Changes in midgut and brain proteins in Morimus funereus larvae depending on nutritive substrate

The response of Morimus funereus larvae to total starvation and refeeding with qualitatively different nutritive substrates (artificial diets supplemented with yeast as a source of B complex vitamins or with a digestibility reducer-tannic acid) was examined in this paper. Refeeding resulted in a compensatory increase of larval growth. Feeding and refeeding with qualitatively different nutritive substrates affected both quality and quantity of midgut and brain proteins. The observed differences suggest the possible switching of enzyme isoforms in M. funereus midgut and changes in synthesis/secretion of neurohormones, depending on food presence and its nutritional value.     

Plant silicon effects on insect feeding dynamics are influenced by plant nitrogen availability

Although there is growing evidence that silicon (Si)‐based plant defenses effectively reduce both the palatability and digestibility of leaves, and thus impact nutrient assimilation by insect herbivores, much less is known about how this is affected by extrinsic and intrinsic factors. For example, do herbivores exhibit compensatory feeding on poor‐quality diets with Si or are Si defenses less effective in agroecosystems where high N availability increases plant quality? To investigate the interactive effects of N and Si on insect feeding, we conducted insect performance and compensatory feeding bioassays using maize, Zea mays L. (Poaceae), and the true armyworm, Pseudeletia unipuncta Haworth (Lepidoptera: Noctuidae). In the performance assay, the addition of Si alone resulted in increased larval mortality compared with the controls, likely because early instars with poorly developed mandibles could not feed effectively. However, larvae fed on plants treated with both Si and N survived better than on plants treated with Si only, although pupal mass did not differ between treatments. In our compensatory assay, Si addition reduced maize consumption, but increased both armyworm approximate digestibility and N assimilation efficiency, suggesting that enhanced post‐ingestion feeding physiology, rather than compensatory food intake, could have accounted for the lack of Si effects on pupal weight. Overall, our results demonstrate that, similar to other chemical and mechanical defenses, the effectiveness of plant Si defense is influenced by plant nutrient status and consumer compensatory ability.     

Compensatory responses of Samea multiplicalis larvae when fed leaves of different fertilization levels of the aquatic weed Pistia stratiotes

Compensatory responses of caterpillars fed low quality food include increased consumption and utilization of essential nutrients. Information about an insect's responses to nutritional challenges from their host plants could benefit weed biological control efforts in the selection and establishment of new agents. The target weed, Pistia stratiotes L. (Araceae) is a floating aquatic plant that has relatively low nitrogen levels which are further diluted with high water content. Efforts to establish the insect Spodoptera pectinicornis (Hampson) (Lepidoptera: Noctuidae) for biological control of P. stratiotes could benefit by examining the nutritional responses of a similar widely established lepidopteran species, Samea multiplicalis (Guenèe) (Lepidoptera: Pyralidae). Larvae of this species were fed leaves of P. stratiotes plants that had been fertilized (NPK) at high and low rates. The leaves of the fertilized plants had a 4.3-fold increase in nitrogen (dry weight) and a 1.6-fold increase in water content. The results suggest that no compensatory increases occurred in larvae fed leaves from the low fertilized plants as no changes were found in fresh mass consumption or nitrogen utilization efficiency. Consequently, development time from second-third instars to pupation was delayed about 3 days compared with larvae fed the high nitrogen leaves. Furthermore, consumption of nitrogen was only 30% and its accumulation into larval tissues was only 60% compared with the larvae fed the high fertilized leaves. The resulting larvae had both a final biomass and a growth rate that were reduced by 40%. Regardless of plant fertilizer level, the larvae fed at a rate 5–10 times greater than that of similar lepidopteran species consuming either low or high quality diets, suggesting that the S. multiplicalis larvae may be functioning at their biological limit for ingesting food.     

