Artificial Diet Research Articles

The metabolism and detoxification effects of lead exposure on Aleurolyphus ovatus (Acari: Acaridae) via transcriptome analysis

Aleurolyphus ovatus Troupeau is one of the most predominant species of the Acaridae family worldwide. Recent reports have demonstrated that the accumulation of lead in stored grains and dietary items exceeds the required standards. However, the molecular mechanism of heavy metal stress on mites has not been reported. To understand the mechanism underlying the heavy metal response of A. ovatus, comparative transcriptome analysis was performed in this study using an Illumina high throughput mRNA sequencing (RNA-seq) platform. A. ovatus was fed on artificial diets containing two different concentrations of lead, namely, a low concentration of 12.5 mg/kg (LAO) and a high concentration of 100 mg/kg (HAO), while the mites in the control (NAO) group were not exposed to lead. A total of 44,362 unigenes, with an average length of 1547 bp, were identified. Of these, 996 unigenes were successfully annotated in seven functional databases. The number of differentially expressed genes (DEGs) in A. ovatus under different lead concentrations was compared. In NAO versus LAO group, including 310 up-regulated and 1580 down-regulated DEGs. In NAO versus HAO group, including 3928 up-regulated and 1761 down-regulated DEGs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment indicated that detoxification enzyme genes were significantly expressed in pathways, such as cytochrome P450 foreign body metabolism, glutathione metabolism and drug metabolism-cytochrome pathway. The results of gene annotation and quantitative real-time PCR showed that high concentration of lead significantly stimulated the expression of metabolic detoxification enzyme genes such as glutathione S transferase (GST) and superoxide dismutase (SOD), while low concentration inhibited their expression. This study will provide a basis for the molecular mechanism of A. ovatus in response to heavy metal lead stimulation in stored grain.

Monitoring of insecticide resistance for Apolygus lucorum populations in the apple orchard in China

Apolygus lucorum (Meyer-Dur) (Hemiptera: Miridae) is an important piercing-sucking pest that has been recorded on more than 200 host plants in China. In recent years, its damage to apple trees has become more and more serious, resulting in malformed fruit and even fruit shedding. It is primarily controlled with insecticides. However, the current status of insecticide resistance of A. lucorum in the apple orchard is rarely reported. In this study, we determined the susceptibility of one laboratory population and six geographic field populations of A. lucorum to ten chemical and six botanical insecticides by glass-vial and artificial diet bioassays. The results showed that among the ten chemical insecticides, bifenthrin exhibited the highest toxicity to this pest by glass-vial bioassays than other insecticides, followed by chlorpyrifos, and then one of the neonicotinoid insecticides nitenpyram. In the artificial diet bioassays, the LC50 values of the avermectin insecticides to A. lucorum populations were significantly lower than those of other classes of insecticides, followed by neonicotinoid insecticides. Among six botanical pesticides, veratramine and pyrethrins showed higher toxicity to A. lucorum populations, while rotenone and cnidiadin exhibited poor effects. The results indicated that chlorpyrifos, bifenthrin, avermectin, and emamectin benzoate could be recommended to mix or rotate with botanical pesticides veratramine and pyrethrins in controlling A. lucorum in the apple orchard. Our study would provide valuable information for choosing alternative insecticides and for insecticide resistance management of A. lucorum.

Transcriptome analysis of anorexic and preferred silkworms (Bombyx mori) on artificial diet

