Mapping the nutritional landscape in the yellow mealworm: testing the nutrient-mediated life-history trade-offs.

Animals must acquire an ideal amount and balance of macronutrients to optimize their performance, health and fitness. The nutritional landscape provides an integrative framework for analysing how animal phenotypes are associated with multiple nutritional components. Here, we applied this powerful approach to examine how the intake of protein and carbohydrate affects nutrient acquisition and performance in the yellow mealworm (Tenebrio molitor) reared on one of 42 synthetic foods varying in protein and carbohydrate content. Tenebrio molitor larvae increased their food consumption rate in response to nutrient dilution, but this increase was not sufficient to fully compensate for the dilution. Diluting the food nutrient content with cellulose reduced the efficiency of post-ingestive nutrient utilization, further restricting macronutrient acquisition. Tenebrio molitor larvae utilized macronutrients most efficiently at a protein to carbohydrate (P:C) ratio of 1.77:1, but became less efficient at imbalanced P:C ratios. Survivorship was high at high protein intake and fell with decreasing protein intake. Pupal mass and growth rate exhibited a bell-shaped landscape, with the nutritional optima being located around protein-biased P:C ratios of 1.99:1 to 2.03:1 and 1.66:1 to 2.86:1, respectively. The nutritional optimum for development time was also identified at high P:C ratios (1.66:1  to 5.86:1). Unlike these performance traits, lipid content was maximized at carbohydrate-biased P:C ratios of 1:3.88 to 1:3.06. When given a food choice, T. molitor larvae self-composed a slightly carbohydrate-biased P:C ratio of 1:1.24, which lies between the P:C ratios that maximize performance and lipid content. Our findings indicate the occurrence of a nutrient-mediated trade-off between performance and energy storage in this insect.

Measurement of oxygen consumption in Tenebrio molitor using a sensitive, inexpensive, sensor-based coulometric microrespirometer.

Coulometric respirometry is a highly sensitive method for measuring O2 consumption in small organisms but it is not in widespread use among physiologists. Here, we describe a coulometric microrespirometer based on a digital environmental sensor inside a sealed glass chamber and controlled by an Arduino™ microcontroller. As O2 is consumed, exhaled CO2 is removed, causing pressure to decrease in the chamber. The sensor detects the decreased pressure, and the controller activates electrolytic production of O2, returning pressure to the initial value. O2 consumption is calculated from electrolytic charge transfer. The effects of developmental stage, body mass and temperature on O2 consumption of Tenebrio molitor beetles were easily measured by the apparatus. This straightforward design is a significant innovation in that it provides continuous data regarding environmental conditions inside the experimental chamber, can be fabricated easily, and is adaptable to a wide range of uses.

Gastric canthariasis caused by invasion of mealworm beetle larvae in weaned pigs in large-scale farming.

BACKGROUND: Mealworm beetle T. molitor (Coleoptera: Tenebrionidae) (Linnaeus, 1758) is one of the most important cosmopolitan primary storage pests, scavenging on a variety of post-harvest grains and affecting the quality and safety of food and feed. In addition to being an important factor in feed hygiene, the insect can also be an epidemiological factor of canthariasis. Livestock infestations with T. molitor are rarely reported. This article describes T. molitor-caused canthariasis in pigs in large scale closed-cycle farming. RESULTS: In the spring, we registered a significantly increased mortality among weaned pigs. In autopsy, live 3-6 mm long T. molitor larvae were found in their stomachs, especially in the non-glandular oesophageal region, on average 2-3 larvae per 10 cm2 of gastric mucosa. Corrective actions reduced the number of deaths back to basal levels. CONCLUSIONS: This is the first documented case of potentially lethal gastric canthariasis in weaned pigs, caused by invasion of T. molitor larvae. Although canthariasis caused by T. molitor has not been a significant problem in farm animals so far, our case indicates that the presence of mealworm beetles is a potential threat to animal welfare and health.

