microbiota of social bees is vital for host health, yet pesticide exposure can disrupt these communities. Because most active ingredients are poorly soluble, toxicological tests often use cosolvents, but their effects remain unclear. We assessed four common cosolvents (DMSO, DMF, acetone, and Tween 80) at laboratory-relevant concentrations on honeybee survival, pollen consumption, body weight, and gut microbiota. In parallel, in vitro assays tested their impact on five dominant gut symbionts. The results showed no significant changes in survival, feeding, body weight, bacterial load, community composition, or core taxa abundance. Similarly, cosolvents did not inhibit bacterial growth in vitro. These findings demonstrate that commonly used cosolvents exert no detectable influence on honeybee physiology or gut microbiota. Although negative, this evidence is critical: it rules out cosolvents as hidden confounders, improving confidence in pesticide toxicology studies and providing essential reference data for pollinator risk assessment. Full article
(This article belongs to the Special Issue Biology and Conservation of Honey Bees)
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16 pages, 3196 KB  
Article
Transcriptional Response of ABCH Transporter Genes to Host Allelochemicals in Dendroctonus armandi and Their Functional Analysis
by Bin Liu, Jinrui Zhu and Xiaoman Ning
Insects 2025, 16(11), 1075; https://doi.org/10.3390/insects16111075 - 22 Oct 2025
Viewed by 139
Abstract
Bark beetles depend on detoxifying enzymes to counteract the defensive terpenoids produced by host trees. Insect ABC transporters play a critical role in the detoxification of insecticides and plant secondary metabolites. However, the specific functions of ABC genes in the metabolism of host [...] Read more.
Bark beetles depend on detoxifying enzymes to counteract the defensive terpenoids produced by host trees. Insect ABC transporters play a critical role in the detoxification of insecticides and plant secondary metabolites. However, the specific functions of ABC genes in the metabolism of host allelochemicals remain unclear in D. armandi. In this study, we observed that verapamil significantly enhanced the mortality of host allelochemicals in beetles, indicating that ABC transporter genes are involved in the metabolism of monoterpenes by D. armandi. We then sequenced and characterized the full-length cDNAs of three ABCH subfamily genes (DaABCH1–DaABCH3) from D. armandi. Spatiotemporal expression profiling revealed that all three genes were upregulated during developmental transitions (egg to larva and pupa to adult) and tissue-specific enrichment in detoxification-related organs (Malpighian tubules, fat body, and midgut). Additionally, DaABCH3 expression was detected in the hindgut and brain. Furthermore, DaABCH1 and DaABCH2 were significantly induced by treatment with α-pinene and limonene, whereas DaABCH3 was induced by β-pinene and limonene. Importantly, silencing DaABCH1 significantly increased mortality in adults fumigated with α-pinene and limonene. These results strongly suggest that DaABCH1 acts as a key regulator modulating D. armandi’s sensitivity to host plant allelochemicals. This finding provides a conceptual basis for developing novel control strategies against this economically significant forest pest. Full article
(This article belongs to the Special Issue Bark and Wood-Boring Insects: Past and Present Research and Essential Future Knowledge—2nd Edition)
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