Tribolium Castaneum Research Articles

Beyond adult models: Tribolium castaneum larval timekeeping reveals unexpected robustness and insights into circadian clock.

Circadian rhythms are self-sustained endogenous oscillations that are found in all living organisms. In insects, circadian rhythms control a wide variety of behavioral and physiological processes, including feeding, locomotion, mating, and metabolism. While the role of circadian rhythms in adult insects is well-understood, it is largely unexplored in larvae. This study investigates the potential for larval synchronized activity in the red flour beetle (Tribolium castaneum), a species exhibiting solitary and aggregation phases. We hypothesized that, similar to adults, larvae would exhibit a daily activity pattern governed by an endogenous circadian clock. We further predicted that the transition between the solitary and gregarious phases extends to unique temporal activity patterns. Our results revealed unique timekeeper gene expression in larvae, leading to a distinct daily rhythm characterized by nocturnal activity. Cues indicating on potential cannibalism did not change daily activity peak. However, the absence of these cues significantly reduced the proportion of rhythmic larvae and led to higher variation in peak activity, highlighting the crucial role of social interactions in shaping their rhythmicity. This study sheds light on the evolution and function of larval synchronization in group-living insects, offering novel insights into this complex behavior.

Contact Toxicity and Repellent Effects of Essential Oils from Toddalia asiatica Against Two Stored-Product Insects.

Insect infestations continually endanger stored goods, underscoring the significance of discovering eco-friendly insecticides for pest management. Essential oils (EOs) from different parts of Toddalia asiatica (leaf, fruit and branch) were extracted by hydrodistillation and analyzed by GC-MS. Carvene, p-cymene and muurolene are the principal compounds of T. asiatica leaf (TAL), T. asiatica fruit (TAF) and T. asiatica branch (TAB) EO respectively. Our work aimed to assess the contact toxicity and repellent effects of EOs on two storage pests, Tribolium castaneum and Lasioderma serricorne. All tested EOs exhibited obvious contact toxicity, especially, TAL EO against T. castaneum (33.48 μg/adult) and TAF EO against L. serricorne (16.42 μg/adult). Repellency tests revealed that TAL and TAF EOs, at a concentration of 78.63 nL/cm2, achieved nearing 100 % efficiency against T. castaneum. These results suggest that EOs of T. asiatica could be used as effective botanical insecticides for managing stored-product insects.

Effects of nutrition on recovery, mortality, and mobility of adult Tribolium castaneum after exposure to long-lasting insecticide-incorporated netting.

Long-lasting insecticide-incorporated netting (LLIN) has been used to deliver contact insecticides as an integrated pest management tool for stored product insect pests in food facilities. Although the presence of food is known to improve insect recovery after exposure, it is not clear whether food nutritional quality plays a role. Here, the red flour beetle Tribolium castaneum adults were exposed to two commercially available LLINs, Carifend (active ingredient α-cypermethrin) and D-Terrence (deltamethrin), then transferred to Petri dishes with foods with varying nutritional quality (e.g., 0-100% ratios of flour to non-nutritive cellulose). We investigated the effects of nutrition, LLIN type, and exposure time on post-exposure recovery, mortality, and mobility. After exposure for 2-168 h, the immediate mortality of T. castaneum adults ranged from 0.5% to 91.0% with Carifend and 0% to 75.3% with D-Terrence. Adult recovery and delayed mortality were significantly affected by nutritional quality, LLIN type, exposure time, and recovery time. For both LLINs, adult recovery increased over time, with a trend for higher recovery and lower mortality with increasing nutritional quality and decreasing exposure time. In addition, adult mobility decreased multiple-fold after Carifend or D-Terrence exposure for 30, 60 or 90 min compared to 10 min. This study shows nutrition significantly modulates the efficacy of LLIN against T. castaneum, and thus strengthens the rationale for implementing stringent sanitation protocols for food facility managers. © 2024 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Genome-wide identification and molecular evolution of elongation family of very long chain fatty acids proteins in Cyrtotrachelus buqueti

