Insecticide Efficacy of Green Synthesis Silver Nanoparticles on Diaphorina citri Kuwayama (Hemiptera: Liviidae).

Diaphorina citri Kuwayama (Hemiptera: Liviidae) is a vector of Liberibacter asiaticus Jagoueix et al. and Liberibacter americanus Teixeira et al., causal agents of the critical yellow dragon disease or Huanglongbing (HLB), which affects citrus production worldwide. Recently, green synthetic nanoparticles have emerged as a potential alternative to control of agricultural insect pests. The insecticide effect of silver nanoparticles (AgNPs) on 2nd instar nymphs of D. citri under laboratory and greenhouse conditions was evaluated. Mortality was recorded 24, 48, and 72 h after application on D. citri nymphs under both laboratory and greenhouse conditions. The laboratory results showed that AgNPs caused 97.84 and 100% mortality at 32 and 64 ppm, respectively, 72 h after treatment. In the greenhouse, AgNPs caused 78.69 and 80.14% mortality using 64 and 128 ppm 72 h after application. This research is the first to evaluate the green synthesis AgNPs on D. citri and are a promising strategy to control the pest.

Hydroalcoholic Extracts from Pleurotus ostreatus Spent Substrate with Nematocidal Activity against Nacobbus aberrans Phytonematode and the Non-Target Species Panagrellus redivivus.

Pleurotus ostreatus, an edible mushroom widely consumed worldwide, generates a by-product known as spent mushroom substrate (SMS). This material has demonstrated biological activity against agricultural crop pathogens. In this study, we evaluated the nematocidal effectiveness of hydroalcoholic extracts (T5, T2, AT5, and AT2) derived from SMS of P. ostreatus against (J2) of the phytonematode Nacobbus aberrans and assessed their potential toxicity towards the non-target nematode Panagrellus redivivus. Among these extracts, AT5 exhibited the highest efficacy against N. aberrans and was the least toxic against P. redivivus. Liquid-liquid partitioning yielded the AQU fraction, which showed significant nematocidal activity against J2 (75.69% ± 8.99 mortality), comparable to chitosan. The GC-MS analysis revealed the presence of several compounds, including palmitic acid, linoleic acid, and 2,4-Di-tert-butylphenol. These findings are consistent with studies confirming the antagonistic effectiveness of these compounds against phytonematodes. Additionally, all extracts exhibited toxicity against P. redivivus, with T2 being the most toxic. Our findings demonstrate that while the AT5 extract displays antagonistic effectiveness against both N. aberrans and P. redivivus, it was the least toxic among the extracts tested. Thus, SMS of P. ostreatus holds potential as a source of nematocidal compounds, which could offer significant benefits for agricultural pest control.

A new trypsin inhibitor from Centrosema plumieri effective against digestive proteases from Tribolium castaneum, an eco-friendly alternative.

Tribolium castaneum, also known as the red flour beetle, is a polyphagous pest that seriously damages agricultural products, including stored and processed grains. Researchers have aimed to discover alternative pest control mechanisms that are less harmful to the ecosystem than those currently used. We conduct the purification and characterization of a protease inhibitor from C. plumieri seeds and an in vitro evaluation of its insecticidal potential against the insect pest T. castaneum. The trypsin inhibitor was isolated from C. plumieri seeds in a single-step DEAE-Sepharose column chromatography and had a molecular mass of 50 kDA. When analyzed for interaction with different proteolytic enzymes, the inhibitor exhibited specificity against trypsin and no activity against other serine proteases such as chymotrypsin and elastase-2. The isolated inhibitor was able to inhibit digestive enzymes of T. castaneum from extracts of the intestine of this insect. Therefore, we conclude that the new protease inhibitor, specific in tryptic inhibition, of protein nature from the seeds of C. plumieri was effective in inhibiting the digestive enzymes of T. castaneum and is a promising candidate in the ecological control of pests.

Influence of Alternative Prey on the Functional Response of a Predator in Two Contexts: With and without Intraguild Predation.

