Tribolium Castaneum Research Articles

Traits Underlying Experimentally Evolved Dispersal Behavior in Tribolium castaneum

AbstractDispersal is an important behavior in many animals, with profound effects on individual fitness and the evolutionary trajectories of populations. This is especially true within taxa with particular life-history strategies, for example those that exploit ephemeral habitat. Further, dispersal is commonly seen to be part of behavioral syndromes - suites of traits that covary across behavioral contexts. The red flour beetle, Tribolium castaneum (Coleoptera, Tenebrionidae), is a major post-harvest crop pest responsible for large losses through the infestation of stored grain. In this system dispersal is known to have a strong genetic basis and differential artificial selection on dispersal traits produces strong phenotypic divergence. However, it is unknown which traits are able to rapidly evolve to produce these results, or which behavioral components underlie differences in dispersal. Using replicate lines of T. castaneum previously selected for divergent dispersal behavior, we test for correlated activity and movement patterns, morphology and substrate surface use. We find robustly repeatable associations between the dispersal phenotype and higher activity, straighter paths, larger body size (but not relative leg length) and increased tendency to remain at the surface of fodder. Together our results suggest that dispersal is part of a syndrome of traits in T. castaneum, and must be treated as such when considering the evolution of dispersal in this system, and in attempting to predict and control its spread.

Density dependent survival drives variation in density dependent population growth of an insect pest

Several ecological processes – from population dynamics to species co‐existence – are driven by density dependence (DD) in population growth rate. Thus, to predict and manage ecological outcomes, we need a deep understanding of which factors and demographic traits drive variation in DD. In the insect pest Tribolium castaneum, we found large variation in DD across habitats but not across source populations. We modeled DD in population growth as the product of DD in fecundity and survival, experimentally estimating each parameter. Across habitats, survival parameters varied more than fecundity, including in simulations with varying parameter values. Thus, DD in survival drives variation in density‐dependent population growth. Hence, under strong density‐dependent selection, we expect evolutionary change in density‐dependent survival, provided sufficient genetic variance. Our general framework combining detailed experiments and simulations with a simple model can be used for other species to better understand the causes and consequences of density dependent population growth.

Efficient CRISPR‐mediated genome editing can be initiated by embryonic injection but not by ovarian delivery in the beetle Tribolium castaneum

Efficient CRISPR‐mediated genome editing can be initiated by embryonic injection but not by ovarian delivery in the beetle <i>Tribolium castaneum</i>

Knockdown of TcGluCl leads to the premature pupation of Tribolium castaneum larvae possibly by influencing the calcium-mediating hormone homeostasis

The glutamate-gated chloride channels (GluCls) are widely existed in the neural and nonneural tissues of invertebrate. In addition to play important roles in signal transduction, the GluCls also showed multiple physiological functions in insects such as participate in the juvenile hormone synthesis. In the present study, the potential roles of TcGluCl in growth and development of the red flour beetle Tribolium castaneum were explored. Knockdown of TcGluCl showed no effects on the survivability, weight growth, final pupation rate, eclosion and fecundity of T. castaneum, whereas resulted in the significant premature pupation of larvae. Inhibition of TcGluCl expression significantly changed the levels of juvenile hormone and ecdysone as well as the expressions of hormone biosynthetic genes. The increased ecdysone level and decreased juvenile hormone level were observed at the late stage of dsGluCl-treated larvae. Knockdown of TcGluCl significantly reduced the expressions of TcSTIM1 and TcOrai1, which were the primary proteins in store-operated calcium entry (SOCE) mediated Ca2+ influx mechanism. Whilst the L-glutamic acid treatment led to the increased TcOrai1 expression in T. castaneum. These findings suggested that knockdown of TcGluCl increased the ecdysone level and contributed to the premature pupation of larvae, which might be due to the reduced Ca2+ influx caused by the decreased expressions of TcSTIM1 and TcOrai1. These studies provide novel insights on the function of GluCls in insects.

Nutrition affects larval survival and the development of morphological traits in male and female flour beetles, but genital size and shape remains canalised.