To reveal the molecular function of elongation family of very long chain fatty acids(ELO) protein in Cyrtotrachelus buqueti, we have identified 15 ELO proteins from C.buqueti genome. 15 CbuELO proteins were located on four chromosomes. Their isoelectric points ranged from 9.22 to 9.68, and they were alkaline. These CbuELO proteins were stable and hydrophobic. CbuELO proteins had transmembrane movement, and had multiple phosphorylation sites. The secondary structure of CbuELO proteins was mainly α-helix. A total of 10 conserved motifs were identified in CbuELO protein family. Phylogenetic analysis showed that molecular evolutionary relationships of ELO protein family between C. buqueti and Tribolium castaneum was the closest. Developmental transcriptome analysis indicated that CbuELO10, CbuELO13 and CbuELO02 genes were key enzyme genes that determine the synthesis of very long chain fatty acids in pupae and eggs, CbuELO6 and CbuELO7 were that in the male, and CbuELO8 and CbuELO11 were that in the larva. Transcriptome analysis under different temperature conditions indicated that CbuELO1, CbuELO5, CbuELO12 and CbuELO14 participated in regulating temperature stress responses. Transcriptome analysis at different feeding times showed CbuELO12 gene expression level in all feeding time periods was significant downregulation. The qRT-PCR experiment verified expression level changes of CbuELO gene family under different temperature and feeding time conditions. Protein-protein interaction analysis showed that 9 CbuELO proteins were related to each other, CbuELO1, CbuELO4 and CbuELO12 had more than one interaction relationship. These results lay a theoretical foundation for further studying its molecular function during growth and development of C. buqueti.

Novel Inhibitor of Glutamate-Cysteine Ligase Catalytic Subunit against Tribolium castaneum: High-Throughout Virtual Screening, Molecular Docking and Dynamics Simulation, and Bioassay.

The enzyme glutamate-cysteine ligase catalytic subunit (Gclc) is a rate-limiting enzyme in the biosynthesis of glutathione that is involved in antioxidant defense, detoxification of xenobiotics, and/or its metabolites and regulates the cell cycle and immune function. Therefore, Gclc presents an appealing target for the development of novel insecticides. In this study, we conducted high-throughput virtual screening from the ZINC20 database and identified three compounds with high binding affinity to the Tribolium castaneum Gclc (TcGclc). Ultimately, we selected ZINC000032992384 due to its superior stability and lowest binding energy, as determined through molecular dynamics simulations. Bioassay results revealed that the IC50 value of ZINC000032992384 was 19.70 μM lower than that of BSO (49.67 μM). Furthermore, the larval mortality in the ZINC000032992384 treated group was 63.8%, significantly higher than that of the controls (29.1% in the dichlorvos group and 6.4% in the acetone group). This study provides novel insights for the development of a Gclc-targeted inhibitor as a potent insecticide based on the interaction between receptors and ligands.

Deciphering a Beetle Clock: Individual and Sex-Dependent Variation in Daily Activity Patterns.

Circadian clocks are inherent to most organisms, including cryptozoic animals that seldom encounter direct light, and regulate their daily activity cycles. A conserved suite of clock genes underpins these rhythms. In this study, we explore the circadian behaviors of the red flour beetle Tribolium castaneum, a significant pest impacting stored grain globally. We report on how daily light and temperature cues synchronize distinct activity patterns in these beetles, characterized by reduced morning activity and increased evening activity, anticipating the respective environmental transitions. Although less robust, rhythmicity in locomotor activity is maintained in constant dark and constant light conditions. Notably, we observed more robust rhythmic behaviors in males than females with individual variation exceeding those previously reported for other insect species. RNA interference targeting the Clock gene weakened locomotor activity rhythms. Our findings demonstrate the existence of a circadian clock and of clock-controlled behaviors in T. castaneum. Furthermore, they highlight substantial individual differences in circadian activity, laying the groundwork for future research on the relevance of individual variation in circadian rhythms in an ecological and evolutionary context.