In biological control, joint releases of predators and parasitoids are standard. However, intraguild predation (IGP) can occur when a predator attacks a parasitoid, potentially affecting pest control dynamics. In addition to the focal prey (FP), Trialeurodes vaporariorum, the intraguild predator (IG-predator) Geocoris punctipes can consume the parasitoid Eretmocerus eremicus (IG-prey). In this IGP context with multiple prey, an alternative prey (AP), like the aphid Myzus persicae, may influence interactions. Theory predicts that, in simple interactions, a predator's functional response (FR) to the FP changes with the presence of an AP. However, whether this holds in an IGP context is unknown. In this study, we empirically tested that prediction. Our results show that without IGP, G. punctipes exhibits a generalized FR with and without AP. Nevertheless, with IGP, the predator exhibited a Type II FR at low and high AP densities, increasing pressure on the FP and potentially favoring short-term biological control strategies. However, when 25 AP were offered, the predator's response shifted, underscoring the importance of monitoring AP densities to prevent potential disruptions in FP control. In both contexts, the increase in AP produced a handling time increase and a decrease in consumption rate. These results indicate that the theoretical prediction of the effect of AP on the FR is met only under specific conditions, and the complexity of multitrophic interactions must be considered.

Predators control pests and increase yield across crop types and climates: a meta-analysis.

Pesticides have well-documented negative consequences to control crop pests, and natural predators are alternatives and can provide an ecosystem service as biological control agents. However, there remains considerable uncertainty regarding whether such biological control can be a widely applicable solution, especially given ongoing climatic variation and climate change. Here, we performed a meta-analysis focused on field studies with natural predators to explore broadly whether and how predators might control pests and in turn increase yield. We also contrasted across studies pest suppression by a single and multiple predators and how climate influence biological control. Predators reduced pest populations by 73% on average, and increased crop yield by 25% on average. Surprisingly, the impact of predators did not depend on whether there were many or a single predator species. Precipitation seasonality was a key climatic influence on biological control: as seasonality increased, the impact of predators on pest populations increased. Taken together, the positive contribution of predators in controlling pests and increasing yield, and the consistency of such responses in the face of precipitation variability, suggest that biocontrol has the potential to be an important part of pest management and increasing food supplies as the planet precipitation patterns become increasingly variable.

Drosophila suzukii in Argentina: State of the Art and Further Perspectives.

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), commonly known as spotted-wing drosophila or SWD, is an invasive, severe, and damaging pest, which is able to inflict huge economic losses on soft thin-skinned fruits worldwide. Argentina was not excluded from the rapid invasion of this new and aggressive pest. Berries and cherries are among the most economically important fruits, showing an increasing demand from both domestic and export markets, which make necessary the application of effective and early protection measures. Although SWD is currently established almost everywhere in Argentina, the scarcity of research on and rapid regulatory actions against this pest have probably contributed to its fast spread throughout the country. In view of that, the article reviews first the current threat status of SWD in Argentina, provides summarized information on crop and non-crop host fruits, seasonal variation and population dynamics, resident natural enemy assemblages, and describes control actions implemented to date. Finally, the need to focus local control actions within an integrated national SWD management program is emphasized. The development and application of complementary eco-friendly strategies, such as Sterile Insect Technique, biological control, mass trapping, and the use of innovative lactone-derived synthetic insecticides with extremely low toxicity for SWD parasitoids, in environmentally distinguishable Argentinian regions is also highlighted.

Beauveria bassiana biogenic nanoparticles for the control of Noctuidae pests.

BACKGROUND: Biogenic metallic and oxide metal nanoparticles have potential as alternatives for several current problems in agriculture, such as the control of caterpillars which cause huge losses in the production of important crops. In the present study, capped and uncapped silver, iron oxide and titanium dioxide nanoparticles were synthesized from the filtrate of Beauveria bassiana and evaluated in regard to physico-chemical characteristics, capping composition, cytotoxicity, genotoxicity and biological activity on Helicoverpa armigera and Spodoptera frugiperda caterpillars. RESULTS: A difference in the physico-chemical parameters of the capped and uncapped nanoparticles was observed, with larger aggregation and lower stability of the uncapped. In regard to the study of the capping, the presence of functional groups of biomolecules as well as the activity of B. bassiana hydrolytic enzymes were observed. Cytotoxic effects on the tested cell lines were not observed and DNA damage levels increased with more intense effects of the uncapped nanoparticles. In regard to the biological activity against Noctuidae pests, the uncapped and capped iron oxide, and uncapped titanium dioxide nanoparticles occasioned higher mortality (76%, 60% and 51%, respectively) but no differences in LC50 were recorded. Moreover, sublethal effects were reported on Helicoverpa armigera whereas Spodoptera frugiperda showed low susceptibility to the nanoparticles. CONCLUSION: The results demonstrate that biogenic metallic and oxide metal nanoparticles might show promising effects for the control of caterpillars which cause damage on important agricultural crops. Further investigations are necessary to understand the mechanisms of action and optimize the biological activity of these new nanomaterials. © 2023 Society of Chemical Industry.