The silkworm, Bombyx mori, is an important oligophagous economic insect and feeding habits of different silkworm varieties to artificial diet are different. Research on the mechanisms of feeding habits on artificial diet, and breeding of silkworm varieties adapted on artificial diet, which is a necessary condition for industrial silkworm rearing, is currently lacking. For an artificial diet, Xin was anorexic, whereas Haoyue A showed a strong appetite. When the two varieties were crossed, the F1 generation showed a poor appetite for the artificial diet and had a setae dispersion rate of <50 %. However, the F2 generation, self-bred progeny of F1, had a good appetite for the artificial diet, with a setae dispersion rate of 70 %. Herein, transcriptome analysis was conducted on the F2 generation, comparing individuals with anorexic and preferred feeding habits, and 2188 differential genes were identified, with 1524 genes up-regulated and 934 genes down-regulated. Several genes were identified to contribute to feeding habits, such as genes involved olfactory system, energy supply, and cell proliferation and differentiation. GO enrichment revealed a large number of DEGs related to behavior, growth, signaling, developmental process, response to stimulation, and other pathways. Furthermore, proteins closely related to feeding were expressed differently. Some DEGs were selected for qRT-PCR, and results indicated the reliability of the DEG results. The DEGs between individuals with anorexic and preferred feeding habits were screened by RNA-Seq technology, which provides a reliable reference to study molecule mechanisms of feeding habits on artificial diet.

Optimizing Mass Rearing of the Egg Parasitoid, Telenomus podisi, for Control of the Brown Stink Bug, Euschistus heros

The parasitoid Telenomus podisi Ashmead, 1893 is used in biological control programs in Brazil against eggs of Euschistus heros (Fabricius, 1798), a key pest of soybean, Glycine max (L.) Merr. To optimize the mass production of parasitoids, artificial diets and storage of host eggs at low temperatures have been developed; however, direct comparisons of the effects of these conditions have not occurred. We assessed a double factorial arrangement composed of six treatments (fresh or cryopreserved E. heros eggs from adults fed on natural or two artificial diets). We evaluated the biological characteristics and parasitism capacity of T. podisi produced from these treatments across seven temperatures. The thermal range between 21 and 30 °C resulted in satisfactory daily parasitism in all treatments tested, with an inverse relationship between temperature and female survival. The best parasitoid biological parameters were found between 21 and 27 °C, where all tested diets supported T. podisi development, with the best results from artificial diets. Fresh eggs and those frozen in liquid nitrogen and stored at -196 °C until use supported parasitoid development. These results suggest that the best method to mass rear T. podisi is to use artificial diets to rear E. heros and store eggs until needed, and then rear parasitoids at 24 °C.

Influence of artificial diets on biological characteristics and digestive enzymes of Coccinella septempunctata L.

In order to improve the rearing of Coccinella septempunctata L. (Coleoptera: Coccinellidae), nutrients such as shrimp, pollen, honey, and lard were added to the basic artificial diet, and the effects of the artificial diet on biological parameters and digestive enzymes were evaluated. The results show that beetles feeding on the supplemented diet exhibited pupation, emergence, fecundity, and hatching rates that were 102.69%, 125.02%, 162.33%, and 119.90% of those supplied with the basic diet, respectively. The addition of shrimp and pollen to the basal diet improved protease, trypsin, chymotrypsin, and aminopeptidase activity in larvae and female adults. The addition of lard improved lipase activity in female adults, and the addition of honey improved invertase activity in adults of both sexes. This study provides guidance for improving the nutritional benefits of ladybug artificial diets.

Efficacy of Artificial Diet on Economic Parameters of ERI Silkworm (Samia ricini Donovan)

The study was conducted to evaluate the efficiency of artificial diet on rearing performance of eri silkworm. The results revealed that in case of total larval duration (D:H) and matured larval weight (g) castor (control) fed larvae recorded least larval duration of 18 ± 0.8 D:H and highest matured larval weight of 8.190 g. Followed by treatment receiving artificial diet from fifth instar recorded 25 ± 0.9 D:H and 4.140 g. However, treatments receiving artificial diet from third and fourth instar were found to be inferior and on par with each other. With respect to cocoon parameters viz., cocoon weight, pupal weight, shell weight and shell ratio it was noticed that larvae receiving artificial diet from third and fourth instar didn’t spin cocoons. Effective Rate of Rearing (ERR %) indicated the poor performance of artificial diet and it needs better optimization for improve the performance of eri silkworm.