Impact of Different Food Sources on the Developmental Biology and Host Preferences of the Red Flour Beetle Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae)

An experiment conducted at the seed center laboratory at Tamil Nadu Agricultural University during June 2022 evaluated the host preferences and developmental biology of the red flour beetle (Tribolium castaneum) across three food sources: broken rice with rice flour, broken wheat with wheat flour, and tapioca flour. Beetle development was observed in ventilated containers under controlled conditions (28-30°C, 65% relative humidity) with 25 beetles per container. Significant differences in beetle survival and development. Wheat flour was the most favourable host, with 304 live adults, followed by broken wheat (145 live adults. Rice flour supported 20 adults and 11 larvae, indicating moderate suitability. Whole rice grains and tapioca flour were the least effective, with only dead adults and few larvae. Despite host quality variations, the developmental stages (egg incubation: 4-7 days, larval: 65-70 days, pupal: 6-9 days) were consistent across food sources. Wheat flour is the most supportive environment for T. castaneum development.

Proteome-wide neuropeptide identification using NeuroPeptide-HMMer (NP-HMMer)

Neuropeptides are essential neuronal signaling molecules that orchestrate animal behavior and physiology via actions within the nervous system and on peripheral tissues. Due to the small size of biologically active mature peptides, their identification on a proteome-wide scale poses a significant challenge using existing bioinformatics tools like BLAST. To address this, we have developed NeuroPeptide-HMMer (NP-HMMer), a hidden Markov model (HMM)-based tool to facilitate neuropeptide discovery, especially in underexplored invertebrates. NP-HMMer utilizes manually curated HMMs for 46 neuropeptide families, enabling rapid and accurate identification of neuropeptides. Validation of NP-HMMer on Drosophila melanogaster, Daphnia pulex, Tribolium castaneum and Tenebrio molitor demonstrated its effectiveness in identifying known neuropeptides across diverse arthropods. Additionally, we showcase the utility of NP-HMMer by discovering novel neuropeptides in Priapulida and Rotifera, identifying 22 and 19 new peptides, respectively. This tool represents a significant advancement in neuropeptide research, offering a robust method for annotating neuropeptides across diverse proteomes and providing insights into the evolutionary conservation of neuropeptide signaling pathways.

Relationship between walking movement and reproductive traits in the red flour beetle Tribolium castaneum

AbstractMany animal species have movement abilities. Behavior is important for evolutionary ecology because animal movement leads to dispersal, migration, search for food and mates, and escaping from enemies. However, individual differences in movement activity are found within a population. This phenomenon can be affected by various factors, one of which is suggesting that higher moving activity has fitness cost, whereas lower moving activity has benefits. Animal movement may also affect reproduction (e.g., resource allocation tradeoff between movement and reproduction as well as intra‐ and intersexual selection). Although many previous studies have investigated the relationship between movement and reproduction, less attention has been paid to walking movement. In this study, previous studies that investigated the relationship between movement and reproduction were reviewed using the flour beetle Tribolium castaneum, a model insect in behavioral ecology and genetics. Several previous studies suggest that beetle walking is strongly associated with male and female reproductive traits. In recent years, empirical studies on the correlation between walking and other traits have increased, particularly in T. castaneum. Although this species can fly and walk, the movement is often discussed without discriminating between flight and walk. Differences in modes of movement may affect the correlation between movement and other traits; thus, discussing each mode of movement separately is necessary.

Tissue-Level Integration Overrides Gradations of Differentiating Cell Identity in Beetle Extraembryonic Tissue.

During animal embryogenesis, one of the earliest specification events distinguishes extraembryonic (EE) from embryonic tissue fates: the serosa in the case of the insects. While it is well established that the homeodomain transcription factor Zen1 is the critical determinant of the serosa, the subsequent realization of this tissue's identity has not been investigated. Here, we examine serosal differentiation in the beetle Tribolium castaneum based on the quantification of morphological and morphogenetic features, comparing embryos from a Tc-zen1 RNAi dilution series, where complete knockdown results in amnion-only EE tissue identity. We assess features including cell density, tissue boundary morphology, and nuclear size as dynamic readouts for progressive tissue maturation. While some features exhibit an all-or-nothing outcome, other key features show dose-dependent phenotypic responses with trait-specific thresholds. Collectively, these findings provide nuance beyond the known status of Tc-Zen1 as a selector gene for serosal tissue patterning. Overall, our approach illustrates how the analysis of tissue maturation dynamics from live imaging extends but also challenges interpretations based on gene expression data, refining our understanding of tissue identity and when it is achieved.

