Nutrition as a neglected factor in insect herbivore susceptibility to Bt toxins.

The widespread global adoption of Bt crops elevates concerns about the evolution of Bt resistance in insect pest species. Current insecticide resistance management (IRM) strategies focus solely on genetic variation as a causal factor in the evolution of resistance, but ignore the role that environmental factors, such as nutrition, may play. In this opinion paper, we discuss the benefits that insect herbivores gain from consuming foods with protein-carbohydrate content that matches their self-selected protein-carbohydrate intake, and show that even within monocultures there is amply opportunity for insect herbivores to regulate their macronutrient intake. Next we review new data that show that dietary protein and carbohydrates can: firstly, have predictably strong effects on the survival and performance of caterpillars challenged with Bt toxins, and secondly, mediate plasticity in susceptibility to Cry1Ac, which can account for large differences in LC50 values. Nutrition-Bt interactions such as these have important implications for IRM, particularly given that diet-incorporated Bt bioassays commonly use artificial diets that vary substantially from their self-selected optimal diets, which likely results in underestimates of resistance in the field. Failing to bioassay larvae on ecologically-relevant diets can seriously confound the results of Bt resistance monitoring bioassays and undermine our ability to detect resistance in the field.

Sterol metabolic constraints as a factor contributing to the maintenance of diet mixing in grasshoppers (Orthoptera: Acrididae).

Sterols are essential nutrients for all arthropods, including grasshoppers, but metabolic constraints may limit which sterols can support normal growth and development. In the firsts part of this study, a comparative experiment, which included five different species of grasshoppers (Orthoptera: Acrididae) representing three separate taxonomic groups, was performed to determine how widespread sterol metabolic constraints are within the Acrididae. Grasshoppers were reared on artificial diets containing sterols that differed in the position of double bonds within the sterol structure, and various life history traits were measured. Sterols with double bonds at position 7, within the sterol nucleus, and/or at position 22, on the cholestane side chain, failed to support development to the adult stage for any of the five species. In addition, grasshoppers reared on sterols with these configurations often had extended developmental times and reduced growth rates in the first and second stadium compared with grasshoppers reared on sitosterol or cholesterol diets. In the second half of this study, we examined how mixtures of suitable and unsuitable sterols influenced survival, growth, and development. Artificial foods containing mixtures of suitable and unsuitable sterols were fed to the highly polyphagous grasshopper Schistocerca americana. Results suggest that survival and performance of this grasshopper suffer as the concentration of unsuitable sterols increases and as the ratio of suitable to unsuitable sterols in the diet decreases. We review the literature to document variation in plant sterol profiles and propose that constraints on sterol metabolism may contribute to the maintenance of diet mixing in the Acrididae.

Effects of diet on titratable acid-base excretion in grasshoppers.

Despite the potential for diet to affect organismal acid-base status, especially in herbivores, little is known about the effects of diet on acid-base loading and excretion. We tested the effects of diet on acid-base loading and excretion in grasshoppers by (a) comparing the fecal acid-base content of 15 grasshopper species collected from the field and (b) comparing fecal acid-base excretion rates of Schistocerca americana grasshoppers fed vegetable diets that differed in their ashed and raw acid-base contents. The field experiments indicated that grass-feeding species excrete fairly neutral fecal pellets, while forb/mixed-feeding species vary widely in their fecal acid-base contents. In the laboratory experiment, acid-base excretion rates were positively correlated with dietary ashed base intake rates but were not correlated with the acid-base content of raw, unashed diet or feeding rate. These experiments suggest that some diets could strongly challenge the acid-base homeostasis of herbivores; in some grasshoppers, dietary acid-base loads could produce certainly lethal 1-unit changes in average body pH within 6 h if they were not excreted.

The nutritional significance of sterol metabolic constraints in the generalist grasshopper Schistocerca americana.

Sterols are essential nutrients for grasshoppers, as well as all other insects, but metabolic constraints can limit which phytosterols support normal growth and development. In the current study, the generalist grasshopper Schistocerca americana was used to address two questions related to grasshopper sterol nutrition: (1) how does sterol quantity influence growth and survival, and (2) how do mixtures of suitable and unsuitable sterols at different concentrations influence growth and survival? Results from the first experiment indicated that this grasshopper species had a minimum sterol requirement of 0.05% dry weight; as sterol quantity increased above this concentration, however, survival and performance were not enhanced. Results from the second experiment revealed two novel aspects of sterol nutrition in grasshoppers: (1) when suitable sterols were limiting, most individuals could not use unsuitable sterols to meet the minimum sterol requirement (i.e. no sparing occurred), and (2) above a certain threshold, unsuitable sterols were deleterious even when suitable sterols were present at a concentration that alone permits normal growth and development. We discuss these physiological findings in terms of how sterol metabolic constraints in grasshoppers might influence foraging.

Post-ingestive feedbacks and associative learning regulate the intake of unsuitable sterols in a generalist grasshopper.