Temperature and Host Plant Impacts on the Development of Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae): Linear and Nonlinear Modeling

The tobacco cutworm, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae), is one of the most serious pests in field crops, vegetables, and ornamentals. Temperatures (15, 20, 25, 27, 30, 35, and 40 °C), host plants (soybean (Glycine max (L.)), maize (Zea mays L.), groundnut (Arachis hypogaea L.) and azuki bean (Vigna angularis (Willd.) Ohwi & H. Ohashi,), and the artificial diet-dependent developmental parameters and survival of S. litura were examined in this study. Stage-specific parameters such as threshold development temperature (LDT) and thermal constant (K) (Degree day (DD)) were determined by linear and nonlinear models (Sharpe-Schoolfield-Ikemoto), respectively. The total developmental time (egg-adult) decreased with increasing temperature on host plants and with an artificial diet. The total immature developmental time varied from 106.29, 107.57, 130.40, 111.82, and 103.66 days at 15 °C to 22.47, 21.25, 25.31, 18.30, and 22.50 days at 35 °C on soybean, maize, groundnut, azuki bean, and artificial diet, respectively. The LDT for the total immature completion was 7.50, 9.48, 11.44, 12.32, and 7.95 °C on soybean, maize, groundnut, azuki bean, and artificial diet, respectively. The K for the total immature completion was 587.88, 536.84, 517.45, 419.44, and 586.95 DD on soybean, maize, groundnut, azuki bean, and artificial diet, respectively. Temperature and host plant interaction also influenced the longevity and survival of adults. The findings of this study can be used to predict the number of generations, spring emergence, and population dynamics of S. litura. The nutrient content analysis of the host plants is discussed in terms of the developmental patterns of S. litura.

Effects of artificial diet breeding on intestinal microbial populations at the young stage of silkworm (Bombyx mori).

The silkworm (Bombyx mori) is an economically important insect and serves as a model organism for Lepidoptera. To investigate the effects of the intestinal microbial population on the growth and development of larvae fed an artificial diet (AD) during the young stages, we analyzed the characteristics of the intestinal microbial population using 16S rRNA gene sequencing technology. Our results revealed that the intestinal flora of the AD group tended to be simple by the 3rd-instar, which Lactobacillus accounting for 14.85% and leading to a decreased pH in the intestinal fluid. In contrast, the intestinal flora of silkworms in the mulberry leaf (ML) group showed continuous growth of diversity, with Proteobacteria accounting for 37.10%, Firmicutes accounting for 21.44%, and Actinobacteria accounting for 17.36%. Additionally, we detected the activity of intestinal digestive enzymes at different instars and found that the activity of digestive enzymes in the AD group increased by larval instar. Protease activity in the AD group was lower during the 1st- to 3rd-instars compared to the ML group, while α-amylase and lipase activities were significantly higher in the AD group during the 2nd- and 3rd-instar compared to the ML group. Furthermore, our experimental results indicated that changes in the intestinal population decreased the pH and affected the activity of proteases, which might contribute to the slower growth and development of larvae in the AD group. In summary, this study provides a reference for investigating the relationship between artificial diet and intestinal flora balance.

Effect of natural and artificial diets on the biological and reproductive characteristics of Cydia pomonella (Lepidoptera: Tortricidae)

Effect of natural and artificial diets on the biological and reproductive characteristics of Cydia pomonella (Lepidoptera: Tortricidae)

Feasibility of using fish visceral trash in a polyculture system for enhancing the growth performances of giant gourami (Osphronemus gouramy) and redclaw crayfish (Cherax quadricarinatus)

Giant gourami and redclaw crayfish are among popular freshwater organisms with significant potential for aquaculture due to their ease of cultivation, resistance to diseases, and high economic value. Therefore, this study aimed to evaluate the feasibility of using fish visceral trash (FVT) in artificial diets to enhance the growth performances of giant gourami (Osphronemus gouramy) and redclaw crayfish (Cherax quadricarinatus) in a polyculture system. Giant gourami juveniles (3.62±0.86 cm and 3.74±1.15 g) and redclaw crayfish (3.05±0.35 cm and 6.41±0.29 g) were divided into four groups and cultured in 100 L aquarium. Each group was fed twice daily with diets containing 0%, 10%, 20%, and 30% FVT/kg diet. Furthermore, growth performances were assessed in all samples at 14 days intervals over a period of eight weeks. The results showed that diets prepared with 30% FVT significantly affected the growth rate of giant gourami juveniles, while those comprising 20% enhanced the growth of redclaw crayfish. Total length, body weight, specific growth rate, and weight gain significantly increased in samples fed with the experimental compounded diets. Based on the observations, it is suggested that using FVT compounded diets tends to improve growth performances in a polyculture system.Keywords:Giant gouramiGrowth performanceRedclaw crayfishPolycultureTrash fish

