IntroductionLarval diet plays a crucial role in mosquito development and fitness by influencing both larval and adult traits. During the aquatic stage, mosquitoes acquire essential nutrients required for successful metamorphosis and adult emergence. Interactions between larval nutrition and bacterial endosymbionts such as Wolbachia further influence mosquito development, with important implications for field-based vector control strategies.MethodsWe evaluated the effects of four larval diets on fitness characteristics of Wolbachia-transinfected and uninfected Ae. aegypti strains. The diets tested were: LD1 (fish feed), LD2 (laboratory rodent diet), LD3 (mushroom powder), and LD4 (dog biscuit plus brewer’s yeast). We assessed wing length and body weight in Wolbachia-transinfected strains (wMel and wAlbB) and uninfected strains across two generations of Ae. aegypti reared under each larval diet regimen. Additionally, Wolbachia density was measured in the transinfected Ae. aegypti strains using real-time PCR.ResultsFemale wing length was significantly influenced by the interaction between diet, Wolbachia strain, and generation (Diet × Strain × Generation: F = 3.98, P = 0.0044), together with significant Diet × Strain (F = 3.05, P = 0.0135. In contrast, male wing length was primarily affected by diet (F = 14.11, P < 0.001) and the Diet × Generation interaction (F = 5.58, P = 0.0019). Male body weight also showed a significant three-way interaction among diet, strain, and generation (F = 8.95, P < 0.001), indicating strain- and generation-specific dietary responses. Female body weight exhibited significant Diet × Strain (F = 3.43, P = 0.0069 and Strain × Generation (F = 7.79, P = 0.0012) interactions. Overall, larval diet was the primary determinant of mosquito fitness, with its effects modified by Wolbachia infection status and generation.DiscussionThis study emphasizes that larval diet significantly influences the fitness of both Wolbachia-transinfected and uninfected Aedes aegypti strains. LD1 and LD4 showed improved wing development and fitness compared to LD2 and LD3, with sex-specific effects observed. Based on the analysis, LD1 is recommended for routine mass rearing of Wolbachia trans-infected mosquitoes. These findings highlight the need for future studies on cost-effective, locally available diet formulations to optimize mosquito fitness for vector control strategies.
IntroductionHouse dust mites (HDMs) are major indoor allergens worldwide, with Dermatophagoides farinae being one of the most prevalent species. Temperature is a key driver of mite development, but the thermal biology of HDMs remains poorly characterized. Despite the medical importance of house dust mites, comprehensive thermal performance data across ecologically relevant temperature ranges remain limited, particularly for upper thermal thresholds that will become increasingly relevant under climate warming.MethodsWe evaluated the development, survival, and fecundity of D. farinae under five constant temperatures (15, 20, 25, 30, and 35°C) at 75 ± 5% relative humidity using the age-stage, two-sex life table framework.ResultsDevelopmental duration decreased significantly from 15 to 30°C but increased at 35°C due to thermal inhibition. The shortest pre-adult development time was 19.14 days at 30°C, while the longest was 68.98 days at 15°C. Egg-to-adult survival was highest at 25°C (88.6%) and lowest at 35°C (35.7%). The net reproductive rate (R0) peaked at 25°C (33.96 offspring per female), whereas the intrinsic rate of increase (rm) was highest at 30°C (0.0598 day−1), reflecting the differential contributions of fecundity versus developmental speed to population performance. D. farinae exhibits optimal development at 25–30°C, with severe thermal stress above 35°C.DiscussionThese baseline thermal performance data provide essential parameters for population models aimed at assessing climate change impacts. Our results suggest that climate warming may initially increase HDM populations in temperate regions but could restrict their distribution in areas where thermal thresholds are exceeded. These findings provide baseline data for assessing allergen exposure risks under future climate scenarios.