Using nutritional indices to study LOX3-dependent insect resistance

Induced resistance to biotic attackers is thought to be mediated by responses elicited by jasmonic acid (JA), a subset of which are lipoxygenase 3 (LOX3) dependent. To understand the importance of LOX3-mediated insect resistance, we analysed the performance of Manduca sexta larvae on wild-type (WT) and on isogenic Nicotiana attenuata plants silenced in NaLOX3 expression and JA signalling, and we used Waldbauer nutritional indices to measure the pre- and post-ingestive effects. LOX3-mediated defenses reduced larval growth, consumption and frass production. These defenses reduced how efficiently late-instar larvae converted digested food to body mass (ECD). In contrast, LOX3-mediated defenses decreased approximate digestibility (AD) in early instar larvae without affecting the ECD and total food consumption. Larvae of all instars feeding on defended WT plants behaviourally compensate for their reduced body mass by consuming more food per unit of body mass gain. We suggest that larvae feeding on plants silenced in NaLOX3 expression (as-lox) initially increase their AD, which in turn enables them to consume more food in the later stages and consequently, to increase their ECD and efficiency of conversion of ingested food (ECI). We conclude that N. attenuata's oxylipin-mediated defenses are important for resisting attack from M. sexta larvae, and that Waldbauer nutritional assays reveal behavioural and physiological counter responses of insects to these plant defenses.     

Coupling diet quality and Bowman-Birk and Kunitz-type soybean proteinase inhibitor effectiveness to Diatraea saccharalis development and mortality

We used bioassays to investigate the effect of Bowman‐Birk and Kunitz‐type soybean proteinase inhibitors on two artificial diets (diets 1 and 2) which are commonly used to feed laboratory colonies of larvae of the moth Diatraea saccharalis, monitoring food intake and utilization, and larval development and mortality. Diet 1 was less nutritious, with a low protein content and reduced mineral and essential amino acid (e.g., cysteine, lysine, and methionine) content, while diet 2 was richer and more complete. When proteinase inhibitors were incorporated into the artificial diets, the effects on larval development were significantly greater for those larvae fed diet 1, with the chronic ingestion of proteinase inhibitors reducing the level of trypsin‐like activity in the midgut of larvae fed this diet. Larvae fed diet 2 also showed a reduced level of tryptic activity in the midgut, but this was less marked than for diet 1. These results indicate that despite their inhibitory effect on midgut enzymes, the effectiveness of proteinase inhibitors is directly dependent on the quality of the diet. The different effects seen on insect biology when proteinase inhibitors are added to rich or poor diets suggests that the role of anti‐nutritional proteins in the control of insects might not be adequately addressed by bioassays based on the incorporation of inhibitors into artificial diets.     

Patterns and mechanisms of growth of fifth-instar Manduca sexta caterpillars following exposure to low- or high-protein food during early instars.

For many insect herbivores, variation in protein availability is a pervasive part of the environment. I explore how variable protein availability affects growth rates of fifth-instar Manduca sexta caterpillars and how growth is related to behavior and physiology. Groups of larvae were reared on low- or high-protein artificial diets (5.9% and 17.7% casein by dry weight, respectively) and then transferred in the fifth instar to the same or opposite diet. During or after the 24-h period following transfer, I measured growth rate, consumption rate, growth efficiency, midgut proteolytic activity, and masses of midgut contents and tissues. Fifth-instar caterpillars reared in earlier instars on high-protein diet grew about 20% more rapidly over 24 h than did caterpillars reared on low-protein diet. This growth pattern appears to be caused by differences in consumption and growth efficiency: caterpillars reared on high protein consumed more food, and used it more efficiently, than did caterpillars reared on low-protein diet. Over the short term (24 h), in contrast, fifth instars that received low-protein diet grew as rapidly as caterpillars that received high-protein diet. Increased (compensatory) consumption appears to be the primary mechanism by which caterpillars consuming low-protein food maintained growth rates.     

Modes of action of fraxinellone against the tobacco budworm, Heliothis virescens

Fraxinellone significantly reduced the relative growth rate, food consumption rate as well as the efficiency of conversion of ingested food into biomass of Heliothis virescens when incorporated into artificial diets at concentrations of 4.31 × 10⁻⁵ mol/L and above. After being fed with fraxinellone-treated diets for 24 h, the larval midguts of H. virescens possess significantly lower activities of α-amylase and non-specific proteases and higher activities of cytochrome P450s. In vitro , the compound did not inhibit the activities of α-amylase and non-specific proteases extracted from the larval midguts. Clear evidence of post-ingestive toxicity of fraxinellone to midgut cells was observed under an electron microscope. The modes of action of the compound against insects were discussed.     