Effects of modified atmospheres, temperature and humidity on the growth and development of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) in feihe infant milk powder

Tribolium castaneum (Herbst) is a common stored product pest that damages various stored goods across the global. To identify the effects of modified atmospheres (MA), humidity and temperature on the growth and development of T. castaneum in Feihe infant milk powder, the modified atmospheres were set at 75% nitrogen (N2) and 25% carbon dioxide (CO2), with air as the control. Two different temperatures and humidities were set at 30 °C, RH 80% and 25 °C, RH 40%. The results revealed that T. castaneum larvae exposed to the warm and moist southern conditions (30 °C, RH 80%) feeding on flour and yeast diet had a significantly shorter developmental duration. The larval weights were significantly increased in T. castaneum feeding flour and yeast diet compared with those of T. castaneum feeding milk powder under high temperature. The elevated temperature significantly increased the mortality of larvae. However, 4.4% of the survival larvae successfully grow into adults at the cool and dry northern conditions (25 °C, RH 40%). The mortality rate of eggs was 100% when eggs were sealed and treated by MA under warm and moist temperature and humidity for 96 h, and the mortality rate of 5th instar larvae increased to 100% on 7th day. Moreover, the activities of CAT and GST were decreased in all stages of T. castaneum feeding on infant milk under temperature and humidity stress. Additionally, 5th instar larvae fed with infant milk powder showed significantly fluctuated activity of POD, SOD and P450 in all stages of T. castaneum. However, T. castaneum is more susceptible to milk powder sealing with air under warm and moist temperature and humidity. The present investigation demonstrated that temperature treatment and modified atmospheres effectively prevented growth and development of T. castaneum in milk powder. These results also contribute to suitable strategies for storage of milk power.

Exploring chemical composition and insecticidal activities of Alpinia calcarata rhizome essential oil against three major storage insects

Alpinia calcarata (Roscoe), generally recognized as snap ginger or lesser galangal of the Zingiberaceae family, undergoes widespread cultivation in tropical nations. It is characterized by its noteworthy pharmacological attributes, such as anti-inflammatory, analgesic, antinociceptive, reproductive, anti-cancer, and gastro-protective effects. Many storage pests now resist traditional chemical pesticides. Transitioning to organic alternatives is a preferable and sustainable solution. This research investigates the insecticidal activity of essential oil from A. calcarata rhizomes, obtained via hydro-distillation. Chemical analysis was conducted, and the study assessed its toxicity and repellent effects on Tribolium castaneum (Herbst), Lasioderma serricorne (F.), and Callosobruchus chinensis (L). A yield of 0.44 ± 0.02 % essential oil was obtained from dry rhizomes. Gas chromatography-mass spectrometry (GC-MS) characterization revealed the major chemical constituents as 1,8-cineole (25.71 %), ethyl iso-allocholate (16.01 %), and fenchyl acetate (15.15 %). The findings from the investigation into the pesticidal characteristics of essential oils derived from the rhizome of A. calcarata unveiled significant contact (LC50 at 24 h was 10.9 mg/cm2 for T. castaneum, 1.0 mg/cm2 for L. serricorne and 0.5 mg/cm2 for C. chinensis) and fumigant toxicity (LC50 at 24 h was 4.05 mg/L air for T. castaneum, 1.75 mg/L air for L. serricorne, and 1.03 mg/L air for C. chinensis). The repellent efficacy was evaluated through an area preference test, revealing robust repellent activity at various concentrations. Undoubtedly, the essential oil extracted from the rhizome of A. calcarata is valuable and has the potential to be a safe, organic pesticide for controlling storage pests.