RNA interference (RNAi) is an eco-friendly strategy for pest management, with double-stranded RNA (dsRNA) as the core functional component. In this study, three RNAi target genes (Ubx, wupA and Dpp) with strong lethal effects on Apolygus lucorum were screened via microinjection. The 7-day cumulative mortalities were 56.67 ± 3.33% for dsUbx, 94.44 ± 1.11% for dswupA and 92.22 ± 1.11% for dsDpp. We optimized dsRNA sequences by removing conserved sequences in non-target organisms based on homology alignment and off-target risk analysis. The optimized fragments dswupA-OTE and dsDpp-OTE still exhibited high insecticidal activity, with 7-day cumulative mortalities of 77.78 ± 2.94% and 70.00 ± 1.93%, respectively. We also evaluated the effects of dsRNA length and target sites on RNAi efficiency and screened potent short dsRNA fragments. Novel artificially recombinant dsRNAs were constructed by assembling effective short fragments from different genes, which retained strong insecticidal activity despite shorter sequence length. This study verifies the feasibility of multi-target recombinant dsRNA for pest control and provides a theoretical basis for developing multi-gene RNAi technologies against A. lucorum.
Varroa destructor is a major global pest of the European honey bee, and RNA interference (RNAi) has emerged as a promising technology for the development of effective control tools. However, the efficacy of RNAi-based approaches remains highly variable, due in part to the rapid degradation and inefficient delivery of exogenously applied double stranded RNA (dsRNA) This study investigated whether advanced dsRNA molecular designs could improve dsRNA stability, processing, and biological outcomes in the honey bee-Varroa mite pathosystem. Loop-ended dsRNA (ledRNA) constructs incorporating either G-U wobble base-pairing or asymmetric bulge modifications were designed to target two Varroa genes: the chitin-binding protein Peritrophin-A-like (Pero) and the neuropeptide crustacean hyperglycemic hormone (CHH). Stability assays, immersion and indirect feeding bioassays, and small RNA sequencing were used to evaluate dsRNA persistence, RNAi activity, and biological efficacy compared to conventional dsRNA. The advanced designs exhibited double the stability in sucrose feeding solutions and within adult bees compared with conventional dsRNA. In immersion and indirect feeding bioassays, these designs produced variable but enhanced effects on target gene silencing (~36-86% reduction) and mite mortality (~32-56% increase) when compared with conventional dsRNA. Small RNA sequencing revealed effective processing of the advanced dsRNA designs, with complete siRNA coverage of the target region, 22-24 nt peak size classes and antisense strand bias. Together, these results demonstrate that advanced dsRNA molecules have increased stability, effective processing, and strong efficacy in V. destructor under laboratory conditions, supporting further investigation of structurally optimized dsRNA designs for field hive applications.
The salivary gland (SG) is the final barrier for Plasmodium transmission to humans but remains comparatively understudied relative to the midgut microbiome. This review synthesizes current knowledge on SG microbiome acquisition routes, composition, and functional significance. Acquisition may occur via larval filter feeding, vertical (egg smearing), transstadial, or horizontal transmission during blood feeding, though their relative contributions are unknown. Compositional studies show Gram-negative genera Serratia, Elizabethkingia, Acinetobacter, Pseudomonas, and Asaia predominate; Plasmodium infection correlates with increased Serratia and decreased Elizabethkingia abundance. While immune-related genes (e.g., cecropins, defensin, GNBP, SRPN6) expressed in the SG may be modulated by resident bacteria, direct evidence of their effect on sporozoite invasion remains lacking. Gram-negative bacteria trigger Toll, Imd, and JAK-STAT pathways, but emerging evidence suggests the SG may mount a distinct, locally independent immune response compared to the systemic pathway. Paratransgenesis using Asaia shows promise, yet SG-targeted effector delivery remains untested. Ecological pressures common in West Africa, including agricultural pesticides, insecticide resistance, and larval water contamination, may influence mosquito-associated bacteria, but no studies explicitly link these to the SG microbiome. Significant knowledge gaps persist, notably the absence of field studies in high-burden regions like Nigeria and the lack of experimental manipulation to establish causality. Addressing these priorities is critical to determine whether the SG microbiome can be exploited as a transmission-blocking target.