The caloric content and macronutrient ratio of diet consumed is a major source of phenotypic variation in most animal populations. While these nutritional effects have been well-documented for a variety of life-history and morphological traits, the effects of nutrition on male genitals are poorly understood but genitals are thought to be more canalised than general morphology and hence less susceptible to variation in nutrition. Even less is known about the effects of nutrition on female genital form, which to our knowledge, have never been investigated. Here we tested for effects of juvenile dietary macronutrients (protein and carbohydrate) on larval survival, adult morphology, including genital size and shape in male and female flour beetles (Tribolium castaneum). We found there was nutritionally induced plasticity in larval survival and morphology, although the latter effect was variable, with body size being most responsive to dietary macronutrients and genital size and shape being least responsive. Functionally equivalent morphological traits in the sexes responded similarly to nutrition. Previously, we showed that the genitalia of male and female T. castaneum are subject to strong stabilising sexual selection, and our current findings suggest that developmental mechanisms reduce the nutritional sensitivity of male and female genitals, possibly to ensure matching during mating.

Experimental evolution reveals that males evolving within warmer thermal regimes improve reproductive performance under heatwave conditions in a model insect.

Climate change is increasing mean temperatures, and intensifying heatwaves. Natural populations may respond to stress through shorter-term acclimation via plasticity and/or longer-term inter-generational evolution. However, if the pace and/or extent of thermal change is too great, local extinctions occur; one potential cause in ectotherms is identified to be the heat-liability of male reproductive biology. Recent data from several species, including the beetle Tribolium castaneum, confirmed that male reproductive biology is vulnerable to heatwaves, which may constrain populations. However, such reproductive-damage may be overestimated, if there is potential to adapt to elevated mean temperatures associated with climate change via evolution and/or acclimation. Here, we tested this to evaluate whether pre-exposures could improve heatwave tolerance (adaptation or acclimation), by experimentally evolving T. castaneum populations to divergent thermal regimes (30°C versus 38°C). Findings across assays revealed that relative to 30°C-regime males, males from the 38°C regime, maintained constantly at 8°C warmer for 25 generations, displayed an increase; i) in post heatwave (42°C) reproductive fitness by 55%, ii) survival by 33% and iii) 32% larger testes volumes. Unexpectedly, in the acclimation assay, warm-adapted males' post-heatwave survival and reproduction were best if they experienced cool developmental acclimation beforehand, suggesting a cost to adapting to 38°C. These results help progress knowledge of the potential for survival and reproduction to adapt to climate change; trait specific adaptation to divergent thermal regimes can occur over relatively few generations, but this capacity depended on the interaction of evolutionary and thermal acclimatory processes.

Isolation of Bioactive Fumigants from Different Varieties of Cassava (Manihot esculenta Crantz) and their Toxicity on Tribolium castaneum Herbst and Rhyzopertha dominica Fabricius

Isolation of Bioactive Fumigants from Different Varieties of Cassava (Manihot esculenta Crantz) and their Toxicity on Tribolium castaneum Herbst and Rhyzopertha dominica Fabricius

Modeling long-term, stage-structured dynamics of Tribolium castaneum (Coleoptera: Tenebrionidae) at food facilities with and without two types of long-lasting insecticide-incorporated netting.

The red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), is a cosmopolitan and destructive external-infesting pest at many food facilities. The use of deltamethrin- and α-cypermethrin-incorporated long-lasting insecticide-incorporated netting (LLIN) has shown incredible promise for the management of stored product insects. However, it is unknown how LLIN deployed within food facilities may affect the long-term population dynamics of T. castaneum compared to populations where no LLIN is present. Exposure to LLIN has been shown to affect mortality in the current generation and decrease progeny production in the subsequent generation. Thus, we modeled the long-term population dynamics of T. castaneum at food facilities over 15 generations by incorporating realistic estimates for mortality and progeny reduction after contact with LLIN compared to baseline growth by the species. We parameterized the model with estimates from the literature and used a four-stage structured population (eggs, larvae, pupae, and adults). The model was implemented using the package popbio in R. Our models suggest that deploying LLIN led to significant population reductions based on the estimates of mortality and progeny reduction from prior work, whereas the baseline model exhibited exponential population growth. In addition, there were differences in the frequencies of each life stage under each scenario modeled. As a result, it appears deploying LLIN may contribute to the local extirpation of T. castaneum within as few as 15 generations. Our work contributes to a growing literature about the effectiveness of incorporating LLIN into existing pest management programs for managing stored product insects in food facilities.