Development and Biomass Composition of Ephestia kuehniella (Lepidoptera: Pyralidae), Tenebrio molitor (Coleoptera: Tenebrionidae), and Hermetia illucens (Diptera: Stratiomyidae) Reared on Different Byproducts of the Agri-Food Industry.

The aim of this study was to evaluate five agro-industrial byproducts (apricots, brewer's spent grains, brewer's spent yeast, feed mill byproducts including broken cereal grains, and hatchery waste including eggshell debris, fluff, infertile eggs, dead embryos, and egg fluids) or mixtures thereof as food diets of Ephestia kuehniella (Zeller), Tenebrio molitor (L.), and Hermetia illucens (L.). Eleven out of 26 tested combinations allowed the first instar larvae to reach the adult stage. Results showed that bioconversion parameters and biomass composition can vary depending on the diet composition, especially in the case of E. kuehniella and H. illucens, whose nutritional requirements seem more complex than those of T. molitor. Tenebrio molitor was able to develop in almost all byproducts. However, only when T. molitor was fed with suitable mixtures of byproducts the development parameters were similar to those obtained with the standard diet. The best results in terms of bioconversion parameters were obtained by feeding H. illucens with a diet including dried brewer's spent grain, feed mill byproducts and brewer's spent yeast. The larvae of these three species can be considered interesting from a nutritional point of view, because of their high protein and fat content. However, the fatty acids profile of H. illucens larvae, with high proportions of saturated fatty acids, seems less healthy for human consumption compared with those of E. kuehniella and T. molitor.

Nutritional, Physiochemical, and Antioxidative Characteristics of Shortcake Biscuits Enriched with Tenebrio molitor Flour.

Edible insects, due to their high nutritional value, are a good choice for traditional food supplementation. The effects of partial replacement of wheat flour and butter with mealworm flour (Tenebrio molitor) on the quality attributes of shortcake biscuits were studied. The approximate composition was analyzed, along with the physical properties and color. Moreover, the antioxidant properties, starch digestibility, and glycemic index were determined in vitro. The protein and ash contents in biscuits supplemented with mealworm flour increased, while the carbohydrates content decreased. The increasing insect flour substitution decreased the lightness (L*) and yellowness (b*) but increased the redness (a*), total color difference (ΔE), and browning index (BI). The spread factor for the sample with the highest proportion of mealworm flour was significantly higher than the other biscuits. Furthermore, higher additions of mealworm flour increased the antioxidant activity of the biscuits and contributed to an increase in the content of slowly digested starch, with a decrease in the content of rapidly digested starch. Therefore, the results of the research are promising and indicate the possibility of using edible insects to enrich food by increasing the nutritional and health-promoting values.

Body Mass Increase Induced by Eight Years of Artificial Selection in the Yellow Mealworm (Coleoptera: Tenebrionidae) and Life History Trade-offs.

Efforts to improve rearing conditions of Tenebrio molitor L. (Coleoptera: Tenebrionidae) for insect biomass production included selecting for larger size pupae. The effects of an 8-yr continuous selection of T. molitor pupae for larger size were studied. Data consisting of daily counts and weights of pupae were analyzed using regression to determine the effects of selection over time. A preliminary evaluation of food conversion, growth, fecundity, and larval survival was done to compare ancestral versus selected strains. A significant positive correlation was identified between pupal size and time indicating a significant increase in pupal size over time in the selected T. molitor strain. A preliminary comparison of ancestral and selected strains showed significantly larger pupal size, growth rate, fecundity, and efficiency of conversion of ingested food in the selected strain. However, the selected strain also showed significantly lower larval survival than the ancestral strain. The low larval survival impacted the overall productivity of the selected strain resulting in no significant differences in biomass production when compared with the ancestral strain. The potential of using selection to improve biomass productivity in T. molitor is discussed.

Heart mechanical and hemodynamic parameters of a beetle, Tenebrio molitor, at selected ages.