Behavioural studies of the grasshopper Schistocerca americana were undertaken to identify the mechanisms that regulate the intake of dietary sterols. In the first experiment, grasshoppers were allowed to feed on spinach, a plant containing only unsuitable sterols; immediately after this first meal, a suitable or unsuitable sterol was injected into the haemolymph. Grasshoppers injected with unsuitable sterols had second meals on spinach that were significantly shorter than those of grasshoppers injected with suitable sterols, indicating that unsuitable dietary sterols are detected post-ingestively. In the second experiment, grasshoppers were fed food containing only unsuitable sterols and were then presented with glass-fibre discs containing different concentrations of a suitable sterol or sucrose only (the control). The results suggest that grasshoppers do not use a direct feedback operating on mouthpart chemoreceptors to regulate their intake of suitable sterols. In the third experiment, grasshoppers were presented with artificial diets containing different sterols and flavours, and feeding was observed over a sequence of meals. The results from both the first and last experiments suggest a role for associative learning in regulating the intake of unsuitable sterols.

Phytosterol metabolism and absorption in the generalist grasshopper, Schistocerca americana (Orthoptera: Acrididae).

A series of experiments, using GLC, RP-HPLC, and GC-MS techniques, were performed to examine the metabolic fate and absorption of different dietary sterols in the grasshopper Schistocerca americana. In the first experiment, grasshoppers were reared on diets containing different sterols presented singly. Cholesterol was the dominant tissue sterol recovered from cholesterol and "soybean sitosterol" fed grasshoppers but among the grasshoppers fed diets with stigmasterol and spinach sterols (both unsuitable for growth and development), the amount of cholesterol recovered was not different from that of newly hatched grasshoppers. In the second experiment, grasshoppers were given diets containing mixtures of soybean sitosterol and stigmasterol and the metabolic fate of these dietary sterols was recorded. Results from this experiment suggest that the presence of an unsuitable dietary sterol does not interfere with cholesterol production from sitosterol. They also demonstrate that large quantities of unmetabolized dietary sterols with C-24 ethyl groups are recovered from grasshoppers fed diets containing stigmasterol. Finally, tissue sterol profiles of grasshoppers with and without their midguts were compared. Results suggest that the midgut is the major tissue where unmetabolized dietary sterols accumulate. How these sterol metabolic constraints impact development and survival is discussed as well as the impact they might have on grasshopper feeding behavior. Arch.

A comparison of nutritional regulation in solitarious- and gregarious-phase nymphs of the desert locust Schistocerca gregaria.

Nutritional regulatory responses were compared for the cryptic 'solitarious' and the conspicuously coloured, aggregating 'gregarious' phases of the desert locust Schistocerca gregaria. The desert locust has the genetic potential to exist in either phase, changing between them within a lifetime and epigenetically across generations. Our aim was to compare final-instar nymphs of the two phases with respect to key nutritional variables, including (i) points of regulated intake (the 'intake target') for protein and carbohydrate, (ii) the nature of trade-offs between over-eating nutrients in excess and under-eating those in deficit when fed nutritionally unbalanced foods, (iii) diet-related patterns of nutrient utilisation, and (iv) the performance consequences of eating nutritionally unbalanced diets. When provided with pairs of nutritionally unbalanced but complementary foods, both phases regulated their intake of protein and carbohydrate to a similar point. However, when confined to foods that were of unbalanced protein to carbohydrate ratio, gregarious nymphs ate more than solitarious insects. Both phases regulated protein growth, but gregarious insects did so to a lower adult body protein content and converted ingested protein to growth less efficiently. When fed a food high in carbohydrate and low in protein, gregarious nymphs deposited more body lipid and survived less well than did solitarious insects. Solitarious nymphs developed more quickly than gregarious nymphs except on the two most extremely unbalanced diets, on which development time was similar. The results are discussed with respect to the different nutritional ecologies of the two phases and used to develop the hypothesis that animals have evolved to trade-off the cost of eating excess of a nutritionally unbalanced diet against the probability of encountering foods of complementary composition in the future.

A geometric analysis of nutrient regulation in the generalist caterpillar Spodoptera littoralis (Boisduval).

Nutritional regulatory responses of the generalist caterpillar Spodoptera littoralis (Boisduval) were explored, in choice and no-choice experiments, using the Geometric Framework. In the choice experiment, newly moulted final instar larvae were provided with one of three protein-biased foods (PB-food: p35:c7, p28:c5.6 or p21:c4.2) and one of three equal protein-carbohydrate ratio foods (ER-food: p21:c21, p16.8:c16.8 or p12.6:c12.6). On five of the nine treatments, caterpillars independently regulated protein and carbohydrate intake to a mixture of 57 and 43%, respectively. However, when the concentration of the ER-food decreased and that of the paired PB-food increased, caterpillars progressively abandoned regulation and ate more of the PB-food. Despite these regulatory differences, performance (survivorship, growth and development) was similar across all nine treatments. In the no-choice experiment, caterpillars were given one of five foods (p35:c7, p28:c14, p21:c21, p14:c28 and p7:c35). Results indicated that caterpillars moved to a point in protein-carbohydrate space that was consistent with the Equal Distance Rule of compromise, a pattern previously predicted for generalist feeders. The insects on the two extreme foods, p35:c7 and p7:c35, showed reduced pupal mass and longer development, respectively. There was also strong evidence for post-ingestive regulation of nutrient utilisation, notably for protein.