Nano-enabled RNA interference (RNAi) is an emerging strategy for sustainable pest control, showing particular promise against lepidopteran insects, one of the most damaging groups to global agriculture. Conventional RNAi approaches have been hampered by instability of double-stranded RNA (dsRNA), inefficient uptake, and rapid degradation. Nanotechnology offers solutions by enhancing stability, delivery, and target specificity. Biodegradable nanocarriers—such as liposomes, chitosan-based particles, dendrimers, and polymeric systems—protect dsRNA from environmental breakdown and facilitate uptake across the gut and immune barriers of Lepidoptera. Engineered nanoparticles (NPs) enable targeted gene silencing while reducing off-target effects, improving ecological safety. The integration of nano-enabled RNAi with molecular and genomic tools supports the identification of pest-specific targets and advances environmentally compatible solutions. By lowering dependence on chemical pesticides, mitigating risks of resistance, and minimizing environmental contamination, nano-enabled RNAi aligns with green pest management principles. Despite these prospects, challenges remain, including regulatory uncertainties, production costs, ecological safety concerns, and public acceptance. Overall, nano-enabled RNAi represents a promising but still maturing approach whose success will rely on multidisciplinary progress in nanotechnology, molecular biology, and agricultural sciences.

Temperature and humidity are critical abiotic factors shaping the survival and adaptation of insect pests. However, the molecular mechanisms underlying high-temperature tolerance under contrasting humidity conditions remain poorly understood, particularly in globally invasive species such as the tomato pinworm, Tuta absoluta. Previous studies have examined individual stressors, leaving interactive thermo-hygrometric effects on gene expression and survival insufficiently resolved. Here, we assessed the contribution of cytochrome P450 genes to thermal adaptation under low- and high-humidity conditions using transcriptome profiling combined with nanocarrier-mediated RNA interference (RNAi). Third-instar larvae were exposed to high temperature under low humidity (HT-LH: 40 °C, 50% RH) or high humidity (HT-HH: 40 °C, 75% RH) for eight hours. Survival declined from 97.5% in the control to 74.16% under HT-LH and 68.33% under HT-HH conditions. Transcriptome analysis revealed extensive differential gene expression, with 464 genes upregulated and 565 downregulated in HT-LH, and 1145 upregulated and 1166 downregulated in HT-HH. Functional annotation highlighted pathways linked to metabolic regulation, proteostasis, and detoxification, including multiple cytochrome P450-associated processes. RT-qPCR confirmed the upregulation (3-5 fold) of four P450 genes (CYP6AB327, CYP6ABF1b, CYP6AE214, and CYP9A306c) under high temperature across both humidity regimes. RNAi-mediated silencing of these genes significantly reduced larval survival, demonstrating their functional role in thermal-hygrometric stress tolerance across. Cytochrome P450 genes underpin the adaptive capacity of the tomato pinworm to high-temperature stress across contrasting humidity conditions, highlighting RNAi-based disruption of P450 function as a promising avenue for sustainable pest management under climate change scenarios.

The insect gut microbiota functions as a multifunctional symbiotic system that plays a central role in host reproduction. Through the production of bioactive metabolites, gut microbes interact with host hormonal pathways, immune signaling, and molecular regulatory networks, thereby shaping reproductive physiology and fitness. This review summarizes recent advances in understanding how gut microbiota regulate insect reproduction. Accumulating evidence demonstrates that microbial metabolites contribute to nutrient metabolism and the provision of essential cofactors, modulate hormone signaling pathways involved in reproductive development, and participate in pheromone biosynthesis that affects mating behavior. These processes impact both female and male reproductive functions through coordinated interactions among metabolism, endocrine regulation, and chemical communication. In females, microbial metabolites such as short-chain fatty acids and vitamins regulate insulin/TOR and juvenile hormone signaling, promoting ovarian development, vitellogenin synthesis, and oviposition. In males, gut bacteria influence spermatogenesis, sperm motility, and pheromone production, which are critical for mating success and fertility. Overall, these findings provide a mechanistic foundation for applied strategies, including Wolbachia-based population suppression, probiotic supplementation to enhance sterile insect technique (SIT) performance, and microbial manipulation of pheromone production for pest control. In addition, dietary conditions and environmental stressors can reshape gut microbial composition and metabolic activity, leading to changes in reproductive outcomes. Furthermore, this review emphasizes the complex interactions between insect gut microbiota and reproductive physiology. Key insights include: (i) the role of microbial metabolites in regulating mating behavior, oviposition, and offspring development; (ii) the potential of microbiota-based strategies for pest control, such as Wolbachia-mediated population suppression and probiotic enhancement of SIT; and (iii) the impact of external factors, including diet and environmental conditions, on reproduction through microbiota-mediated pathways. These findings deepen our understanding of insect-microbe symbiosis and its implications for evolutionary biology and sustainable pest management. © 2026 Society of Chemical Industry.