The physiological processes that occur during the aging of insects are poorly understood. The aim of this study was to describe the changes in contractile activity and hemodynamic parameters of the heart that take place as the coleopteran beetle, Tenebrio molitor, ages. The frequency of heart contractions in beetles that had just undergone metamorphosis (median 24.7 beats/min) was significantly lower than the frequency of heart contractions in older beetles. In 56% of beetles that were < 1 week of age, a pattern of contractile activity with alternating periods of higher and lower contraction frequency was detected, suggesting that some posteclosion developmental processes occur during the first week of adulthood. All beetles that were 1 week of age showed a regular rhythm of heart contractions (median 72 beats/min). In older beetles, abnormalities such as heart arrhythmias or heart arrest were observed. The incidence of arrhythmia as well as the arrhythmicity index was highest in beetles that were 8-18 weeks old. The calculated stroke volume (SV) was also found to increase from eclosion to 12 weeks of age, and then decreased as adults aged further. Interestingly, cardiac output increased gradually, but the ejection fraction did not change significantly with age.

Microbial Hydrolysis of Racemic ß-Aryl-γ-ethylidene-γ-lactones and Antifeedant Activity of the Products against Alphitobius diaperinus Panzer.

Hydrolysis of (±)-ß-aryl-γ-ethylidene-γ-lactones by fungal strain Aspergillus ochraceus AM370 afforded (−)-(S)-γ-ethylidene-γ-lactones 2a⁻d and (+)-(R)-γ-ketoacids 3a⁻d. Enantiomeric purity of the unreacted lactones was strictly related to a size of an aryl substituent at C-4 of γ-lactone ring, with the highest ee (77%) obtained for the (−)-(S)-γ-ethylidene-γ-lactone possessing unsubstituted benzene ring (2a) and the lowest one (15%) determined for the (−)-(S)-γ-ethylidene-γ-lactone with bulky 1,3-benzodioxole system (2d). Lactones 2a⁻d, both racemic and enantiomerically enriched, as well as products of their hydrolysis showed varying degrees of feeding deterrent activity against lesser mealworm, Alphitobius diaperinus Panzer, which depended on the structure of the compound and the developmental stage of the lesser mealworm. In the case of adults, more active were γ-lactones 2a⁻d, compared with ketoacids 3a⁻d. Only in the case of lactone 2a was the effect of configuration of stereogenic center on the activity found. Particularly strong deterrents against this stage (T > 180) were racemic and (−)-(S)-γ-ethylidene-γ-lactone with p-methoxysubstituted phenyl ring (2c).

Temperature-driven plasticity in nutrient use and preference in an ectotherm.

Environmental temperature has strong effects on the rate and efficiency of resource use in ectotherms, but little is known about how changes in temperature influence their diet selection patterns. Changes in temperature may alter the balance of nutrients required by ectotherms by affecting metabolism. In response to temperature changes, ectotherms are predicted to express a preference for a specific nutrient (protein or carbohydrate) to match their altered nutrient requirement. Here, we examined the nutritional consequences of mealworm beetles (Tenebrio molitor L.) that were constrained to diets varying in protein:carbohydrate balance (P:C = 1:5, 1:1, or 5:1) or offered a choice between two nutritionally complementary diets (1:5 vs. 5:1) at four different temperatures (20, 25, 30, or 35 °C). Beetles had high mortality and reduced body mass at higher temperatures. Post-ingestive use efficiencies of both protein and carbohydrate decreased as temperature rose. Warming-driven decrease in carbohydrate use efficiencies occurred most profoundly when carbohydrates were consumed excessively. When given a choice, beetles selected protein and carbohydrate equally at 25 and 30 °C, but exhibited a significant preference for carbohydrate at 35 °C. Since carbohydrate is an immediate source of energy, this warming-driven preference for carbohydrate is explicable as an adaptive response of beetles to meet increased energy needs at high temperature. Beetles exposed to 20 °C ate substantially less food, but preferentially consumed carbohydrate over protein possibly to cope with reduced energy intake. The present findings have implications for the impact of temperature on foraging and resource use in ectotherms.