Oxidative and genotoxic effect of piperazine on Galleria mellonella (Lepidoptera: Pyralidae) hemolymph.

Recently, there are many studies suggesting antibacterial, antifungal, and anthelmintic agents as alternative chemicals to insecticides. In this study, the oxidative and genotoxic effect of Piperazine, a clinically important hexahydropyrazine anthelmintic, on Galleria mellonella L. hemolymph tissue by adding artificial diet were investigated. Galleria mellonella larvae were reared until 7th larval stage in artificial diet containing 0.001, 0.01, 0.1, and 1 g piperazine per 100 g of diet. Using hemolymph collected from 7th-instar larvae, the amount of lipid peroxidation final product malondialdehyde (MDA), protein oxidation product protein carbonyl (PCO), and detoxification enzymes glutathione S-transferase (GST) and cytochrome P450 monooxygenase (cyt P450) activity, comet assay were measured. According to the results obtained, when the piperazine high concentrations tested with the control group were compared, statistically significant differences were found in MDA, PCO content, cyt P450, GST activity, and comet assay in the hemolymph of the insect. While MDA content was 0.01 ± 0.0021 nmol/mg protein in the control group, this amount increased approximately 2-fold at the highest concentration (0.0231 ± 0.0050 nmol/mg protein). On the other hand, when the control group and the highest piperazine concentration were compared in the GST and cyt P450 activity, it was determined that there was a statistically significant increase. We obtained similar results in comet assay and micronucleus formation data. This study showed that the tested piperazine concentrations caused significant changes in the detoxification capacity, oxidative stress, and genotoxic markers in the insect's hemolymph tissue.

Efficacy of an existing Ceratitis capitata (Diptera: Tephritidae) cold disinfestation treatment schedule for Ceratitis cosyra.

The efficacy of an existing cold disinfestation postharvest treatment targeting Ceratitis capitata (Wiedemann) in citrus was determined for the marula fly, Ceratitis cosyra (Walker). The cold tolerances of C. capitata and C. cosyra were first quantified in artificial diet at 3.5 °C at different exposure periods for up to 18 days. Ceratitis capitata was found to be more cold tolerant than C. cosyra. At 3.5 °C, the duration to achieve 99.9968% mortality was calculated to be 11.57 days for C. capitata and 9.10 days for C. cosyra. Under an existing C. capitata cold treatment schedule at 1 °C for 14 days, the conditions required for complete mortality of the third larval stage of C. cosyra in orange, Citrus sinensis (L.) Osbeck cv. Valencia, were then determined. No survivors of C. cosyra in oranges were recorded beyond 11 days of cold treatment at 1 °C. The efficacy of this C. capitata treatment for disinfestation of C. cosyra was thereafter confirmed in large scale trials in Valencia oranges. In the large-scale trial at the lowest mean temperature of 1.19 °C for 14 days, there were no survivors from a total of 85 490 treated C. cosyra third instars in oranges. Since C. capitata was shown to be more cold tolerant than C. cosyra and a large scale test demonstrated at least 99.9965% efficacy after 14 days at 1.19 °C, compared with the established effective C. capitata cold treatment of 14 days at 1.11 °C, cold disinfestation treatments for C. capitata should be at least equally effective against C. cosyra.