Molting is a critical physiological process in insect development and survival, frequently targeted in both pest control strategies and insect–antagonist interactions. Parasitic wasps, in particular, manipulate host development through the injection of parasitic factors rich in Bracovirus components—agents with potential applications in sustainable pest control. Here, we investigated the interaction between the parasitoid wasp Microplitis prodeniae and its host, the fall armyworm Spodoptera frugiperda, focusing on a Bracovirus-derived parasitic factor (MpBV). Parasitism by M. prodeniae inhibited larval development, disrupted molting, and significantly reduced titers of 20-hydroxyecdysone (20E). Similar effects were observed upon MpBV injection, which led to increased mortality, decreased pupation, and suppressed 20E levels. Transcriptomic analyses revealed that 130 cuticle protein genes were downregulated in both parasitized and MpBV-injected larvae. RT-qPCR confirmed reduced expression of 20E-regulated genes at 6 and 12 h post-injection, consistent with impaired molting. Among the MpBV components, we identified an ankyrin-repeat protein (MpBVankyrin), which was detected in the hemolymph of injected larvae. Recombinant MpBVankyrin suppressed both larval development and 20E production. Yeast two-hybrid assays followed by validation demonstrated that MpBVankyrin interacts with several host proteins (SfBrC2, SfLCP22, SfRasRab, SfLCYBc1, SfCYP450V2, SfCcO, SfAnk13C, and SfGAnk), all of which were downregulated after MpBV or MpBVankyrin injection. Moreover, RNAi silencing of SfBrC2 and SfLCP22 significantly delayed larval growth. Together, these findings demonstrate that M. prodeniae disrupts host molting via MpBVankyrin, highlighting promising molecular targets for the development of molting-based pest control strategies.

Rice is a key food crop worldwide, but its yield and quality are severely constrained by insect pests. As environmental and regulatory restrictions on chemical pesticides grow, developing insect-resistant rice varieties has become a sustainable way to protect food security. This review covers recent progress in functional genomics and molecular marker mapping related to insect resistance in rice. We highlight the identification, cloning, and functional analysis of resistance genes targeting major pests, including the brown planthopper, rice gall midge, white-backed planthopper, small brown planthopper, and rice leaf roller. Several important resistance genes (such as Bph14, Bph3, and Bph29) have been cloned, and their roles in rice immunity have been clarified—covering insect feeding signal recognition, activation of salicylic acid and jasmonic acid pathways, and regulation of MAPK cascades, calcium signaling, and reactive oxygen species production. We also discuss how molecular marker-assisted selection, gene pyramiding, and transgenic techniques are used in modern rice breeding. Finally, we address future challenges and opportunities, stressing the importance of utilizing wild rice germplasm, understanding insect effector–plant immune interactions, and applying molecular design breeding to create long-lasting insect-resistant rice varieties that can withstand changing pest pressures and climate conditions.

Stomata play important roles in plant responses to phytopathogens, but less is known about their roles in plant-herbivore interactions. Herbivore damage commonly closes stomata but also elicits the release of herbivory-induced plant volatiles, important mediators of plant-herbivore interactions. Here, we examined the responses in tomato plants attacked by the invasive tomato leafminer (Phthorimaea absoluta), the larvae of which performed better on infested plants, while ovipositing adults were repelled by larvae-infested plants, which were distinct from those induced by two other tomato pests, Bemisia tabaci and Spodoptera exigua. Behavioral assays confirmed that increased releases of volatile benzaldehyde mediated P. absoluta's adult avoidance response, and large-scale glasshouse trials revealed that the induced release of benzaldehyde was sufficient to trigger long-distance dispersal of adults. The benzaldehyde release occurred through herbivory-induced stomatal openings, which were activated by the transcriptional regulation of IQ-Motif Containing Protein 1 (IQM1). P. absoluta population growth and outbreaks were amplified in glasshouse populations of wild-type plants compared to IQM1-mutated plants. P. absoluta's ability to elicit stomatal opening, as some pathogens do, may have facilitated its rapid range expansion as an invasive pest. Our study illustrates a new pattern in stomata-herbivore interactions, adding another layer of complexity to stomata-mediated plant-herbivore interactions.