Characterization of T-DNA insertion mutants with decreased virulence in the entomopathogenic fungus Beauveria bassiana JEF-007.

The bean bug, Riptortus pedestris, is a major agricultural pest that reduces crop quality and value. Chemical pesticides have contributed to pest management, but resistance to these chemicals has significantly limited their use. Alternative strategies with different modes of action, such as entomopathogenic fungi, are therefore of great interest. Herein, we explored how entomopathogenic fungi can potentially be used to control the bean bug and focused on identifying virulence-related genes. Beauveria bassiana (JEF isolates) were assayed against bean bugs under laboratory conditions. One isolate, JEF-007, showed >80 % virulence by both spray and contact exposure methods. Agrobacterium tumefaciens-mediated transformation (AtMT) of JEF-007 generated 249 random transformants, two of which (B1-06 and C1-49) showed significantly reduced virulence against Tenebrio molitor and R. pedestris immatures. Both species were used for rapid screening of virulence-reduced mutants. The two transformants had different morphologies, conidial production, and thermotolerance than the wild type. To determine the localization of the randomly inserted T-DNA, thermal asymmetric interlaced (TAIL) PCR was conducted and analysis of the two clones found multiple T-DNA insertions (two in B1-06 and three in C1-49). Genes encoding complex I intermediate-associated protein 30 (CIA30) and the autophagy protein (Atg22) were possibly disrupted by the T-DNA insertion and might be involved in the virulence. This work provides a strong platform for future functional genetic studies of bean bug-pathogenic B. bassiana. The genes putatively involved in fungal virulence should be experimentally validated by knockdown in future studies.

Functional analysis of C1 family cysteine peptidases in the larval gut of Тenebrio molitor and Tribolium castaneum.

BACKGROUND: Larvae of the tenebrionids Tenebrio molitor and Tribolium castaneum have highly compartmentalized guts, with primarily cysteine peptidases in the acidic anterior midgut that contribute to the early stages of protein digestion. RESULTS: High throughput sequencing was used to quantify and characterize transcripts encoding cysteine peptidases from the C1 papain family in the gut of tenebrionid larvae. For T. castaneum, 25 genes and one questionable pseudogene encoding cysteine peptidases were identified, including 11 cathepsin L or L-like, 11 cathepsin B or B-like, and one each F, K, and O. The majority of transcript expression was from two cathepsin L genes on chromosome 10 (LOC659441 and LOC659502). For cathepsin B, the major expression was from genes on chromosome 3 (LOC663145 and LOC663117). Some transcripts were expressed at lower levels or not at all in the larval gut, including cathepsins F, K, and O. For T. molitor, there were 29 predicted cysteine peptidase genes, including 14 cathepsin L or L-like, 13 cathepsin B or B-like, and one each cathepsin O and F. One cathepsin L and one cathepsin B were also highly expressed, orthologous to those in T. castaneum. Peptidases lacking conservation in active site residues were identified in both insects, and sequence analysis of orthologs indicated that changes in these residues occurred prior to evolutionary divergence. Sequences from both insects have a high degree of variability in the substrate binding regions, consistent with the ability of these enzymes to degrade a variety of cereal seed storage proteins and inhibitors. Predicted cathepsin B peptidases from both insects included some with a shortened occluding loop without active site residues in the middle, apparently lacking exopeptidase activity and unique to tenebrionid insects. Docking of specific substrates with models of T. molitor cysteine peptidases indicated that some insect cathepsins B and L bind substrates with affinities similar to human cathepsin L, while others do not and have presumably different substrate specificity. CONCLUSIONS: These studies have refined our model of protein digestion in the larval gut of tenebrionid insects, and suggest genes that may be targeted by inhibitors or RNA interference for the control of cereal pests in storage areas.

Biodegradation and Mineralization of Polystyrene by Plastic-Eating Mealworms: Part 1. Chemical and Physical Characterization and Isotopic Tests.