The Potential of Plant-Based Biorational Products for the Drosophila suzukii Control: Current Status, Opportunities, and Limitations.

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is an invasive species that causes serious damage to soft-skinned fruits. The use of plant-based biorational insecticides (plant extracts and essential oils) to control this pest has grown extensively. We conducted a systematic review and meta-analysis to examine the current status, trends, and perspectives of these studies, with a focus on the plant families and major compounds used as insecticides to control D. suzukii. The first article in this research field was published in 2015, and there has been exponential growth in subsequent years. Thirty-six botanical families were studied in these articles, with a prevalent interest in Myrtaceae and Lamiaceae plant species. The major constituents of these plant-based biorational molecules belong to monoterpenoids, followed by monoterpenes, benzene derivatives, and others. Geranial was the most frequent major constituent of these plant-based compounds. Our analysis revealed a few crucial consequences of the bias provided by the investigations using plant-based biorational insecticides for controlling D. suzukii. Firstly, there is a major focus on the pest species, with little or no attention paid to undesired effects on non-target beneficial organisms (e.g., pollinator bees, predators; parasitoids) and non-target pests. Secondly, the poor knowledge of how these plant-based biorational insecticides act on target and non-target organisms. Finally, there is a need to assess the efficacy of these substances under field conditions. Thus, attention is needed to address these gaps so that plant-based biorational insecticides can become a viable pest management tool for controlling D. suzukii.

First record of the major termite pest species, Reticulitermes flavipes (Isoptera: Rhinotermitidae), in Argentina.

Reticulitermes flavipes is the most invasive species in its genus and is responsible for causing significant damage to human structures in areas where it has been introduced. Although it has already become established in Chile and Uruguay, it had not previously been reported in Argentina. In this study, we report the first detection of this species in Buenos Aires, the capital of Argentina. The colony was already producing alates, and species identification was confirmed through both morphology and mitochondrial gene 16S rRNA analysis. Our results, while not conclusive, suggest that this introduction was independent from the one that occurred in Chile and Uruguay, and potentially originated from the United States. The detection of R. flavipes in Argentina is significant because it highlights the potential for this species to establish itself in new regions and underscores the need for future research on and control of R. flavipes in this country.

Production of Escovopsis weberi (Ascomycota: Hypocreales) Mycelial Pellets and Their Effects on Leaf-Cutting Ant Fungal Gardens.

The maintenance of the symbiosis between leaf-cutting ants and their mutualistic fungus Leucoagaricus gongylophorus Singer (Moller) is vital for the survival of both species. The specialist fungal parasite Escovopsis weberi Muchovej & Della Lucia is a threat to this symbiosis, causing severe damage to the fungal garden. Mycelial pellets are resistant fungal structures that can be produced under laboratory conditions. These structures were studied for use in biological pest control, but the production of mycelial pellets has not previously been documented in Escovopsis. One of the aims of this study was to induce Escovopsis weberi to produce mycelial pellets and investigate the potential of these pellets for the control of leaf-cutting ants. We compared the pathogenicity of Escovopsis weberi mycelial pellets and conidia against mini-colonies of Acromyrmex subterraneus subterraneus Forel when applied in the form of baits. Worker ants were able to distinguish mycelial pellets from conidia, as baits with mycelial pellets were more attractive to workers than those with conidia, causing a greater negative impact on colony health. All types of baits containing Escovopsis weberi influenced the foraging activity but only treatments with viable fungal propagules resulted in an increase in the quantity of waste material, with a significant negative impact on the fungal garden biomass. The results provided novel information regarding Escovopsis recognition by worker ants and differences between conidia and mycelial pellet dynamics in leaf-cutting ant colonies, with new perspectives for the biological control of these important pests.

Research Trends, Biases, and Gaps in Phytochemicals as Insecticides: Literature Survey and Meta-Analysis.