Developmental responses to variable diet composition in a butterfly: the role of nitrogen,carbohydrates and genotype

Understanding the evolution of herbivore–plant interactions requires detailed information on proximate responses to relevant dietary variability and genetic variance, if any, associated with these responses. We measured the behavioral and developmental responses of Pieris rapae larvae to variation in the nitrogen (N) and carbohydrate (CARB) content of chemically‐defined diets, using differences in the average responses of sibling groups to estimate underlying genetic variance. Larval P. rapae responded to dietary reductions in both N and CARB with increased feeding, but these responses were inadequate to compensate for dietary N deficiencies within the range of N found in host plants. As a result, larvae on reduced N diets exhibited lower relative growth rates and longer development times, whereas larvae on reduced CARB diets maintained normal developmental trajectories. We also report evidence for genetic variation underlying (a) compensatory feeding responses to CARB and N availability, (b) N‐driven variation in growth rate and (c) CARB‐driven variation in larval duration. Our results highlight N availability as a key factor in the growth and development of this herbivorous butterfly, with CARB availability being less constraining and sufficiently addressed by changes to larval consumption rate. Furthermore, our study reveals standing genetic variance associated with larval responses to macronutrient availability, suggesting continued potential for herbivore–plant co‐evolution in this system despite a putative history of strong directional selection.     

Effects of poor food quality on copepod growth are dose dependent and non‐reversible

Herbivores regularly face imbalanced diets on a variety of timescales. They respond to such diets with reductions in growth and reproduction. The effect of exposure time to poor quality diets and recovery potential in herbivores has yet not been intensively studied. In order to investigate the response of herbivores growth and nutritional condition to phosphorus limited algal food, we fed the copepod Acartia tonsa with the autotrophic flagellate Rhodomonas salina, which was cultured towards high and low phosphorus content. To test the effect of the duration of the exposure to low and high P food on copepod growth, we also switched a subset of the copepods from high P to low P food for two to 12 days before putting them back on their original diet. Phosphorus limited prey clearly reduced copepod nutritional condition, expressed as their RNA:DNA ratio and consequently their growth rates. Several days after re‐feeding on high P algae, the copepods were able to reach growth rates comparable to the control groups, but they were in no case able to over shoot the growth rates of the control groups. A dose of two days feeding on low P algae resulted in slightly less than one stage growth reduction compared to copepods exclusively reared on high P algae. Our results show that phosphorus limited food drastically reduces secondary production, and no compensatory growth occurs in the investigated copepod species. Hence, even short term exposure of herbivores to poor food has a lasting effect on secondary production.     

Increased temperature reduces herbivore host‐plant quality

Globally increasing temperatures may strongly affect insect herbivore performance, as their growth and development is directly linked to ambient temperature as well as host‐plant quality. In contrast to direct effects of temperature on herbivores, indirect effects mediated via thermal effects on host‐plant quality are only poorly understood, despite having the potential to substantially impact performance and thereby to alter responses to the changing climatic conditions. We here use a full‐factorial design to explore the direct (larvae were reared at 17 °C or 25 °C) and indirect effects (host plants were reared at 17 °C or 25 °C) of temperature on larval growth and life‐history traits in the temperate‐zone butterfly Pieris napi. Direct temperature effects reflected the common pattern of prolonged development and increased body mass at lower temperatures. At the higher temperature, efficiency of converting food into body matter was much reduced being accompanied by an increased food intake, suggesting compensatory feeding. Indirect temperature effects were apparent as reduced body mass, longer development time, an increased food intake, and a reduced efficiency of converting food into body matter in larvae feeding on plants grown at the higher temperature, thus indicating poor host‐plant quality. The effects of host‐plant quality were more pronounced at the higher temperature, at which compensatory feeding was much less efficient. Our results highlight that temperature‐mediated changes in host‐plant quality are a significant, but largely overlooked source of variation in herbivore performance. Such effects may exaggerate negative effects of global warming, which should be considered when trying to forecast species' responses to climate change.     