Many phytophagous insects develop within discrete and non-renewable resources, such as fruits, making host choice a critical determinant of offspring performance. In tephritid fruit flies, host fruit quality can strongly influence development, survival, and adult reproductive traits, yet no single host may optimize all life-history parameters. Here, we evaluated the effects of six host fruit species—native and exotic—on larval development and adult performance of the South American fruit fly Anastrepha fraterculus, a highly polyphagous and economically important pest. Under controlled laboratory conditions, we quantified developmental time, pupal survival, adult size, ovary development, male sexual maturation, and starvation resistance. Host species significantly affected all measured traits, revealing pronounced trade-offs among life-history parameters. Guava, a native host, supported the fastest larval development but produced smaller adults with reduced ovary development and lower starvation resistance. Feijoa, another native host, behaved similarly to guava, although pupal mortality was higher and sexual maturation faster than in guava. Peach and plum consistently yielded larger adults with well-developed ovaries and higher overall performance, whereas loquat generally resulted in poorer outcomes across several traits. Grapefruit induced prolonged larval development but promoted early male sexual maturation. Heatmap and PCA analyses confirmed that no single host maximized performance across all traits. These results challenge the assumption that native hosts necessarily confer superior fitness and suggest a decoupling between host use in nature and host suitability under laboratory conditions. Our findings highlight the complexity of host–insect interactions in polyphagous fruit flies and suggest that different host species may be selectively advantageous depending on ecological context and demographic needs. Understanding these trade-offs is essential for predicting population dynamics and improving integrated management strategies for A. fraterculus.
IntroductionMass rearing for the Sterile Insect Technique (SIT) may induce transcriptomic changes that influence post-release performance. We compared head transcriptomes of two mass-reared Anastrepha ludens strains, Bisexual and Tap-7, with wild individuals associated with their preferred natural host, sweet orange.MethodsRNA-seq data were analyzed to assess transcriptomic differentiation, differential gene expression, Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and chemosensory gene expression profiles among the three groups.ResultsPrincipal component analysis clearly separated the mass-reared strains from wild host-associated individuals. More than 1,300 differentially expressed genes were identified per strain, with gene repression predominating and enrichment of functions related to translation, cellular energy metabolism, RNA processing, and metabolic regulation. Both mass-reared strains shared signatures of metabolic adjustment but also displayed strain-specific patterns. The Bisexual strain showed signatures associated with stress response, immunity, and mitochondrial quality control, whereas Tap-7 showed enrichment of pathways related to calcium regulation, secretion, detoxification, and folate metabolism. Both strains also exhibited reorganization of chemosensory gene expression, including conservation of receptors and co-receptors such as Ir25a, Ir68b, Ir85a, Or74a, and SNMP1, together with convergent downregulation of odorant-binding proteins, particularly Obp56a-like and members of the Obp99 clade.DiscussionThese results indicate that mass-reared A. ludens strains exhibit a distinct head transcriptomic state relative to wild host-associated flies. The shared and strain-specific expression patterns suggest that mass rearing may be associated with metabolic and sensory changes relevant to post-release performance. The identified sensory and metabolic markers provide candidates for future behavioral, electrophysiological, and physiological evaluation of fruit fly strains used in SIT programs.
IntroductionIn crop pest control, microbial bioinsecticides are considered promising for overcoming the critical limitations of traditional chemical pesticides regarding environmental and health impact. In this study, we report a strain of Pantoea agglomerans (ELIV) lethal to black bean aphid Aphis fabae: we investigated the effects of this bacterium on aphids and host plants to test its potential as a natural insecticide. Methods and ResultsWe found that the entomopathogenic properties of P. agglomerans ELIV occur only through ingestion via oral infection assays, with no pathogenicity by contact observed via spraying tests. The bacterium does not appear to circulate within the plant after watering, remaining at the root level and having only slight effects on tested plant growth parameters. Genome analysis of P. agglomerans ELIV reveals its richness in genes encoding virulence factors, particularly iron-chelating siderophores, possibly explaining its ability to rapidly kill infected insects. DiscussionOverall, our results present the strengths and limitations of the species P. agglomerans for the development of new bioinsecticides. Although the use of P. agglomerans may be of limited interest for controlling piercing-sucking insects as they feed internally, it may be relevant against chewing insect pests, e.g. lepidopteran caterpillars and beetles.