Background: The diamide insecticide chlorantraniliprole is widely used in rice fields to control lepidopteran pests and is less toxic to non-target natural enemies. The egg parasitoid Trichogramma japonicum (Ashmead) is a key natural enemy deployed against these pests. However, sole reliance on T. japonicum is often insufficient for effective control and necessitates Integrated Pest Management tactics. Methods: Our study focused on the lethal and sublethal effects of chlorantraniliprole on T. japonicum, assessing its impact on growth, reproduction, oxidative stress, and physiological responses across two consecutive generations (G1 and G5). The T. japonicum adults were exposed to sublethal (LC5 = 0.00001 mg a.i. L-1, LC30 = 0.023 mg a.i. L-1) and lethal (LC50 = 0.972 mg a.i. L-1) concentrations of chlorantraniliprole over five generations (G1 to G5). Results: Exposure of T. japonicum adults to sublethal concentrations of chlorantraniliprole resulted in a marked enhancement in oviposition duration, longevity, and fecundity in the G5 generation relative to the G1 generation. The population parameters also exhibited significant enhancement at both concentrations of chlorantraniliprole (LC5 and LC30) in G5 compared with the G1 generation. Antioxidant activity (CAT and GPX enzyme activity) and the nutritional profile in T. japonicum depict a significantly higher response to the LC30 of chlorantraniliprole in both generations. Sublethal concentrations exhibited less MDA content in T. japonicum adults compared with the median lethal concentration (LC50). Conclusions: These findings support the hypothesis of hormesis. Therefore, Chlorantraniliprole-induced hormesis may favor parasitoid fitness in a consecutive generations, thereby potentially improving biological control. Also, sublethal concentrations of chlorantraniliprole could be leveraged to optimize T. japonicum mass rearing and augmentative release within IPM programs.

The continuous escalation of pest resistance leads to the ineffectiveness of most conventional pesticides, which has posed a serious threat to global food security and public health. Metabolic resistance regulated by core detoxification enzymes and penetration resistance mediated by thickened body wall/intestine are the key drivers for resistance evolution. Most existing publications focus on the functional analysis of individual target, which makes it difficult to achieve broad-spectrum resistance management. To this context, this review systematically outlines four key transcription factor-mediated metabolic signaling pathways, and analyzes the mechanism of penetration resistance mediated by body wall/intestinal thickening, aiming to identify effective RNA interference (RNAi) targets for resistance management. Subsequently, this review proposes the design/construction strategy of nano-enabled co-delivery platforms, and elaborates on their synergistic mechanisms compassing stability, foliar adhesion and plant uptake, etc. Finally, this review summarizes the application cases of nano-enabled co-delivery platforms in pest resistance management, and outlines the prospects of this technology, including multi-target coordinated interference, field adaptability improvement, etc. Overall, this work provides abundant synergistic RNAi targets for broad-spectrum resistance management, which is particularly important for design/development of multicomponent RNA nano-pesticides toward global resistant pests.

The emergence of multidrug-resistant (MDR) Pseudomonas aeruginosa in healthcare settings poses a critical global health challenge, intensified by limited therapeutic options and the environmental spread of pathogens through hospital waste. This study addresses the urgent need for alternative strategies by investigating the phylogenetic profile and antimicrobial resistance of P. aeruginosa isolated from hospital dry waste in Saidu Sharif, Pakistan, while evaluating the antibacterial and antioxidant potential of nine ethnomedicinal herbs. The isolate, identified via 16S rRNA sequencing, exhibited 99% homology to the MDR strain P. aeruginosa 610D6 reported from China, underscoring potential transboundary resistance dissemination. Antibiotic susceptibility testing revealed resistance to 14 of 21 tested agents, with retained efficacy only for imipenem, cefoperazone/sulbactam (25-mm inhibition zones), and fosfomycin (24 mm). Ethanol extracts of Punica granatum, Myrtus communis, and Olea ferruginea demonstrated remarkable antipseudomonad activity (26-mm, 26-mm, and 25-mm zone of inhibitions, respectively) and potent antioxidant properties (97.57%, 95.55%, and 96.85% DPPH scavenging). Notably, solvent polarity significantly influenced extraction yields and bioactivity, with ethanol. This work highlights the therapeutic promise of plant-derived compounds against MDR pathogens while encouraging for One Health approaches to mitigate antibiotic resistance.