Polystyrene (PS) is generally considered to be durable and resistant to biodegradation. Mealworms (the larvae of Tenebrio molitor Linnaeus) from different sources chew and eat Styrofoam, a common PS product. The Styrofoam was efficiently degraded in the larval gut within a retention time of less than 24 h. Fed with Styrofoam as the sole diet, the larvae lived as well as those fed with a normal diet (bran) over a period of 1 month. The analysis of fecula egested from Styrofoam-feeding larvae, using gel permeation chromatography (GPC), solid-state (13)C cross-polarization/magic angle spinning nuclear magnetic resonance (CP/MAS NMR) spectroscopy, and thermogravimetric Fourier transform infrared (TG-FTIR) spectroscopy, substantiated that cleavage/depolymerization of long-chain PS molecules and the formation of depolymerized metabolites occurred in the larval gut. Within a 16 day test period, 47.7% of the ingested Styrofoam carbon was converted into CO2 and the residue (ca. 49.2%) was egested as fecula with a limited fraction incorporated into biomass (ca. 0.5%). Tests with α (13)C- or ß (13)C-labeled PS confirmed that the (13)C-labeled PS was mineralized to (13)CO2 and incorporated into lipids. The discovery of the rapid biodegradation of PS in the larval gut reveals a new fate for plastic waste in the environment.

Biodegradation and Mineralization of Polystyrene by Plastic-Eating Mealworms: Part 2. Role of Gut Microorganisms.

The role of gut bacteria of mealworms (the larvae of Tenebrio molitor Linnaeus) in polystyrene (PS) degradation was investigated. Gentamicin was the most effective inhibitor of gut bacteria among six antibiotics tested. Gut bacterial activities were essentially suppressed by feeding gentamicin food (30 mg/g) for 10 days. Gentamicin-feeding mealworms lost the ability to depolymerize PS and mineralize PS into CO2, as determined by characterizing worm fecula and feeding with (13)C-labeled PS. A PS-degrading bacterial strain was isolated from the guts of the mealworms, Exiguobacterium sp. strain YT2, which could form biofilm on PS film over a 28 day incubation period and made obvious pits and cavities (0.2-0.3 mm in width) on PS film surfaces associated with decreases in hydrophobicity and the formation of C-O polar groups. A suspension culture of strain YT2 (10(8) cells/mL) was able to degrade 7.4 ± 0.4% of the PS pieces (2500 mg/L) over a 60 day incubation period. The molecular weight of the residual PS pieces was lower, and the release of water-soluble daughter products was detected. The results indicated the essential role of gut bacteria in PS biodegradation and mineralization, confirmed the presence of PS-degrading gut bacteria, and demonstrated the biodegradation of PS by mealworms.

Resource availability as a proxy for terminal investment in a beetle.

Terminal investment hypothesis is a longstanding theoretical idea that organisms should increase their reproductive effort as their prospects for survival and reproduction decline. However, numerous attempts to test the terminal investment in reproduction have yielded contradictory results. This study reports an experimental confirmation of the terminal investment hypothesis. It was predicted that immune-challenged yellow mealworm beetles (Tenebrio molitor) are more likely to follow terminal investment strategy when their food resources are limited. Our results suggest the key role of food resources while making decisions to follow a terminal investment strategy. We found that male individuals invested in their sexual attractiveness at the expense of immune response and survival when food was not available. In contrast, the beetles did not decrease their lifespan and did not invest in the attractiveness of their sex odours under conditions of food ad libitum. Our results show the importance of food availability and quality in understanding the evolution of reproductive strategies.

Effect of Larval Density on Food Utilization Efficiency of Tenebrio molitor (Coleoptera: Tenebrionidae).