Malaria remains one of the most devastating infectious diseases globally, with Anopheles gambiae serving as the principal vector across sub-Saharan Africa. The effectiveness of traditional vector control methods is currently under threat due to the rapid rise of pesticide resistance, which calls for the discovery of new molecular targets. The GTP-dependent GTPase known as Eukaryotic Elongation Factor 2 (EF-2; encoded by AGAP009441 in An. gambiae) is essential for cellular viability because it catalyzes the translocation stage of ribosomal protein production. EF-2 was identified as an important gene in An. gambiae by machine learning-based computational screening, and RNA interference (RNAi)-mediated knockdown experimentally verified that EF-2 silencing dramatically shortens mosquito longevity (p < 0.0001). Despite the fact that EF-2 and its human homolog share about 79% of the same protein sequence, species-specific structural characteristics may provide opportunities for selective targeting. Additionally, a comparative dN/dS analysis was carried out on EF-2 across Anopheles species using the HyPhy Datamonkey platform. The dN/dS results revealed high evolutionary conservation (ω = 0.0198) and are consistent with purifying selection, but not with positive selection. The current understanding of EF-2 biology, its validation as a vector control target, suitable intervention modalities like RNA interference and small-molecule inhibition, and the major issues of selectivity, delivery, and ecological safety that need to be resolved prior to translational application are all summarized in this review. EF-2 is a promising but understudied contender whose complete potential in integrated malaria vector control necessitates immediate and ongoing research.
Spotted lanternfly, Lycorma delicatula (White) is a planthopper (Hemiptera: Fulgoridae) that is native to China and invasive elsewhere in Asia and North America. The insect feeds on several plants and is an emerging pest of apples, grapes, walnuts, and other hardwoods. Forecasts of the ultimate geographic distribution for L. delicatula in Asia and North America, particularly its northern latitudinal limits, are uncertain because of limited information about the cold tolerance of this insect. Cooling rate can affect supercooling point, the temperature at which bodily fluids begin to freeze and a common reference point for insect cold tolerance studies. This study measures changes in supercooling point of overwintering L. delicatula eggs in response to different cooling rates. Overwintering egg masses were collected from two field locations in Virginia, USA in November and December 2024. Supercooling points of individual eggs were measured with contact thermocouple thermometry at cooling rates of 0.033, 0.5, and 1 °C/min. The distribution of supercooling points changed with different cooling rates and were negatively related to the cooling rate. At a cooling rate of 0.033 °C/min, SCPs ranged from -28.4 to -22.9 °C and, at 1 °C/min, from -29.2 to -24.7 °C. If supercooling points reflect lethal temperatures for L. delicatula, relatively rapid cooling may slightly overestimate the cold tolerance of the insect and inflate the estimated geographic area where overwintering might be successful. Such risk-averse forecasts may be useful for biosecurity efforts.
IntroductionBeekeeping in rural regions of Peru presents considerable heterogeneity in technification and productivity, which may limit its development. This study aimed to characterize beekeeping systems and identify producer typologies associated with productive characteristics in the district of Tamburco, Apurímac, Peru.MethodsA quantitative, descriptive, cross-sectional study was conducted using a census sample of 34 active beekeepers. Data were collected through a structured and validated questionnaire. Data analysis included descriptive statistics, exploratory hierarchical cluster analysis, association tests, correlation analysis, and complementary multivariate analyses based on FAMD and HCPC.ResultsThe multivariate analyses identified distinct beekeeper typologies that differed in production scale, educational level, technification, and access to training. Significant associations were observed between cluster structure and several productive and sociodemographic variables, while beekeeping experience was positively associated with production scale.DiscussionOverall, the identified typologies were primarily associated with differences in production scale, technological level, and access to training. These findings may help guide extension and training strategies for beekeepers in Tamburco. Given the relatively small sample size and the cross-sectional design, the findings should be interpreted as exploratory and require validation in larger and geographically diverse populations.
Honey bee colony losses are an increasing global concern, yet nationwide pathogen-resolved surveillance data integrating molecular diagnostics with field-based mite monitoring remain limited. This study presents the first national active surveillance program for honey bee diseases in South Korea, combining standardized field inspections with RT-qPCR-based pathogen detection. A total of 1,715 samples were collected from 107 apiaries during three seasonal surveillance rounds in 2025: Round 1 (post-overwintering; March–May), Round 2 (post-honey-harvest; July–August), and Round 3 (pre-overwintering; October–November). Both Apis mellifera and Apis cerana colonies were included in the analysis. Fourteen target honey bee pathogens were screened using RT-qPCR assays (Ct ≤ 35), and Varroa destructor infestation status in A. mellifera apiaries was evaluated through brood inspection or powdered-sugar roll testing depending on seasonal field conditions. DWV prevalence increased progressively from 70.1% in Round 1 (post-overwintering) to 90.4% in Round 2 (post-honey-harvest) and 97.0% in Round 3 (pre-overwintering), coinciding with increasing Varroa detection in A. mellifera apiaries (28.1%, 85.4%, and 87.9%, respectively). Israeli acute paralysis virus (IAPV) also increased progressively across the three surveillance rounds (18.3%, 40.1%, and 47.7%). In contrast, black queen cell virus (BQCV), sacbrood virus (SBV), Vairimorpha spp. (formerly Nosema spp.), and chalkbrood showed declining trends toward autumn. Co-infection patterns shifted seasonally. Although the mean number of detected pathogens per sample decreased from 2.76 to 2.02, co-detection of DWV and IAPV increased markedly from 15.3% to 47.0%. In A. cerana apiaries, SBV remained highly prevalent throughout the surveillance rounds. These findings indicate that the late-season expansion of the Varroa–DWV–IAPV complex represents a major epidemiological feature of Korean apiculture and coincides with the critical period of winter-bee production. This nationwide surveillance framework provides an evidence-based foundation for seasonal honey bee disease management and optimized timing of Varroa control interventions in temperate apicultural systems.