Repellency, Fumigant Toxicity, Antifeedent and Residual Activities of Coridothymus capitatus and Its Main Component Carvacrol against Red Flour Beetle.

Tribolium castaneum is a challenging pest of stored products, causing significant economic losses. The present study explored the efficacy of Coridothymus capitatus essential oil and its primary constituent, carvacrol, as eco-friendly alternatives for managing this pest. To evaluate their insecticidal potential, repellency, fumigant toxicity, and antifeedant properties, progeny inhibition assays were performed. Carvacrol exhibited superior repellency compared to the essential oil, achieving a 92% repellency rate at 2 mg/cm2. Both compounds demonstrated significant fumigant toxicity against T. castaneum, with LC50 values of 168.47 and 106.5 μL/L for the essential oil and carvacrol, respectively, after 24 h. Carvacrol also outperformed the essential oil in antifeedant activity, inducing an 80.7% feeding deterrence at 1.17 mg/g. Moreover, both treatments effectively suppressed the development of the pest's progeny. These results collectively underscore the potent insecticidal properties of C. capitatus essential oil and carvacrol, particularly carvacrol, as promising candidates for the sustainable management of T. castaneum in stored product protection.

Carboxylesterase α-EST5 is required for the red flour beetle to metabolize mugwort essential oil

The red flour beetle (Tribolium castaneum) is a global agricultural grain storage pest that seriously jeopardizes the food safety of wheat, corn, and other grains. Carboxylesterases belong to a family of multifunctional enzymes that can participate in the metabolism of toxic substances ingested by T. castaneum, conferring drug resistance. In this study we have cloned from T. castaneum a gene encoding carboxylesterase, α-EST5, which is expressed mainly in T. castaneum late larvae and late adults. Tissue expression pattern analysis showed that α-EST5 was mainly expressed in the larval gut and adult head. Moreover, α-EST5 was able to be induced to be expressed by mugwort essential oils and significantly increased the enzymatic activity of carboxylesterase in T. castaneum after mugwort essential oil treatment. The sensitivity of T. castaneum to mugwort essential oils was significantly increased after knockdown the expression of α-EST5 using RNAi (RNA interference) technology. Through our study, we demonstrated that the carboxylesterase α-EST5 plays an important role in the detoxification of mugwort essential oils by T. castaneum. This study results provide a reference for understanding the detoxification mechanism of T. castaneum to external toxic substances.

In Vitro Evaluation of Native &lt;i&gt;Bacillus thuringiensis&lt;/i&gt; (Berliner) Isolates against Different Insects

Evaluation of native strains of Bacillus thuringiensis (Berliner) isolates against different orders of insects and characterization of responsible Cry genes was carried out. Among the native isolates, BGC-1 showed the least LC50 value of 5.24 μg/ ml and was comparable to the reference strain HD1 (2.89 μg/ ml) against tobacco caterpillar Spodoptera litura (Fabricius). The isolate RCM-1 recorded least LC50 value of 4.69 μg/ ml against the housefly Musca domestica Linnaeus. One isolate viz., GHB-1 was found to be potential against both the larvae of S. litura and M. domestica. The isolate RCM-2 registered the least LC50 value of 8.21 μg/ ml against grubs of red flour beetle Tribolium castaneum (Herbst). Several isolates had more than one Cry gene in them. Among them, 12 isolates (63.15%) were found to contain Cry3 genes, 11 isolates harbour Cry4 genes, five isolates had Cry1, Cry2 and Cry11 genes each and four isolates contain Cry7 genes each. Native isolate, GHB-1 had Cry1, Cry3, Cry4 and Cry11 genes.