Crowding-triggered phenotypic responses alleviate consequences of crowding inEpirrita autumnata (Lep., Geometridae)

Summary CrowdedEpirrita larvae had shorter larval periods than, and similar pupal masses to, their solitary siblings when reared on low quality diets. When fed on high quality diets, pupal masses of crowded larvae were lower than in singletons, and there was no difference in larval period. Because changes in food availability (absolute shortage, induced resistance in foliage) are caused by high larval densities in the field, crowding-triggered phenotypic changes may helpEpirrita to overcome detrimental consequences of high larval density. Pupal period was longer in crowded larvae than in singletons and crowded adults emerged later than their solitary siblings. Eggs of late emerging moths eclosed late in the ensuing spring, which coincides with delayed leaf flush in the year after defoliation. The reason for the faster growth of crowded individuals on poor diets was higher intake albeit less thorough processing of food in crowded, but not in solitary, larvae. On good diets solitary individuals tended to consume more than crowded larvae but there was no difference in processing. Predicted differences of host plant use between stealthy and opportunistic types of herbivores (sensu Rhoades 1985) were generally found between solitary and aggregated larvae on poor but not on good diets. The group response could not be explained by benefits to the group although the assumptions of Wilson's model of group selection were satisfied.     

Larval nutrition affects lipid storage and growth, but not protein or carbohydrate storage in newly eclosed adults of the grasshopper Schistocerca americana

Nitrogen availability from dietary protein can have profound effects on the physiology and evolutionary ecology of insect herbivores. While many studies consider the effects of nutrition on consumption and gross body composition of protein and other important nutrients, few consider partitioning to storage for future use. I used chemically defined artificial diets to quantitatively manipulate the amount of dietary carbohydrates and proteins available to growing larvae of the grasshopper Schistocerca americana to determine how larval nutrient availability affects growth and all three classes of stored nutrients (proteins, lipids, and carbohydrates) carried over from larval feeding into adulthood. Larvae on poor diets increased consumption, but could not compensate for diet quality, eclosing small and containing no significant nutrient stores at adulthood. Individuals fed intermediate to high nutrient content diets as larvae were significantly larger and contained a significantly greater proportion of lipid stores at adult eclosion, but not protein or carbohydrate stores than individuals fed low nutrient content diets. This suggests that larvally derived lipid stores may be more important to adult fitness than carbohydrate or protein stores. This result is contrary to previous studies performed on the role of larval nutrition and allocation to protein stores, and this difference is likely due to variation in the relative availability of protein in adult diets across species.     

Seasonal variation in leaf characteristics and food selectionby larval noctuids on an evergreen Mediterranean shrub

Despite year round availability of foliage, abundance of generalist noctuid larvae (Lepidoptera: Noctuidae) in evergreen-dominated Mediterranean forests has a narrow, distinct spring peak. This restricted larval period has been suggested to result in part from avoidance of the nutritionally poor mature foliage, and preference for nutritionally superior spring-produced young leaves. This study examines this hypothesis by (i) documenting differences in nutritional characteristics between expanding (April) and mature (June) young leaves of the evergreen Mediterranean shrub Daphne laureola L. (Thymelaeaceae), and (ii) experimentally studying the feeding preferences of noctuid larvae for young leaves, old leaves (≥ 1 yr old), and developing fruits of this species in one south-eastern Spanish locality. Young leaves of D. laureola declined in nutrient concentration and specific dry mass from April to June. The responses of noctuid larvae, in terms of both relative preference and total consumption, to this seasonal variation in chemical and physical features of young leaves were also investigated. When noctuid larvae were simultaneously offered young leaves, old leaves and developing fruits, they exhibited similar preferences for young leaves and developing fruits, and rejected old leaves developed during the previous year. Noctuid larvae did not modify their consumption of young leaves relative to old leaves and developing fruits in response to seasonal changes. Food selection patterns exhibited by D. laureola noctuid herbivores, notably the rejection of old leaves in favour of young ones, are consistent with the hypothesis relating restricted larval periods of these generalist consumers with the low food value of the previous season leaves of evergreen Mediterranean plants.     