RNA interference (RNAi) has emerged as a promising strategy for insect pest control, yet its application in lepidopterans is hindered by insensitivity to double-stranded RNA (dsRNA). Here, we investigate the piwi-interacting RNA (piRNA) pathway as an alternative, mechanistically distinct gene silencing system in Spodoptera frugiperda, Plutella xylostella, and Cydia pomonella. Small RNA profiling revealed that piRNAs (26–30nt) are the predominant class across all species, marked by strong ping-pong signatures and extensive genomic distribution. Notably, abundant somatic expression, including in gut tissues, indicates that ubiquitous, endogenous piRNA populations may be able to initiate processing of exogenous RNA substrates. To test this possibility, synthetic piRNA-trigger constructs were found to induce gene silencing in Sf9 cells. Further, this approach to gene silencing offers opportunities for nucleic acid engineering, which we demonstrate by incorporating G-quadruplex structures that can enhance RNA stability. Together, these findings establish piRNAs as the dominant small RNA pathway in these lepidopterans and provide a mechanistic and design framework for next-generation RNA-based biopesticides.
Introduction:The consumption of edible insects (EIs) as food has been a long tradition for some populations worldwide. They are sold in markets found in the rural and urban areas of Imo State, Nigeria. Data on the economic value, marketing, and utilization of EIs are limited, particularly in Southeast Nigeria where the study was conducted. This study is necessary for the provision of data on effective marketing and utilization of EIs in order to determine their sustainability in relation to the current market system. Methods:This study investigated the marketability of some EIs as a means of improving rural livelihood in Imo State, Nigeria. Data were collected using structured and validated questionnaires administered interpersonally to 60 respondents/EI marketers in the selected markets. Descriptive and inferential methods were employed in data analysis. Results:The study revealed that four species of EIs were sold: Macrotermes species (35.0%), Gonimbrasia belina (30.0%), Rhynchophorus species (25.0%), and Anaphae species (10.0%), with more than one species sold by marketers (43.8%). EIs were primarily obtained from the wild (68.3%) and were sold mainly for consumption (91.7%); marketing is dominated by women (91.7%). The supply of insects is irregular and seasonality is a major constraint (51.7%). Macrotermes species (termites) had the highest significant mean quality per marketer (p < 0.05). There was a significant difference in mean quality per marketer of G. belina, Rhynchophorus, and Anaphae species (p < 0.05). Discussion:The study concluded that EI marketing is a promising venture that needs attention. Policy intervention that will facilitate mass production/domestication of EIs and modern technical skills on harvesting is recommended.