Toxicity and Repellent Effect of Three Plant Based Essential Oils against the Red Flour Beetle &lt;i&gt;Tribolium castaneum&lt;/i&gt; (Herbst)

In this study jasmine (Jasminum officinale), lemon grass (Cymbopogon citratus), and sandalwood (Santalum album) essential oils were investigated for their efficacy against the red flour beetle Tribolium castaneum. Sandalwood oil consistently showed the highest contact toxicity, with LD50 values decreasing over time. Jasmine and lemon grass oils ranked lower but still exhibited toxicity. The order of contact toxicity was sandalwood &gt; jasmine &gt; lemon grass at each exposure period. Fumigation treatments revealed sandalwood’s initial superiority, though jasmine surpassed it at 48 hr. The order of fumigation toxicity shifted to jasmine &gt; sandalwood &gt; lemon grass at this extended exposure time. Repellency tests showed dose and time-dependent responses for both larval and adult stages, with occasional fluctuations. Sandalwood’s significance differed for adult beetles between time intervals, and jasmine exhibited insignificance in its efficacy for larvae at certain intervals. This study provides insights into the potential of these essential oils as alternatives for pest control.

RNA interference-mediated silencing of ctl13 inhibits innate immunity and development in stored pest Tribolium castaneum

C-type lectins (CTLs) play a pivotal role in the regulation of insect immunity and growth, making them potential molecular targets for RNA interference (RNAi)-mediated pest control. Although multiple CTLs have been identified in the genomes of various insects, their specific functions and underlying molecular mechanisms remain unclear. In the present study, a novel CTL, Tcctl13 with a single CRD, was identified in Tribolium castaneum. Tcctl13 is expressed in diverse immune-related tissues and developmental stages, with a notable increase in its expression upon exposure to lipopolysaccharides (LPS) and peptidoglycan (PGN). Molecular docking and enzyme-linked immunosorbent assay (ELISA) analyses revealed that TcCTL13 possesses the ability interacted with LPS and PGN. The binding and agglutinating activities of recombinant TcCTL13 (rTcCTL13) were demonstrated against both gram-negative and positive bacteria. After using RNAi to silence Tcctl13, the expression of the eight antimicrobial peptide (AMP) genes was significantly reduced. In addition, knocking down Tcctl13 during the early larval or pupal stage hindered, the normal metamorphosis process in T. castaneum, ultimately leading to the demise of all beetles. Further research showed that Tcctl13 and nine AMPs were significantly downregulation after 20-Hydroxyecdysone (20E) injection. Instead, the up-regulation of Tcctl13 and six AMPs was observed following interference with the 20E receptor (ecdysone receptor, EcR), indicating that the function of Tcctl13 is regulated by 20E in T. castaneum. Collectively, these findings suggest that Tcctl13 plays a role in the regulation of innate immunity and development in T. castaneum, offering a promising molecular target for managing insect pests using RNAi-based approaches.

Studies on the Response of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) to Multicolour Light System

Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) commonly known as rust red flour beetle was a universal pest of stored produce with an extensive feeding diversity. The management of T. castaneum in field conditions was through fumigation and spraying of suitable insecticides. Even though many plant products were found effective against pests in invitro conditions, they could not be mixed with the stored produce for fear of losing the table quality. Physical forces seemed to be good alternatives to chemical treatment as far as stored grain pests were concerned. The use of gamma and UV radiations, microwaves and ultrasonic sound waves against T. castaneum and other stored grain pests had been tried by many authors. The objectives of this study were to check the adult beetles and final instar grubs of T. castaneum to different colours of light and it showedpreference for clear tungsten (white) light, while they were averse to red coloured light. This trend was explicit even after the entrainment of beetles to red colour light for further generations. Based on these results it could be presumed that T. castaneum adults preferred to move away from red coloured light. By changing the colour of bright light to dark lighting in warehouses, it could be possible to manage T. castaneum in small measure. Total eradication of stored product pests like T. castaneum was at least possibility and the selection of a proper management system could help to solve the stored produce pest problem to a greater magnitude.

A sex-specific homologue of waprin is essential for embryonic development in the red flour beetle, Tribolium castaneum.