Red foliage color reliably indicates low host quality and increased metabolic load for development of an herbivorous insect

Plant chemical defense and coevolved detoxification mechanisms in specialized herbivorous insects are fundamental in determining many insect–plant interactions. For example, Brassicale plants protect themselves from herbivory by producing glucosinolates, but these secondary metabolites are effectively detoxified by larvae of Pierid butterflies. Nevertheless, not all Brassicales are equally preferred by these specialist herbivores. Female Pieris butterflies avoid laying eggs on anthocyanin-rich red foliage, suggesting red color is a visual cue affecting oviposition behavior. In this study, we reared P. brassicae larvae on green and red cabbage leaves, to determine whether foliage color reliably indicates host plant quality. We did not find a difference in survival rates or maximal larval body mass in the two food treatments. However, larvae feeding on red cabbage leaves exhibited significantly lower growth rates and longer durations of larval development. Interestingly, this longer development was coupled with a higher consumption rate of dry food matter. The lower ratio of body mass gain to food consumption in larvae feeding on red cabbage leaves was coupled with significantly higher (ca. 10 %) larval metabolic rates. This suggests that development on red foliage may incur an increased metabolic load associated with detoxification of secondary plant metabolites. Energy and oxygen allocation to detoxification could come at the expense of growth and thus compromise larval fitness as a result of extended development. From an evolutionary perspective, red foliage color may serve as an honest defensive cue, as it reliably indicates the plant’s low quality as a substrate for larval development.     

Seasonal allocation of defense investment in Ilex opaca Aiton and constraints on a specialist leafminer

Summary Phytomyza ilicicola (Diptera: Agromyzidae), a univoltine specialist leafminer, is one of the few insect herbivores of American holly. Adult emergence is closely synchronized with leaf flush in spring, and females make numerous feeding punctures on and oviposit in new leaves. Larvae hatch in late May and June, but their feeding period and development are prolonged so that more than 80% of the mine enlargement occurs from January until March of the following year. We propose that this unusual life cycle reflects adaptation to constraints imposed by seasonal and age-related changes in chemical and structural defenses, and in nutritional quality of holly foliage. As holly leaves age, there is a shift in allocation of defense investment away from allelochemicals, including phenolic compounds and saponins, toward leaf sclerophylly, spinose teeth, and low foliar nitrogen and water. Rapid increases in leaf toughness and decreases in nutritional quality limit availability of leaf tissues for adult feeding and oviposition to a two-to threeweek phenological window during leaf flush. Mature holly foliage is a nutritionally poor resource by nearly all criteria known to affect food quality for herbivores. This may be the main reason for the prolonged larval development of P. ilicicola. Alternatively, winter feeding and pupation in spring may be adaptations which help to ensure synchrony of adult emergence with leaf flush. Repeated puncturing by female P. ilicicola does not render leaves more suitable for larvae, nor is it a means by which females sample leaf exudate to assess leaf quality prior to oviposition. Rather, leaf puncturing occurs mostly on leaves that are relatively high in soluble nitrogen, and is apparently a means by which females obtain protein and sugars prior to and during oviposition.The investigation reported in this paper (No. 85-7-8-208) is in connection with a project of the Kentucky Agricultural Experiment Station and is published with the approval of the Director     

Variation in temperature and dietary nitrogen affect performance of the gypsy moth (Lymantria dispar L.)

Abstract. . The independent and interactive effects of temperature and dietary nitrogen content on performance of the gypsy moth (Lymantria dispar L.) were examined. In long-term feeding trials, larvae were reared from egg hatch to pupation on low (1.5%) and high (3.7% dry weight) nitrogen diets, under three temperature regimes. Short-term feeding trials with fourth instars and the same treatments were conducted in order to calculate nutritional indices.
Higher temperatures did not influence larval survival and marginally increased final pupal weights, but strongly decreased long-term development rates. They also accelerated short-term growth and consumption rates, and tended to improve food processing efficiencies. High concentrations of dietary nitrogen increased survival rates and final pupal weights markedly, but decreased long-term development rates only marginally. A high content of dietary nitrogen also accelerated short-term development and growth rates, reduced consumption rates, and improved food digestibility. Insects responded to low nitrogen-content diets primarily by eating faster, rather than by altering efficiency of nitrogen use. In the short-term feeding trials, thermal regime and dietary nitrogen interacted to influence growth rates, overall food processing efficiencies and nitrogen consumption rates. No interactive effects were observed in long-term studies.
This research demonstrates that small changes in thermal regime and ecologically relevant variation in dietary nitrogen content can strongly affect gypsy moth performance. Moreover, various performance parameters are differentially sensitive to the direct and interactive effects of temperature and diet.     