Cadmium exposure-triggered growth retardation in Hyphantria cunea larvae involves disturbances in food utilization and energy metabolism

Serious environmental pollution in the ecosystem makes phytophagous insects face a great risk of exposure to pollutants, especially heavy metals. This study aims to understand the effects of Cd exposure on the growth and development of Hyphantria cunea and to elucidate the mechanism of growth toxicity induced by Cd from the perspective of food utilization and energy metabolism. Our results showed that the larval basal growth data, growth index, fitness index, and standard growth index were significantly decreased after feeding on Cd-containing artificial diets. The Cd-treated larvae had significantly higher digestibility than the untreated larvae. However, the food consumption, efficiency of conversion of digested food, and efficiency of conversion of ingested food were significantly lower than those of untreated larvae. Eight key metabolites in the glycolysis pathway and six key metabolites in the tricarboxylic acid cycle pathway were significantly reduced in Cd-treated larvae. The mRNA expression levels of two regulatory genes (6-phosphofructokinase 1 and hexokinase-1) belonging to two key enzymes in the glycolysis pathway and four regulatory genes (isocitrate dehydrogenase-1, isocitrate dehydrogenase-3, citrate synthase, and oxoglutarate dehydrogenase) belonging to three key enzymes in the tricarboxylic acid cycle pathway were significantly lower in the Cd-treated group than in the control group. Furthermore, most fitness-related traits were significantly and positively correlated with food utilization (except approximate digestibility) or energy metabolism parameters. Taken together, Cd exposure-triggered growth retardation of H. cunea larvae is a consequence of disturbances in food utilization and energy metabolism, thereby emphasizing the toxicity of heavy metals.

An improved bioassay for the testing of entomopathogenic nematode virulence to the western corn rootworm (Diabrotica virgifera virgifera) (Coleoptera: Chrysomelidae): with focus on neonate insect assessments.

Bioassays involving newly hatched larval insects can be limited by the larvae's feeding state. Assays attempting to monitor mortality effects can be negatively affected by starvation effects on the larvae. Neonate western corn rootworms have significant reductions in viability if not provided food within 24 h post hatch. The recent development of an improved artificial diet for western corn rootworm larvae provides a new bioassay type for evaluating entomopathogenic nematodes that also makes the testing arena easy to observe. Here, we evaluated four species of entomopathogenic nematodes including Heterorhabditis bacteriophora Poinar, Steinernema carpocapsae (Weiser), Steinernema diaprepesi Nguyen & Duncan, and Steinernema rarum (de Doucet) against neonate western corn rootworm, Diabrotica virgifera virgifera LeConte, in 96-well plate diet bioassays. Nematode inoculation levels were 0, 15, 30, 60, and 120 nematodes per larva. Percentage mortality increased for each species as the rate of inoculation increased. Overall, H. bacteriophora and S. carpocapsae caused the greatest amount of larval mortality. The diet-based bioassays were shown to be an effective method for nematode exposure to insect pests. The assays provided adequate moisture to keep nematode from desiccating while also allowing freedom of movement around the arenas. Both rootworm larvae and nematodes were contained within the assay arenas. The addition of nematodes did not cause any significant deterioration of the diet within the three-day period of testing. Overall, the diet bioassays worked well as a measure of entomopathogenic nematode virulence.

Insecticidal potential of organic extracts of Calotropis procera to Spodoptera frugiperda

This study evaluated the toxic effects of organic extracts of Calotropis procera leaves on the survival, development, and reproduction of Spodoptera frugiperda. Solutions of crude methanol extract and hexane and methanol fractions of C. procera leaves were added at 1.15% and 2.14% concentrations to the artificial diet of S. frugiperda. The mortality and duration of larval and pupal phases, weights of female and male pupae, deformations of pupae and adults, the reduction of adults able to reproduce, pre-oviposition and oviposition periods, the number of postures per female, and the fecundity and fertility of S. frugiperda females were also evaluated. The extracts harmed the survival, development, and reproduction of S. frugiperda. The ingestion of extracts and fractions by caterpillars affected adults by decreasing the oviposition period, the number of postures, fecundity, and fertility. The crude MeOH extract at a 2.14% concentration harmed the evaluated parameters of the insect, except for pupal mortality, female pupae weight, and pre-oviposition period. The MeOH fraction at 2.14% caused a 50.0% mortality of caterpillars and 16.0% deformation in pupae and 33.0% in adults, reducing by 72.0% the population able to reproduce. The MeOH fraction at the 2.14% concentration caused 25.0% and 38.0% of pupal mortality and deformation, respectively. Calotropis procera has promising insecticidal properties for a biological insecticide, a convenient and sustainable strategy for protecting plants against S. frugiperda.