Thermo-hygrometric adaptation plays a critical role in the survival of Tuta absoluta, a major pest that affects tomato crops. While general stress adaptation mechanisms are understood, the roles of key genes in thermo-hygrometric adaptation remain unclear. This study aimed to investigate the functional roles of two stress-responsive genes, HSP83 and CP7, in T. absoluta larvae under thermo-hygrometric stress. Third instar larvae were exposed to high temperature with low humidity (HT-LH: 38 degrees C, 50% RH) and high temperature with high humidity (HT-HH: 38 degrees C, 75% RH) compared to the control. Transcriptome analysis revealed differential gene expression, with 882 upregulated and 868 downregulated genes under HT-LH, and 1158 upregulated and 1069 downregulated genes under HT-HH. Gene Ontology and KEGG pathway analyses highlighted enrichment in protein processing, metabolic pathways, and cuticular structure. RT-qPCR validation confirmed significant upregulation of HSP83 (3.16-4.63-fold) and CP7 (3.55-5.85-fold) under both stress conditions. Nanocarriermediated RNAi significantly reduced the expression of HSP83 and CP7 by 0.55- and 0.53-fold, respectively. After HT-LH stress, expression levels slightly recovered but remained downregulated, and survival rates decreased to 38.3% and 31.6%, respectively. Similarly, in the HT-HH cohort, RNAi treatment led to a reduction in HSP83 and CP7 expression by 0.52- and 0.48-fold, with modest recovery post-stress. However, survival rates significantly dropped to 32.5% and 28.3%, respectively. The results suggest that CP7 and HSP83 are key players in the stress response of T. absoluta under thermo-hygrometric conditions, highlighting them as potential targets for RNAi-based pest management strategies under climate change scenarios.

Insecticides continue to play a crucial role in protecting global agriculture by preventing significant crop losses. Yet, their persistent, intensive, and often unregulated use has led to the rapid development of resistance in many pest species. The fall armyworm (Spodoptera frugiperda), a highly damaging and highly adaptable polyphagous pest, now poses one of the biggest threats to global food security. Its rapid spread and strong capacity for evolution have increased reliance on chemical control, which has contributed to rising resistance levels, environmental concerns, and economic costs. This review summarizes current knowledge on resistance mechanisms in S. frugiperda, including target-site mutations and metabolic detoxification, and offers an updated overview of global resistance patterns. In addition to chemical methods, we explore integrated and sustainable management strategies, including advanced biotechnologies such as RNA interference (RNAi), CRISPR/Cas9 gene editing, nanotechnology-based delivery systems, host plant resistance, biological control and cultural practices. Principles of Insecticide Resistance Management (IRM), such as rotating insecticides with different modes of action and systematically monitoring resistance, are discussed alongside emerging issues, including potential resistance to viral biocontrol agents. By emphasizing innovative technologies and integrated strategies, this review offers new insights into reducing the development of resistance and promoting long-term, environmentally friendly pest management.

Objective: Renal scarring is a serious long-term complication of febrile urinary tract infection (UTI) in children and may predispose affected individuals to hypertension, proteinuria, and chronic kidney disease. Early identification of children at high risk is essential for timely intervention and prevention of permanent renal damage. This study aimed to identify clinical, laboratory, microbiological, and radiological predictors of renal scarring following the first episode of febrile UTI in children. Study Design and Duration of Study: A prospective cohort study. Lady Reading Hospital from June 2023 to May 2024. Method: A total of 172 children aged 2 months to 12 years with a first documented febrile UTI completed six months of follow-up. Demographic, clinical, laboratory, microbiological, and imaging findings were recorded. Renal scarring was assessed using follow-up dimercaptosuccinic acid (DMSA) scintigraphy performed six months after the acute infection. Univariate and multivariable logistic regression analyses were performed to identify independent predictors of renal scarring. Results: Renal scarring developed in 39 children (22.7%). Children with renal scarring had significantly longer fever duration before antibiotic initiation, higher C-reactive protein levels, more frequent abnormal renal ultrasonography, acute DMSA abnormalities, and high-grade vesicoureteral reflux (all p<0.05). Multivariable analysis demonstrated that delayed antibiotic therapy beyond 48 hours (adjusted odds ratio [aOR] 3.41, 95% CI: 1.52–7.67), elevated C-reactive protein (aOR 1.18 per 10 mg/L increase, 95% CI: 1.06–1.31), abnormal renal ultrasonography (aOR 2.96, 95% CI: 1.24–7.05), acute DMSA abnormalities (aOR 5.82, 95% CI: 2.11–16.08), and high-grade vesicoureteral reflux (aOR 6.47, 95% CI: 2.18–19.21) were independent predictors of renal scarring. Conclusion: Permanent renal scarring occurred in nearly one-quarter of children after their first febrile UTI. Early diagnosis, prompt antimicrobial treatment, and risk-based imaging may facilitate early identification of high-risk children and reduce long-term renal morbidity.