Toxicity and morphological changes induced by diatomaceous earth & malathion against Tribolium castaneum

Toxicity and morphological changes induced by diatomaceous earth & malathion against Tribolium castaneum

Odourless vegetable oils as insect pest repellents for short-term protection of various food packaging materials

The repellent activity of two odourless vegetable oils (sweet almond—SAO and soybean—SO) applied on filter paper and four types of food packaging materials (polyethylene film, plastic gunnysacks, cardboard, and wood) was tested against three species of stored product pests (Sitophilus granarius, Tribolium castaneum, and Oryzaephilus surinamensis). The repellence of three doses was evaluated after 24 h, and after 7, 14, 21, and 28 days. Both oils exhibited a repellent effect when applied to filter paper, but this effect decreased with time and concentration. The most susceptible species was T. castaneum. The maximum repellence was 84% and 76% for SO and SAO, respectively. On packaging materials, the repellent effect diminished after a certain period of time, ranging from 7 to 28 days. Subsequently, the oils even became attractive. Generally, the repellent effects of both SO and SAO were higher on cardboard and wood slab than on polyethylene film and plastic gunnysacks. As such, in the case of SO, the repellence against T. castaneum was 88% 24 h after application on cardboard, whilst 68% on plastic bag. This work demonstrated the repellent potential of vegetable oils in the short term for the protection of packaged food against stored product pests.

Population growth of different stored product species on wheat, barley, and tritordeum

Stored product species can be developed in a variety of stored product grains. In this study, were compare three different grains for their suitability for population growth of primary and secondary stored product insects. In the first series, we evaluated the population growth and damage of Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae), Sitophilus granarius (L.) (Coleoptera: Curculionidae) and Sitophilus oryzae (L.) (Coleoptera: Curculionidae) on whole wheat, barley and tritordeum kernels after 65 days. In the second series, three different percentages of whole and cracked kernels were tested: 100% cracked kernels, 100% whole kernels and 50% cracked +50% whole kernels, for Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae) and Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). The results showed that tritordeum was suitable for the population growth of all species tested. In most of the cases, R. dominica showed higher frass production but S. granarius showed a higher number of damaged kernels. Cracked kernels were preferred by secondary pests than whole kernels and wheat showed the highest progeny production than tritordeum and barley. Overall, commodity, and grain type and percentage of cracked kernels can affect the population growth of stored product insect species. Results of this study showed that stored product insect species can develop in tritordeum and thus should be taken into consideration for further results as may be damaged during storage.

Chemical Compositions and Fumigation Effects of Essential Oils Derived from Cardamom, Elettaria cardamomum (L.) Maton, and Galangal, Alpinia galanga (L.) Willd, against Red Flour Beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae).

This study explores the use of essential oils from cardamom (Elettaria cardamomum (L.) Maton) and galangal (Alpinia galanga (L.) Willd) as alternatives to synthetic insecticides for controlling the red flour beetle, Tribolium castaneum (Herbst). The chemical compositions of these oils were analyzed using GC-MS, and their fumigation effects were tested in a vapor-phase bioassay. The experiment followed a factorial design with four types of essential oils, namely, those manually extracted from cardamom leaves (MCL) and galangal leaves (MGL) and those commercially produced from cardamom seeds (CCS) and galangal rhizomes (CGR), at seven concentrations (0, 50, 100, 150, 200, 250, and 300 µL/L air). The manually extracted oils yielded 0.6% from cardamom leaves and 0.25% from galangal leaves. MCL contained 28 components, with eucalyptol (25.2%) being the most abundant, while CCS had 34 components, primarily α-terpinyl acetate (46.1%) and eucalyptol (31.2%). MGL included 25 components, mainly caryophyllene (28.7%) and aciphyllene (18.3%), whereas CGR comprised 27 components, with methyl cis-cinnamate (47.3%) and safrole (19.8%) as the major constituents. The fumigation bioassay results revealed that CGR was the most effective, demonstrating the highest mortality rates of T. castaneum across all the tested periods and concentrations, achieving up to 96% mortality at 168 h with a concentration of 300 µL/L air. Statistical analyses showed significant differences in mortality based on the type and concentration of essential oil, particularly after 96 h. These findings highlight the potential of CGR, with its advantages and differences in chemical composition, as an effective biopesticide against T. castaneum, with increasing efficacy over time and at higher concentrations.