Xanthotoxin: An Aphicidal Coumarin From Ficus petiolaris against Melanaphis sacchari Zehntner (Hemiptera: Aphididae)

In this research, we evaluated the aphicidal effect of the ethanolic extract of stems and bark of Ficus petiolaris Kunth (Moraceae), in laboratory bioassays in an artificial diet against apterous adult females of Melanaphis sacchari Zehntner (Hemiptera: Aphididae). The extract was evaluated at different concentrations (500, 1,000, 1,500, 2,000, and 2,500 ppm), and the highest percentage of mortality (82%) was found at 2,500 ppm after 72 h. The positive control imidacloprid (Confial®) at 1% eliminated 100% of the aphids, and the negative control (artificial diet) only presented mortality of 4%. The chemical fractionation of the stem and bark extract of F. petiolaris yielded five fractions of FpR1-5, which were each evaluated at 250, 500, 750, and 1,000 ppm. FpR2 had the strongest aphicidal effect, with 89% mortality at 72 h at 1,000 ppm. The pure xanthotoxin compound extracted from this fraction was even more effective, with 91% aphid mortality after 72 h at 100 ppm. The lethal concentration (LC50) of xanthotoxin was 58.7 ppm (72 h). Our results indicate that the extract of F. petiolaris showed toxic activity against this aphid, and its xanthotoxin compound showed strong aphicidal activity at low concentrations.

A SIMPLE MASS REARING METHOD FOR BRINJAL SHOOT AND FRUIT BORER &lt;i&gt;LEUCINODES ORBONALIS&lt;/i&gt; GUENEE

A simple, effective and economically cheaper method for mass rearing of the brinjal shoot and fruit borer, Leucinodes orbonalis Guenee was developed using both modified artificial diet fortified with brinjal powder and natural diet - potato. A comparison of the various biological parameters such as egg incubation period, larval period, larval weight (12-day old), pre-pupal period, pupal period, pupal weight (1-day old), adult longevity, fecundity, oviposition period and sex ratio of different life stages of the test-insect were studied for both the food sources in this investigation. The larval and pupal periods on natural (potato) diet were 14.58± 1.51 and 6.7± 1.22 days respectively, with a mean fecundity of 165.55± 36.26 per female and a mean life cycle of 26.91days. The larval and pupal period of shoot and fruit borer reared on modified diet were 15.97± 1.24 and 6.25± 1.23 days respectively with a mean fecundity of 149.45± 25.83 per female and a mean life cycle of 25.26 days. Provision of a suitable site for adult mating and oviposition was attained by designing a separate mating chamber along with a suitable egg laying substratum. Thus, an easy and economical rearing technique could successfully support the rearing to six generations of target-insect in laboratory.

Antioxidant availability trades off with warning signals and toxin sequestration in the large milkweed bug (Oncopeltus fasciatus).

In some aposematic species the conspicuousness of an individual's warning signal and the concentration of its chemical defense are positively correlated. Several mechanisms have been proposed to explain this phenomenon, including resource allocation trade-offs where the same limiting resource is needed to produce both the warning signal and chemical defense. Here, the large milkweed bug (Oncopeltus fasciatus: Heteroptera, Lygaeinae) was used to test whether allocation of antioxidants, that can impart color, trade against their availability to prevent self-damage caused by toxin sequestration. We investigated if (i) the sequestration of cardenolides is associated with costs in the form of changes in oxidative state; and (ii) oxidative state can affect the capacity of individuals to produce warning signals. We reared milkweed bugs on artificial diets with increasing quantities of cardenolides and examined how this affected signal quality (brightness and chroma) across different instars. We then related the expression of warning colors to the quantity of sequestered cardenolides and indicators of oxidative state-oxidative lipid damage (malondialdehyde), and two antioxidants: total superoxide dismutase and total glutathione. Bugs that sequestered more cardenolides had significantly lower levels of the antioxidant glutathione, and bugs with less total glutathione had less luminant orange warning signals and reduced chroma of their black patches compared to bugs with more glutathione. Bugs that sequestered more cardenolides also had reduced red-green chroma of their black patches that was unrelated to oxidative state. Our results give tentative support for a physiological cost of sequestration in milkweed bugs and a mechanistic link between antioxidant availability, sequestration, and warning signals.