A 76-year literature survey and meta-analyses were carried out to recognize the trends, biases, and knowledge gaps of studies focusing on major groups of compounds of botanical origin, or phytochemicals, as insecticides. The survey found that the main phytochemicals prospected as insecticides belong to the following major chemical groups: terpenoids, terpenes, and carbonyl, all of which were tested, mainly against beetles (Coleoptera), caterpillars (i.e., larvae of Lepidoptera), and mosquitoes and other flies (i.e., Diptera). These studies are burgeoning at an exponential rate, with an evident focus on mortality endpoint estimates, but they are also neglecting sublethal assessments. China and India in Asia, as well as Brazil in the Americas, were responsible for most studies. The majority of the papers used stored grain insects as experimental models, which limits the applicability and representativeness of the findings. As a result, the main modes of exposure tested were fumigation and contact, which leads to the prevalence of estimates of lethal concentration in these studies. Therefore, a broader range of insect species deserves testing, with suitable modes of exposure identifying and characterizing the main molecules responsible for the insecticidal activity, which is seldom performed. Attention to these needs will circumvent current biases and allow the recognition of the main patterns of association between the origin and structure of phytochemicals and their insecticidal effects.

Azadirachtin-based insecticide impairs testis morphology and spermatogenesis of the southern armyworm Spodoptera eridania (Lepidoptera: Noctuidae).

BACKGROUND: In the search for alternative tools for integrated pest management, azadirachtin, a botanical insecticide, has been used with the most promising activity against Spodoptera spp., but the mechanism of cytotoxicity on reproductive organs remains unclear. Spodoptera eridania (Stoll, 1782) is a polyphagous pest with great economic importance that has become an important target to elucidate the action of azadirachtin on the reproductive organs of insect pests, helping to understand the deleterious effects caused by its exposure. This study evaluated the effects of chronic exposure to azadirachtin on the morphology and ultrastructure of S. eridania larval testes as well as larval development. RESULTS: Azadirachtin exposure (6 or 18 mg a.i. L-1 ) caused a progressive increase in cumulative mortality and reduced gain in body mass after 5 days. Testicular structure indicated a reduction in their size with internal morphological changes such as spermatogonia, spermatogonial, spermatocytes and spermatid cysts in degeneration. The occurrence of cell death in germ and somatic cells was evidenced by the TUNEL technique. Electron microscopy revealed changes in cystic cells, such as cytoplasmic membrane rupture and cytoplasmic vacuolization. Chromatin compaction, changes in the rough endoplasmic reticulum and Golgi complex cisternae were observed in germ cells. Apoptotic bodies occurred between germ cell cysts. CONCLUSION: Azadirachtin damaged the testes of S. eridania larvae, and these changes compromised spermatogenesis and consequently the development of the reproductive potential of this specimen, making azadirachtin a promising botanical insecticide for application in integrated pest management programs. © 2022 Society of Chemical Industry.

Entomopathogenic Fungi-Mediated AgNPs: Synthesis and Insecticidal Effect against Plutella xylostella (Lepidoptera: Plutellidae).

The insect Plutella xylostella is known worldwide to cause severe damage to brassica plantations because of its resistance against several groups of chemicals and pesticides. Efforts have been conducted to overcome the barrier of P. xylostella genetic resistance. Because of their easy production and effective insecticidal activity against different insect orders, silver nanoparticles are proposed as an alternative for agricultural pest control. The use of entomopathogenic fungi for nanoparticle production may offer additional advantages since fungal biomolecules may synergistically improve the nanoparticle's effectiveness. The present study aimed to synthesize silver nanoparticles using aqueous extracts of Beauveria bassiana, Metarhizium anisopliae, and Isaria fumosorosea isolates and to evaluate their insecticidal activity against P. xylostella, as innovative nano-ecofriendly pest control. The produced silver nanoparticles were evaluated by measuring the UV-vis spectrum and the mean particle size by dynamic light scattering (DLS). I. fumosorosea aqueous extract with 3-mM silver nitrate solution showed the most promising results (86-nm mean diameter and 0.37 of polydispersity). Scanning electron microscopy showed spherical nanoparticles and Fourier-Transform Infrared Spectroscopy revealed the presence of amine and amide groups, possibly responsible for nanoparticles' reduction and stabilization. The CL50 value of 0.691 mg mL-1 was determined at 72-h for the second-instar larvae of the P. xylostella, promoting a 78% of cumulative mortality rate after the entire larval stage. From our results, the synthesis of silver nanoparticles mediated by entomopathogenic fungi was successful in obtaining an efficient product for insect pest control. The I. fumosorosea was the most suitable isolate for the synthesis of silver nanoparticles contributing to the development of a green nanoproduct and the potential control of P. xylostella.

Analysis of glutathione-S-transferases from larvae of Galleria mellonella (Lepidoptera, Pyralidae) with potential alkaloid detoxification function.