IntroductionBanana bunchy top disease (BBTD), caused by the banana bunchy top virus (BBTV), poses a growing threat to banana production in Africa and is spread locally by the banana aphid, Pentalonia nigronervosa. Effective BBTD management requires better understanding of aphid habitat, dispersal, response to farm operations, and practical aphid vector and disease control options.MethodsThis study integrated field assessments, field experiments, and a laboratory trial in Uganda to characterize aphid ecology and refine integrated pest and disease management.ResultsAphids were highly cryptic, occurring mainly between leaf sheaths and on lower and middle pseudostem sections. Though winged aphids were mostly associated with larger aphid colonies they occurred across all colony sizes, indicating that even small colonies may contribute to BBTV spread. Sticky traps showed winged aphids to actively disperse throughout fields, with flights often extending beyond adjacent mats. Mechanical disturbance, simulating farm activities such as de-trashing and roguing, strongly increased movement of both winged and wingless aphids and occasionally flight, which could potentially increase BBTV transmission risk in case aphids are viruliferous. Aphids persisted on cut and disposed pseudostem pieces, especially under shaded conditions, but not on corms. In laboratory assays, aphids preferred leaf tissue, although field colonies were mainly pseudostem-associated. Spraying with a 1% laundry soap solution rapidly reduced both winged and wingless aphid populations within 30 minutes to two hours, with populations remaining low for up to two weeks on de-trashed plants.DiscussionThese findings show that BBTD management should target hidden pseudostem colonies, minimize aphid dispersal during plant disturbance, apply soap spray before or immediately after de-trashing infected plants or infected mat removal, and rapidly bury infected pseudostems and leaves while drying corm pieces separately. The study provides practical refinements for smallholder-oriented BBTD integrated management.
This study evaluates two nanocarrier systems, clay-based magnesium-aluminum layered double hydroxide (MgAl-LDH) and the cell-penetrating peptide PepFect14 (PF14), for delivering double-stranded RNA (dsRNA) targeting the αCOP gene in Brassicogethes aeneus. Both carriers successfully formed stable complexes and protected dsRNA from degradation under simulated gut conditions. PF14 produced small, uniform nanoparticles (<120 nm), whereas MgAl-LDH generated substantially larger particles (~416 nm), potentially limiting cellular uptake. Feeding assays revealed that naked dsRNA caused high mortality (93% by day 12), while PF14-complexed dsRNA induced substantial mortality (70%) with a delayed onset. In contrast, MgAl-LDH-complexed dsRNA achieved only 22% mortality despite providing strong protection against degradation. mRNA expression analysis at day 3 and 6 showed moderate, statistically insignificant early αCOP downregulation, consistent with delayed intracellular processing. At higher concentrations (600 ng/µL), naked dsRNA strongly suppressed αCOP transcripts (100%), whereas MgAl-LDH complexes produced only modest knockdown (63.6%). Overall, these findings suggest that while MgAl-LDH offer robust dsRNA stabilization, its delivery efficiency may be constrained by particle size and gut physiology. PF14 demonstrates promise as a carrier enabling sustained delivery with delayed onset, whereas MgAl-LDH requires higher dsRNA doses to achieve comparable effects. Carrier selection should balance dsRNA stability with timely release and account for species-specific gut barriers to optimize RNAi-based pest control strategies.
Neuropeptides are evolutionarily ancient signaling molecules that act through diverse G protein-coupled receptor (GPCR) pathways to coordinate essential physiological and behavioral processes in insects. Despite their broad functional importance, the specific roles, receptor interactions, and regulatory mechanisms of individual neuropeptides remain underexplored, limiting our understanding of how these signaling systems operate across developmental and physiological contexts. In this study, we characterized transcript expression of five functionally diverse neuropeptides and one GPCR across Leptinotarsa decemlineata (Colorado potato beetle) tissues and life stages and subsequently knocked down the glycoprotein hormone subunit alpha 2 (GPA2) and its cognate receptor with glycoprotein hormone subunit beta 5 (GPB5), leucine-rich repeat-containing GPCR 1 (LGR1), for phenotypic investigation. Expression analyses revealed broad and tissue-specific variation among neuropeptides across developmental stages. No significant patterns in mortality, larval weight, or defoliation were detected following dsRNA-mediated knockdown of GPA2 or LGR1 via injection, though in the feeding trial third instar larvae given LGR1 dsRNA weighed significantly more and showed reduced defoliation rates compared the control. State-transition modeling identified significant effects of GPA2 and LGR1 knockdown on developmental timing, specifically during pupal-adult and larval-pupal-adult transitions, respectively, with time in life stage emerging as a key predictor of overall survival. These findings suggest a potential hormonal role for the GPA2-LGR1 signaling system in regulating developmental progression and stabilizing life-stage transitions in L. decemlineata.