No physiological costs of dual sequestration of chemically different plant toxins in the milkweed bug Spilostethus saxatilis (Heteroptera: Lygaeidae)

Many herbivorous insects not only cope with plant toxins but also sequester them as a defense against predators and parasitoids. Sequestration is a product of the evolutionary arms race between plants and herbivorous insects and has been hypothesized to incur physiological costs due to specific adaptations required. Contradictory evidence about these costs exists for insects sequestering only one class of toxin, but very little is known about the physiological implications for species sequestering structurally different classes of compounds. Spilostethus saxatilis is a milkweed bug belonging to the cardenolide-sequestering heteropteran subfamily Lygaeinae (Heteroptera: Lygaeidae) that has shifted to the colchicine-containing plant Colchicum autumnale, a resource of chemically unrelated alkaloids. Using feeding-assays on artificial diet and chemical analysis, we assessed whether S. saxatilis is still able to sequester cardenolides apart from colchicine and related metabolites (colchicoids), and tested the effect of (1) either a natural cardenolide concentration (using ouabain as a model compound) or a natural colchicine concentration, (2) an increased concentration of both toxins, and (3) seeds of either Asclepias syriaca (cardenolides) or C. autumnale (colchicoids) on a set of life-history traits. For comparison, we assessed the same life-history traits in the milkweed bug Oncopeltus fasciatus exposed to cardenolides only. Although cardenolides and colchicoids have different physiological targets (Na+/K+-ATPase vs tubulin) and thus require different resistance traits, chronic exposure and sequestration of both isolated toxins caused no physiological costs such as reduced growth, increased mortality, lower fertility, or shorter adult life span in S. saxatilis. Indeed, an increased performance was observed in O. fasciatus and an according trend was found in S. saxatilis when feeding on isolated ouabain and isolated colchicine, respectively. Positive effects were even more pronounced when insects were provided with natural toxic seeds (i.e. C. autumnale for S. saxatilis and A. syriaca for O. fasciatus), especially in O. fasciatus. Our findings suggest, that S. saxatilis can sequester two chemically unrelated classes of plant compounds at a cost-free level, and that colchicoids may even play a beneficial role in terms of fertility.

An Efficient Rearing Method for the Fall Armyworm, &lt;i&gt;Spodoptera frugiperda&lt;/i&gt;, under Crowded Conditions

The fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), is an economically important polyphagous pest, which has recently expanded its distribution from the Americas to over 70 countries. Here, we established an efficient rearing method for S. frugiperda under crowded conditions. First, we clarified the larval density at which there is no decrease in survival rate due to cannibalism. Spodoptera frugiperda larvae were maintained on an artificial diet till pupation under five density conditions: 10, 30, 50, 70, and 100 individuals per container (5.4L, 26.8 cm × 20.7 cm × 9.8 cm). Survival rates at adult emergence were approximately 70% at density conditions of less than 50, and those in the container with paper towels as refuge were higher than those in the container without paper towels. In the second experiment, two individuals at different developmental stages from 5th instar to pupa were released in a container with or without food, and with or without paper towels, to clarify the developmental stages at which cannibalism occurs. Even when food was present, the individuals without access to refuge during the prepupa were attacked by the 6th instar larva. Our results suggested that conditions which include refuge and moderate density facilitate the rearing of S. frugiperda larvae under crowded conditions.