The greater wax moth, Galleria mellonella, is a global pest for beehives, doing damage in the larval stage. Although a significant number of studies have reported on larvae and adults, to date no effective pest control has been implemented. In this study, we tested larval resistance to alkaloids from Berberis microphylla, and the objective was to identify enzymes that participate in alkaloid detoxification through enzymatic assays, bioinformatics analysis and qRT-PCR. Findings suggest glutathione-S-transferases (GSTs), from an increased metabolic mechanism, are responsible for alkaloid detoxification rather than cytochrome P450 (CYP), carboxylesterases (CarE). A bioinformatics analysis from transcriptome data revealed 22 GSTs present in both G. mellonella larvae and adults. The qRT-PCR experiments corroborated the presence of the 22 GSTs in larvae, where GST8 and GST20 stood out with the highest expression after berberine treatment. Structural information around GST8 and GST20 suggests that GST8 could bind berberine stronger than GST20. These findings represent an important advance in the study of detoxification enzymes in G. mellonella, expanding the role of delta-class GSTs towards alkaloids. Likewise, GST inhibition by alkaloid analogs is proposed in the framework of integrated pest management strategies.

Insights Into Chemosensory Proteins From Non-Model Insects: Advances and Perspectives in the Context of Pest Management.

Nowadays, insect chemosensation represents a key aspect of integrated pest management in the Anthropocene epoch. Olfaction-related proteins have been the focus of studies due to their function in vital processes, such ashost finding and reproduction behavior. Hence, most research has been based on the study of model insects, namely Drosophila melanogaster, Bombyx mori or Tribolium castaneum. Over the passage of time and the advance of new molecular techniques, insects considered non-models have been studied, contributing greatly to the knowledge of insect olfactory systems and enhanced pest control methods. In this review, a reference point for non-model insects is proposed and the concept of model and non-model insects is discussed. Likewise, it summarizes and discusses the progress and contribution in the olfaction field of both model and non-model insects considered pests in agriculture.

Nanoparticles Loaded with Essential Oil from Zanthoxylum riedelianum Engl. Leaves: Characterization and Effects on Bemisia tabaci Middle-East Asia Minor 1.

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) Middle-East Asia Minor 1 is a major pest of agricultural production systems. It is controlled by synthetic insecticides. Essential oils are promising eco-friendly alternatives. This study developed and characterized nanoparticles loaded with essential oils of Zanthoxylum riedelianum Engl. (Rutaceae) leaves and evaluated their potential for B. tabaci management. The essential oil exhibited an average yield of 0.02% (w w-1) and showed as major components γ-elemene (24.81%), phytol (18.16%), bicyclogermacrene (16.18%), cis-nerolidol (8.26%), and D-germacrene (6.52%). Characterization of the nanoparticles showed a pH between 4.5 and 6.7, a zeta potential of approximately - 25 mV, particle-size distribution ranging from 450 to 550 nm, and encapsulation efficiency close to 98%. The nanoencapsulation was an efficient process that provided photostability against photodegradation. Bioassays with crude and nanoencapsulated essential oils significantly reduced the number of nymphs and eggs of B. tabaci, with the best results observed at concentrations of 5 and 2% (v v-1). Our results demonstrated that essential oils from Z. riedelianum can be nanoformulated resulting in a stable product while maintaining their biological activity against B. tabaci Middle-East Asia Minor 1.

Overview of paratransgenesis as a strategy to control pathogen transmission by insect vectors.

This article presents an overview of paratransgenesis as a strategy to control pathogen transmission by insect vectors. It first briefly summarises some of the disease-causing pathogens vectored by insects and emphasises the need for innovative control methods to counter the threat of resistance by both the vector insect to pesticides and the pathogens to therapeutic drugs. Subsequently, the state of art of paratransgenesis is described, which is a particularly ingenious method currently under development in many important vector insects that could provide an additional powerful tool for use in integrated pest control programmes. The requirements and recent advances of the paratransgenesis technique are detailed and an overview is given of the microorganisms selected for genetic modification, the effector molecules to be expressed and the environmental spread of the transgenic bacteria into wild insect populations. The results of experimental models of paratransgenesis developed with triatomines, mosquitoes, sandflies and tsetse flies are analysed. Finally, the regulatory and safety rules to be satisfied for the successful environmental release of the genetically engineered organisms produced in paratransgenesis are considered.

RNAi in Piezodorus guildinii (Hemiptera: Pentatomidae): Transcriptome Assembly for the Development of Pest Control Strategies.