Waprin, a WAP (Whey acidic protein) domain-containing extracellular secretory protein, is widely known for its antibacterial properties. In this study, a waprin homologue (Tc_wapF) expressing in a female-specific manner was identified in Tribolium castaneum, through the analysis of sex-specific transcriptomes. Developmental- and tissue-specific profiling revealed the widespread expression of Tc_wapF in adult female tissues, particularly in the ovary, gut and fatbody. This female-specific expression of Tc_wapF is not regulated by the classical sex-determination cascade of T. castaneum, as we fail to get any attenuation in Tc_wapF transcript levels in Tcdsx and Tctra (key players of sex determination cascade of T. castaneum) knockdown females. RNA interference-mediated knockdown of Tc_wapF in females led to the non-hatching of eggs laid by these females, suggesting the crucial role of Tc_wapF in the embryonic development in T. castaneum. This is the first report on the identification of a sex-specific waprin homologue in an insect and its involvement in embryonic development. Future investigations on the functional conservation of insect waprins and their mechanistic role in embryonic development can be exploited for improving pest management strategies.

The feeding preference of red flour beetle Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) on wheat flour stored at varied temperatures: The perspective of volatile components

As a common pest in wheat flour, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) is responsible for economic losses and potential risks to food safety. This study aimed to investigate the feeding preferences of T. castaneum on wheat flour stored at different temperatures, analyzed the key flavor compounds influencing its feeding preferences, and laid down the foundation for the development of new T. castaneum attractants and repellents. According to results, in the early storage period, T. castaneum showed a higher preference for feeding on wheat flour stored at higher temperatures, but this trend decreased over the storage time with a preference shift towards the flour preserved at lower temperatures. The findings from GC-MS revealed changes in the types and concentrations of volatile components in wheat flour during storage. PCA and correlation analysis suggested that compounds such as 2-methyldodecane, 3-methyl-3-ethylheptane, benzaldehyde, decanal, and n-hexanol exhibited potential attractiveness to T. castaneum. Conversely, 2-methyloctane, 2,5-dimethyldodecane, and dodecane were identified as potential repellents. At different storage temperatures, benzaldehyde and 2-methyloctane displayed consistent attractant and repellent effects, indicating their potential as effective components for attractance and repellence. Overall, our work shed the light regarding the feeding preference of T. castaneum on wheat flour stored at different temperatures from the perspective of volatile component variations, and provided a theoretical basis for the development of novel attractants and repellents.

Challenges in Assessing Repellency via the Behavioral Response by the Global Pest Tribolium castaneum to Protect Stored Grains.

Food security is an increasingly pressing global issue, and by 2050, food production will not be sufficient to feed the growing population. Part of global food insecurity can be attributed to post-harvest losses, including quantity and quality losses caused by stored-product pests like insects. It is thus timely to find management strategies to mitigate these losses and counteract food insecurity. The red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), a global stored-product pest with a wide range of food sources, was used in this study to assess repellency to a selection of essential oils. Multiple behaviorally relevant methods were used to determine the efficacy of the essential oils in assays to pinpoint the most promising repellents. Experiments were used to assess individual and group behaviors with or without airflow and examined the behavioral variation in distance moved and the time spent away from the oil. It was found that exposure to essential oils and conditions of experimentation considerably influenced T. castaneum's behavioral response, but a clear candidate for repellency could not be chosen based on the collected data. Follow-up research is needed to pinpoint repellents for integrated pest management practices to protect grains from stored-product pests and to justify their use in and around commodities.

Antifeedant and growth inhibitory effects of extracts from Ajania potaninii and A. fruticulosa against Tribolium castaneum (Coleoptera: Tenebrionidae)