Crowding conditions of larvae may have a significant impact on commercial production efficiency of some insects, such as Tenebrio molitor L. (Coleoptera: Tenebrionidae). Although larval densities are known to affect developmental time and growth in T. molitor, no reports were found on the effects of crowding on food utilization. The effect of larval density on food utilization efficiency of T. molitor larvae was studied by measuring efficiency of ingested food conversion (ECI), efficiency of digested food conversion (EDC), and mg of larval weight gain per gram of food consumed (LWGpFC) at increasing larval densities (12, 24, 36, 48, 50, 62, 74, and 96 larvae per dm(2)) over four consecutive 3-wk periods. Individual larval weight gain and food consumption were negatively impacted by larval density. Similarly, ECI, ECD, and LWGpFC were negatively impacted by larval density. Larval ageing, measured as four consecutive 3-wk periods, significantly and independently impacted ECI, ECD, and LWGpFC in a negative way. General linear model analysis showed that age had a higher impact than density on food utilization parameters of T. molitor larvae. Larval growth was determined to be responsible for the age effects, as measurements of larval mass density (in grams of larvae per dm(2)) had a significant impact on food utilization parameters across ages and density treatments (in number of larvae per dm(2)). The importance of mass versus numbers per unit of area as measurements of larval density and the implications of negative effects of density on food utilization for insect biomass production are discussed.

The natural insect peptide Neb-colloostatin induces ovarian atresia and apoptosis in the mealworm Tenebrio molitor.

BACKGROUND: The injection of Neb-colloostatin into T. molitor females causes gonadoinhibitory effects on ovarian development. This peptide inhibits intercellular space formation (patency) in follicular epithelium and results in slowed vitellogenesis, delayed ovulation, reduced number of eggs laid and presumably cell death in the terminal follicles. However, as does the form of cell death in the terminal follicle, the mode of action of Neb-colloostatin remains unknown. RESULTS: We tested Neb-colloostatin for a sterilizing effect on females of Tenebrio molitor. We report that injection of nanomolar doses of Neb-colloostatin induce ovarian follicle atresia in 4-day old females during their first gonadotropic cycle. Light microscope observations revealed morphological changes in the ovary: after Neb-colloostatin injection the terminal oocytes are significantly smaller and elicit massive follicle resorption, but the control terminal follicles possess translucent ooplasm in oocytes at different stages of vitellogenesis. A patency is visible in follicular epithelium of the control vitellogenic oocytes, whereas peptide injection inhibits intercellular space formation and, in consequence, inhibits vitellogenesis. Confocal and electron microscope examination showed that peptide injection causes changes in the morphology indicating death of follicular cells. We observed F-actin cytoskeleton disorganization, induction of caspase activity, changes in chromatin organization and autophagic vacuole formation. Moreover, the apical cytoplasm of follicular cells is filled with numerous free ribosomes, probably indicating a higher demand for protein biosynthesis, especially in preparation for autophagic vacuole formation. On the other hand, the process of polyribosomes formation is inhibited, indicating the contributing effect of this hormone. CONCLUSION: Neb-colloostatin induces atresia in the mealworm ovary. Degeneration of T. molitor follicles includes changes in morphology and viability of follicular cells, and oosorption as a consequence of these changes.

Efficacy of Metarhizium anisopliae isolate MAX-2 from Shangri-la, China under desiccation stress.