Red-banded stink bug Piezodorus guildinii (P. guildinii) has been described as the most damaging stink bug regarding soybean crops, leading to seed injury, low germination percentages, and foliar retention, at low population densities. In recent years, RNA interference (RNAi), a conserved eukaryote silencing mechanism has been explored to develop species-selective pesticides. In this work, we evaluated RNAi in P. guildinii to develop new pest-control strategies. For this, we assembled and annotated a P. guildinii transcriptome from a pool of all developmental stages. Analysis of this transcriptome led to the identification of 56 genes related to the silencing process encompassing siRNA, miRNA, and piRNA pathways. To evaluate the functionality of RNAi machinery, P. guildinii adults were injected with 28 ng/mg of body weight of double stranded RNA (dsRNA) targeting vATPase A. A mortality of 35 and 51.6% was observed after 7 and 14 days, respectively, and a downregulation of vATPase A gene of 84% 72 h post-injection. In addition, Dicer-2 and Argonaute-2 genes, core RNAi proteins, were upregulated 1.8-fold 48 h after injection. These findings showed for the first time that RNAi is functional in P. guildinii and the silencing of essential genes has a significant effect in adult viability. Taken together, the work reported here shows that RNAi could be an interesting approach for the development of red-banded stink bug control strategies.

Chemical control of Acromyrmex lundi (Hymenoptera: Formicidae) in anthropic ecosystems.

BACKGROUND: Leaf-cutting ants are pests in agriculture, forests and pasture and, usually, controlled with toxic baits. The management and control of leaf-cutting ants is complex as a consequence of its sociability and behavior. We tested three toxic baits with abamectin, fipronil and sulfluramid to control colonies of Acromyrmex lundi Mayr, 1865 (Hymenoptera: Formicidae), in two seasons: 'Spring 2020' and 'Summer 2020/21', in Argentina. RESULTS: The percentage of effective bait carried in the spring was similar between treatments with 80%, 80% and 90% for those with abamectin, fipronil and sulfluramid. The ant flow of A. lundi decreased after toxic bait applications compared to the control through time in all treatments. This activity was higher in the spring than in the summer. The baits reduced the foraging activity of A. lundi from 5 to 96 days after application (DAA). Abamectin and fipronil baits controlled 50% and 67% of the nests in the spring, and 20% and 43% at 96 DAA in the summer. Sulfluramid was the best treatment, killing 90% of A. lundi nests. CONCLUSION: The baits decrease the foraging activity of A. lundi until 96 DAA, with total or partial death of its colonies varying between treatments. The ant symptoms with the sulfluramid bait appeared faster than with fipronil and may be a result of the low level of carry-over or differences in the chemical quality of the active ingredients. © 2022 Society of Chemical Industry.

Inhibition constant and stability of tripeptide inhibitors of gut trypsin-like enzyme of the soybean pest Anticarsia gemmatalis.

Insects overcome the action of natural protease inhibitors (PIs) due to evolutionary adaptations through endogenous proteolysis and reprogramming proteases. Insect adaptations complicate the formulation of IP-based crop protection products. However, small peptides designed based on the active site of enzymes have shown promising results that could change this scenario. GORE1 and GORE2 are designed tripeptides that reduce the survival of Anticarsia gemmatalis when ingested orally. In this article, the stability and ability of the peptides to bind trypsin-like enzymes of A. gemmatalis were evaluated by molecular dynamics (MD) simulations. The ability of the peptides to inhibit trypsin-like enzymes in vivo was compared with the SKTI protein by feeding A. gemmatalis larvae at different concentrations, followed by an inhibition persistence assay. During the MD simulation of enzyme-ligand complexes, both peptides showed a small variation of root-mean-square deviation and root-mean-square fluctuation, suggesting that these molecules reach equilibrium when forming a complex with the trypsin-like enzyme. Furthermore, both peptides form hydrogen bonds with substrate recognition sites of A. gemmatalis trypsin-like enzyme, with GORE2 having more interactions than GORE1. Larvae of A. gemmatalis exposed to the peptides and SKTI showed a similar reduction in proteolytic activity, but the persistence of inhibition of trypsin-like enzyme was longer in peptide-fed insects. Despite their size, the peptides exhibit important active and substrate binding site interactions, stability during complex formation, and steadiness effects in vivo. The results provide fundamental information for the development of mimetic molecules and help in decision-making for the selection of delivery methods for larger-scale experiments regarding similar molecules.