Behavioral events and functional analysis of bursicon signal during adult eclosion in the 28-spotted larger potato ladybird

To accommodate growth, insects must periodically shed their exoskeletons. In Manduca sexta, Drosophila melanogaster and Tribolium castaneum, Bursicon (Burs)/ Partner of bursicon (Pburs)-LGR2 signal is an indispensable component for the proper execution of ecdysis behavior during adult eclosion. Nevertheless, the behavioral events and the roles of bursicon signaling in other insects deserve further exploration. In the current paper, we found that the pupal-adult ecdysis in Henosepilachna vigintioctomaculata could be divided into three distinct stages, preecdysis, ecdysis and postecdysis. Preecdysis behavioral sequences included abdomen twitches, dorsal-ventral contractions and air filling that function to loosen the old cuticle. Ecdysis events began with anterior-posterior contractions that gradually split the old integument along the dorsal body midline, followed by freeing of legs and mouthparts, and culminated in detachment from pupal cuticle. Postecdysis behavioral processes contained three actions: perch selection and stretching of elytra and hindwings. RNA interference for HvBurs, HvPburs or Hvrk (encoding LGR2) strongly impaired wing expansion actions, and slightly influenced preecdysis and ecdysis behaviors. The RNAi beetles failed to extend their elytra and hindwings. In addition, injected with dsrk also caused kinked femurs and tibia. Our findings establish that bursicon pathway is involved in regulation of adult eclosion behavior, especially wing expansion motor programs. Given that wings facilitate food foraging, courtship, predator avoidance, dispersal and migration, our results provide a potential target for controlling H. vigintioctomaculata.

Efficiency of Alkaloid and Phenolic Extracts of Seeds and Leaves Sesbania Plant on Adults Red Flour Beetle Tribolium castaneum Herbst (Coleoptera : Tenebrionidae)

Efficiency of Alkaloid and Phenolic Extracts of Seeds and Leaves Sesbania Plant on Adults Red Flour Beetle Tribolium castaneum Herbst (Coleoptera : Tenebrionidae)

Evaluating Effectiveness of Some Inert Dusts and Chemical Pesticides in Tribolium castaneum Coleoptera: Tenebrindae Under Laboratory Conditions

Evaluating Effectiveness of Some Inert Dusts and Chemical Pesticides in Tribolium castaneum Coleoptera: Tenebrindae Under Laboratory Conditions

Characterization of double-stranded RNA and its silencing efficiency for insects using hybrid deep-learning framework.

RNA interference (RNAi) technology is widely used in the biological prevention and control of terrestrial insects. One of the main factors with the application of RNAi in insects is the difference in RNAi efficiency, which may vary not only in different insects, but also in different genes of the same insect, and even in different double-stranded RNAs (dsRNAs) of the same gene. This work focuses on the last question and establishes a bioinformatics software that can help researchers screen for the most efficient dsRNA targeting target genes. Among insects, the red flour beetle (Tribolium castaneum) is known to be one of the most sensitive to RNAi. From iBeetle-Base, we extracted 12027 efficient dsRNA sequences with a lethality rate of ≥20% or with experimentation-induced phenotypic changes and processed these data to correspond to specific silence efficiency. Based on the first complied novel benchmark dataset, we specifically designed a deep neural network to identify and characterize efficient dsRNA for RNAi in insects. The dna2vec word embedding model was trained to extract distributed feature representations, and three powerful modules, namely convolutional neural network, bidirectional long short-term memory network, and self-attention mechanism, were integrated to form our predictor model to characterize the extracted dsRNAs and their silencing efficiencies for T. castaneum. Our model dsRNAPredictor showed reliable performance in multiple independent tests based on different species, including both T. castaneum and Aedes aegypti. This indicates that dsRNAPredictor can facilitate prescreening for designing high-efficiency dsRNA targeting target genes of insects in advance.