BACKGROUND: Metarhizium anisopliae, a soil-borne entomopathogen found worldwide, is an interesting fungus for biological control. However, its efficacy in the fields is significantly affected by environmental conditions, particularly moisture. To overcome the weakness of Metarhizium and determine its isolates with antistress capacity, the efficacies of four M. anisopliae isolates, which were collected from arid regions of Yunnan Province in China during the dry season, were determined at different moisture levels, and the efficacy of the isolate MAX-2 from Shangri-la under desiccation stress was evaluated at low moisture level. RESULTS: M. anisopliae isolates MAX-2, MAC-6, MAL-1, and MAQ-28 showed gradient descent efficacies against sterile Tenebrio molitor larvae, and gradient descent capacities against desiccation with the decrease in moisture levels. The efficacy of MAX-2 showed no significant differences at 35% moisture level than those of the other isolates. However, significant differences were found at 8% to 30% moisture levels. The efficacies of all isolates decreased with the decrease in moisture levels. MAX-2 was relatively less affected by desiccation stress. Its efficacy was almost unaffected by the decrease at moisture levels > 25%, but slowly decreased at moisture levels < 25%. By contrast, the efficacies of other isolates rapidly decreased with the decrease in moisture levels. MAX-2 caused different infection characteristics on T. molitor larvae under desiccation stress and in wet microhabitat. Local black patches were found on the cuticles of the insects, and the cadavers dried without fungal growth under desiccation stress. However, dark black internodes and fungal growth were found after death of the insects in the wet microhabitat. CONCLUSIONS: MAX-2 showed significantly higher efficacy and superior antistress capacity than the other isolates under desiccation stress. The infection of sterile T. molitor larvae at low moisture level constituted a valid laboratory bioassay system in evaluating M. anisopliae efficacy under desiccation stress.

Better the devil you know: avian predators find variation in prey toxicity aversive.

Toxic prey that signal their defences to predators using conspicuous warning signals are called 'aposematic'. Predators learn about the toxic content of aposematic prey and reduce their attacks on them. However, through regulating their toxin intake, predators will include aposematic prey in their diets when the benefits of gaining the nutrients they contain outweigh the costs of ingesting the prey's toxins. Predators face a problem when managing their toxin intake: prey sharing the same warning signal often vary in their toxicities. Given that predators should avoid uncertainty when managing their toxin intake, we tested whether European starlings (Sturnus vulgaris) preferred to eat fixed-defence prey (where all prey contained a 2% quinine solution) to mixed-defence prey (where half the prey contained a 4% quinine solution and the other half contained only water). Our results support the idea that predators should be more 'risk-averse' when foraging on variably defended prey and suggest that variation in toxicity levels could be a form of defence.

Adaptation of the egg of the desert beetle, Microdera punctipennis (Coleoptera: Tenebrionidae), to arid environment.

Microdera punctipennis Kaszab (Coleoptera: Tenebrionidae) is an endemic species in Guerbantonggut desert in China. To explore the ways that M. punctipennis egg adapts to dry desert environment, morphological characteristics of the egg was investigated along with the egg of the nondesert beetle Tenebrio molitor (Coleoptera: Tenebrionidae). Water loss rate and survival rate of these eggs under different dry treatments (relative humidity0, 10, and 20%) were measured to evaluate the desiccation resistance of the eggs at different developmental stages (day 0, 2, and 5 eggs). Our results showed that the 50-egg weight in T. molitor was heavier than M. punctipennis, while the 50-first-instar larva weight in T. molitor was almost the same as in M. punctipennis. The water loss rate of M. punctipennis egg under dry conditions was significantly lower than T. molitor, and the egg survival rate was significantly higher than T. molitor. The estimated developmental threshold temperature of M. punctipennis egg was 18.30°C, and the critical thermal maximum of M. punctipennis egg is above 39°C. These features partly account for the adaptability of M. punctipennis to desert environment in egg stage.

Goal-directed behavior in Tenebrio molitor larvae.

Can signs of intentional behavior be traced in an insect larva, traditionally thought to be driven only by mere reflexes? We trained Tenebrio molitor coleoptera larvae in a uniform Y-maze to prefer one target branch to get access to food, observing their ability to learn and retain access to the reward-associated side for up to 24 h. During reward devaluation, the reward food (experimental group) and a different food (control group) were paired with an aversive stimulus in a new environment. When tested again in the Y-maze, mealworms of the experimental group significantly reduced their visits to the target branch, whereas mealworms of the control group did not. Importantly, we found that the larvae did not have to experience the unpleasant consequences directly in the target branch to halt their behavior, as the exposure to the aversive taste occurred in a separate unfamiliar context. This is evidence that the mealworms formed a mental representation of action-consequence relationships, demonstrating flexible control of their actions to achieve desired outcomes at an early stage of their development.