Bryophyte use by an insect herbivore: does the crane‐fly Tipula montana select food to maximise growth?

1. Growth rates of immature invertebrates can be of profound importance to subsequent reproductive success in the adult stage. If diet selection in herbivores is a principal means of maximising growth and rate of development, individuals should select food of the highest quality. 2. If disparities are found between performance and preference, these may be the result of individuals balancing nutritional gain against other demands on their behaviour, such as a need for shelter. The responses of the crane‐fly herbivore Tipula montana Curtis (Diptera: Tipulidae) to a range of moss genera from an upland environment were investigated to determine whether diet selection was determined by food quality alone, as measured by its effect on larval growth and survival. 3. Larvae were reared individually on single moss genus diets, and their growth, development, and survival were measured. Each of the mosses supported growth and development although the mean weights achieved differed by a factor of two between food genera. In two‐choice preference experiments, individuals broadly chose bryophyte foods as expected from their quality. One notable exception was the moss Pleurozium schreberi, which gave the best growth performance but was among the least preferred. 4. The two methods of assessing preference gave different results: observations of larvae showed relatively greater selection for dense mosses than faecal pellet analysis. A strong dietary preference for an angiosperm, the sedge Carex bigelowii, over all mosses supported the interpretation that crane‐fly larvae benefit from bryophytes as a refuge, and that this factor may override dietary quality.     

Impact of dietary allelochemicals on gypsy moth (Lymantria dispar) caterpillars: importance of midgut alkalinity

Midgut pH of gypsy moth larvae was depressed artificially with buffered diet to examine the impact of alkalinity on the caterpillars' ability to tolerate a dietary polyphenol and a quinone. A 2x3 factorial design was used, with 2 levels of succinate buffer and 3 dietary amendments (tannic acid, juglone, or control). Development was monitored during the third and fourth instars, with consumption, food passage rates, midgut pH, and midgut redox potential (Eh) measured in the fourth instar. Diet buffering successfully depressed midgut pH to hypothetically suboptimal acidic levels without reductions in survivorship, but it did reduce larval growth and impede development. Buffering dramatically reduced survivorship of fourth instar larvae eating diets containing tannic acid or juglone. Growth increased on unbuffered diet amended with tannic acid, but not with juglone. Caterpillars passed food through the gut more slowly when feeding on buffered tannic acid diet or on unbuffered juglone diet. These results indicate that maintenance of midgut alkalinity is critical to tolerance of dietary tannic acid and juglone, and that these allelochemicals have very different activities in the caterpillar gut.     

Putting the insect into the birch–insect interaction

Leaf maturation in mountain birch (Betula pubescens ssp. czerepanovii) is characterized by rapid shifts in the types of dominant phenolics: from carbon-economic flavonoids aglycons in flushing leaves, via hydrolysable tannins and flavonoid glycosides, to carbon-rich proanthocyanidins (condensed tannins) in mature foliage. This shift accords with the suggested trade-offs between carbon allocation to plant defense and growth, but may also relate to the simultaneous decline in nutritive leaf traits, such as water, proteins and sugars, which potentially limit insect growth. To elucidate how birch leaf quality translates into insect growth, I introduce a simple model that takes into account defensive compounds but also acknowledges insect demand for nutritive compounds. The effects of defensive compounds on insect growth depend strongly on background variation in nutritive leaf traits: compensatory feeding on low nutritive diets increases the intake of defensive compounds, and the availability of growth-limiting nutritive compounds may modify the effects of defenses. The ratio of consumption to larval growth (both in dry mass) increases very rapidly with leaf maturation: from 2.9 to 9.8 over 2 weeks in June-July, and to 15 by August. High concentrations in mature birch leaves of "quantitative" defenses, such as proanthocyanidins (15-20% of dry mass), presumably prevent further consumption. If the same compounds had also protected half-grown leaves (which supported the same larval growth with only one third of the dry matter consumption of older leaves), the same intake of proanthocyanidins would have demanded improbably high concentrations (close to 50%) in young leaves. The model thus suggests an adaptive explanation for the high levels of "quantitative" defenses, such as proanthocyanidins, in low-nutritive but not in high-nutritive leaves because of the behavioral responses of insect feeding to leaf nutritive levels.