This study aimed to evaluate the nutritional and antifeedant indices of Tribolium castaneum in response to the compounds and main chemical constituents ofAjania potaninii and A. fruticulosa. These mainly includes essential oils and their major ingredients (eucalyptol, camphor, myrtenol, verbenol, and borneol), as well as nonvolatile compounds. The A. fruticulosa and A. potaninii EOs exhibited clear antifeedant activity against T. castaneum adults at a 2 mg/L concentration, with FDI of 50% and 60%, respectively. Among the nonvolatile compounds, sesamin and eriodictyol exhibit strong antifeedant activities against T. castaneum adults at the same concentration, with an FDI of 90.57% and 88.68% after 72 h, respectively. For larvae, A. fruticulosa and A. potaninii EOs also resulted in antifeedant activity (respective FDI: 57.08% and 55.63%). In addition, sesamin and eriodictyol had exceptionally potent antifeedant activity against T. castaneum larvae, with FDI values 90.71% and 96.17%, respectively. In nutritional studies, the reductions in relative food consumption rate (RCR) of T. castaneum were more pronounced for 1 mg/L and 2 mg/L of either eucalyptol or sesamin. However, at greater concentrations, the conversion efficiency of eucalyptol and sesamin to ingested food in T. castaneum was significantly improved. Eucalyptol and sesamin showed a dose-dependent FDI for this insect. Considering the significant anti-insect activities, the EOs, main compounds, and non-volatile compounds of both A. potaninii and A. fruticulosa might be useful in devising integrated pest strategies, providing a promising perspective for the comprehensive use of natural plant resources from the Ajania genus.

Detection Limits of Insect Fragments in Spiked Whole Wheat Flour Using Multiplex Polymerase Chain Reaction (PCR)

The need for a sensitive molecular method to detect specific species of insect contaminants in food products remains a significant challenge in the food industry. This study evaluated the detection limit of a multiplex end-point PCR assay for detecting insects in food. The assay amplifies two fragments of the cytochrome oxidase subunit I gene (COI-Fa and COI-Fb) and one fragment of the protein-coding wingless (wg) gene found in insects. Five insect species, comprising three vectors of foodborne pathogens (the housefly, Musca domestica, the American cockroach, Periplaneta americana, and the pharaoh ant, Monomorium pharaonis) and two storage insect pests (the red flour beetle, Tribolium castaneum and the Indian meal moth, Plodia interpunctella), were spiked separately and in combination at levels of 1, 0.1, 0.01, and 0.001% in whole wheat flour. At spike levels greater than 0.01%, amplicon bands of expected sizes were seen in 100% of samples containing fragments from distinct insect species. At least 25% of spiked samples at the lowest spike level had amplicon bands, except for samples spiked with M. domestica. Results showed an 18.9% probability (with 11.3% and 30% lower and upper confidence limits, respectively) of detecting insect fragments at the lowest spike level (0.001%, corresponding to 3–22 fragments), which is far below the FDA’s regulatory level of less than 75 fragments per 50 g of wheat flour. The intensity of amplicon bands in the gel images was higher at higher spike levels. However, this method is not quantitative enough to extrapolate the intensity of the amplicon bands to the number of insect fragments present in a sample. This multiplex assay was also evaluated in a variety of market food samples derived from plants and animals, showing its potential use in various food types. Overall, the sensitivity and specificity of this molecular approach suggest that it could be used in the future as a screening tool for detecting insect contaminants in food.

Species Composition and Diversity of Stored Grain Pests in Fortified Rice Collected from Different Districts of Telangana State, India

A study was conducted to measure the species composition and diversity of various stored grain insect pests in fortified rice collected from different districts of Telangana,India during 2023-2024. Theresults indicated that a total of six species of stored grain insects belonging to three orders viz., Coleoptera (3), Lepidoptera (2) and Psocoptera (1) were recorded in fortified rice. The six insect species were rice weevil, Sitophilus oryzae, Saw toothed beetle, Oryzaephilus surinamensis, redflour beetle, Tribolium castaneum, Angoumois grain moth, Sitotroga cerealella, rice moth, Corcyra cephalonica and pscocids. Among all these insects highest population of O. surinamensis was recorded in four districts of Telangana except in Ranga reddy and Nalgonda. The diversity indices indicated that Shannon-weiner diversity index (H) was high in Warangal district samples(H=1.426) while the Jagtial population recorded highest Margalef’s species richness index (R=1.27). The highest Pielou’s evenness index was reported from Nalgonda district (e=75), and Simpson diversity index of stored grain pests was found to be maximum (0.718) in Warangal district rice samples. The knowledge on species composition and diversity helps in planning timely management strategies for stored grain pests.