IntroductionApple production in Kazakhstan is highly vulnerable to damage caused by fruit-infesting tortricid pests, particularly codling moths (Lepidoptera: Tortricidae). Accurate and rapid species identification is essential for effective pest management and phytosanitary monitoring in apple orchards.MethodsField surveys were conducted in apple orchards of southern and southeastern Kazakhstan to investigate the codling moth complex. Collected specimens were identified morphologically and confirmed by sequencing a mitochondrial cytochrome c oxidase subunit I (COI) region. Comparative COI sequence analysis was used to develop a multiplex quantitative polymerase chain reaction assay employing Minor Groove Binder hydrolysis probes for simultaneous detection and differentiation of Cydia pomonella and Grapholita molesta.ResultsField surveys identified C. pomonella and G. molesta, with C. pomonella accounting for 83.3% of collected specimens. COI sequencing enabled reliable discrimination between the two species. The multiplex assay demonstrated high specificity, produced no false-positive amplification in non-target species, and achieved 100% detection down to 5 pg DNA per reaction.DiscussionThe developed COI-based multiplex quantitative polymerase chain reaction assay provides a rapid, sensitive, and reliable tool for simultaneous identification of C. pomonella and G. molesta. It is suitable for phytosanitary monitoring, early detection, and management of quarantine-relevant tortricid pests in Kazakhstan.
The honey bee gut microbiota plays a crucial role in host nutrition, immunity, and colony health, yet the relative influence of seasonal and colony-specific factors on its long-term dynamics remains incompletely understood. This study investigated temporal variation in the gut bacterial community of three Apis mellifera ligustica colonies maintained in the same apiary and monitored over two consecutive years (2022–2023). Worker bees were sampled during eight seasonal periods, and gut microbiota composition was characterized using 16S rRNA gene amplicon sequencing. Across all sampling periods, the microbiome was consistently dominated by the characteristic honey bee-associated genera Gilliamella, Snodgrassella, Bartonella, Frischella, Commensalibacter, and Lactobacillus, indicating the persistence of a conserved core bacterial community. Seasonal variation was primarily associated with changes in the relative abundance of dominant taxa rather than with major changes in community composition. In particular, Gilliamella apicola and Snodgrassella alvi exhibited complementary seasonal patterns, with Gilliamella reaching its highest abundance during autumn, particularly in autumn 2023, whereas Snodgrassella predominated during spring and winter. Alpha-diversity metrics (Observed OTUs, Chao1, Shannon, and Simpson indices) showed limited seasonal variation, whereas beta-diversity analyses detected significant differences in community composition among seasons. Principal Coordinates Analysis and PERMANOVA identified season as the factor most strongly associated with microbiome variation, while colony identity did not significantly influence bacterial community composition under the standardized experimental conditions adopted in this study. Overall, these findings show that the gut microbiome of A. mellifera ligustica maintains a conserved core bacterial community while exhibiting reproducible seasonal variation in the relative abundance of its dominant members. This study provides a longitudinal baseline for future investigations aimed at understanding the ecological mechanisms underlying seasonal microbiome dynamics and their relationship with honey bee biology and environmental change.
Background:Food crop infestations caused by the insect pest Locustana pardalina threaten global food security, particularly in developing regions. This pest can devastate entire crops, exacerbating malnutrition and economic instability. Traditionally, L. pardalina outbreaks have been managed using synthetic chemical insecticides, which, while effective, pose significant environmental risks by harming nontarget species and contaminating ecosystems. Given these concerns, there is an increasing demand for environmentally friendly alternatives. Medicinal plant extracts, known for their biodegradability, cost-effectiveness, and active metabolites, offer promising solutions. Method:A systematic literature search was conducted across Google Scholar, PubMed, ScienceDirect, and SpringerLink using keywords related to Locustana pardalina, food security, outbreaks, botanical insecticides, metabolites, and pest management. The search (1988-2024) conducted used predefined eligibility criteria and identified 120 relevant studies on the biological control of various insects spanning approximately five to eight crop systems. Results:These plant-based insecticides have demonstrated the ability to repel various insect pests, including L. pardalina, and could provide an effective method for controlling locust swarms while mitigating the harmful effects of chemical pesticides. This review discusses the potential of medicinal plant extracts as sustainable alternatives for managing L. pardalina infestations, highlighting the active compounds that show promise in pest control and their benefits over conventional chemical insecticides. Conclusion:In conclusion, medicinal plant extracts represent a sustainable and environmentally friendly alternative to synthetic insecticides for managing Locustana pardalina infestations. Their biodegradability, cost-effectiveness, and bioactive metabolites offer effective pest control while minimising ecological harm, highlighting their potential to support food security and reduce the negative impacts associated with chemical pesticides.