Bactrocera dorsalis (Hendel, 1912), a major invasive pest, survives under extreme climates through molecular and tissue-specific cold stress adaptations. In this study, we investigated the tissue-specific impacts of cold stress on the survival and molecular response of B. dorsalis. Results showed that cold stress had a significant effect on survival rates. The Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that signaling and metabolic pathways were activated by cold stress in the head and fat body during a transcriptome analysis. Under cold stress, 184 and 365 genes were differentially expressed in the head and fat body, respectively. RNA interference (RNAi)-mediated knockdown of transposon Ty3-I Gag-Pol polyprotein (Ty3-I) and Ty3-G Gag-Pol polyprotein (Ty3-G) in the head and fat body, significantly reduced the larval survival. Relative expression analysis revealed that expression of the Ty3-I and Ty3-G Gag-Pol polyprotein was greatly reduced in the head of cold treated larvae relative to controls (dsGFP) and that the expression level of Ty3-I Gag-Pol polyprotein in the fat body was not significantly reduced by cold stress. These results highlight the tissue-specific response of Ty3-I and Ty3-G Gag-Pol polyproteins in mediating cold stress responses and aid in understanding their importance in survival and stress adaptation. Additionally, the identification of important stress-responsive genes provides a foundation for the development of RNAi-based strategies for pest control using the targeted disruption of stress adaptation gene pathways for more effective control of B. dorsalis populations.

Pesticides remain the primary method for pest management in agriculture, however, their overuse has led to resistance and raised significant concerns regarding ecological impacts and non-target organisms safety. These challenges highlight the need for sustainable, alternative pest control strategies. The sterile insect technique (SIT) has emerged as a key component of area-wide integrated pest management (AW-IPM) due to its environmental safety, species specificity, and minimal risks to humans and livestock. SIT programs typically use ionizing radiation to induce sterility in target insect populations. This review explores the scientific basis of radiation-induced sterility, focusing on its effects on germ cells, genetic material, and metabolic processes. We also examine the factors that influence the quality of sterile males-such as rearing conditions, irradiation parameters, transportation, and release strategies-all of which are critical for successful field applications of SIT. Additionally, we review recent advances in radiation-based SIT and its use in sustainable agricultural pest control and vector management. Optimizing key aspects of SIT, including rearing, irradiation, transportation, and release, as well as integrating SIT with other pest management strategies, can enhance its effectiveness. This review provides practical insights for improving SIT and outlines its potential role in the future of sustainable pest control.

Climate change increased the prevalence of heat waves, impacting insect metabolism, reproduction, development, and survival. The present study aimed to investigate the effect of heat and cold stress on the life cycle of Bactrocera dorsalis, and to identify the potential genes under thermal stress tolerance via transcriptome analysis and RNAi. Results suggest a prolonged pupal developmental duration under cold treatment (CT) and heat treatment (HT) compared to the control respectively. The mean longevity of adult was maximum in both HT and CT groups than control, but the maximum longevity was observed in CT group. The female fecundity of B. dorsalis was significantly higher in the CT compared with that of the HT and control (843.63, 753.27, and 669.27 eggs). After stress treatments, there was an increase in intrinsic and finite rates of increase in demographic parameters. After the knockdown of heat sock protein 68 (Hsp68) and insulin like-receptor (InR) using RNA interference, different body parts exhibited a divergent pattern of decreased expression levels of both genes. The expression of Hsp68 in all body parts decreased significantly after knockdown in both control and CT condition. But the lowest expression was observed in the gut (0.7-fold) as compared to dsGFP (3.35-fold) and other body parts. The InR expression was significantly reduced in all body parts at HT, while the lowest expression was observed in the head (0.9-fold). The survival rate was significantly lower under thermal stress and control conditions after gene knockdown. Taken together, our findings point towards tissue-specific and temperature-dependent effects of Hsp68 and InR expression on survival rates in B. dorsalis, implying the importance of stress and metabolic genes in designing RNAi-based pest control strategies.