The effect of short-term food restriction on the metabolic cost of the acute phase response in the fish-eating Myotis (Myotis vivesi)

Food restriction affects the activation of the immune system although the metabolic cost associated with mounting such a response has rarely been examined except in model animals. Wild animals are constantly exposed to variations in the availability of food resources and they need to balance their energy budget to fight against pathogens. We examined the effect of food restriction in the fish eating Myotis (Myotis vivesi), a species of bat that experiences periods in which foraging is limited due to ambient conditions. We tested the hypothesis that acute food restriction (∼65% restriction for 1 night) would reduce the caloric response to lipopolysaccharidae (LPS) injection compared to bats fed ad libitum. We also measured a proxy for body temperature (T_skin) and expected reduced fever development when food intake was limited. Bats on the restricted diet had similar resting metabolic rate, total caloric cost and T_skin after the LPS challenge than when fed ad libitum. However, there was a delay in the metabolic and pyrogenic responses when bats were on the restricted diet. The effect of acute food restriction in delaying the hyperthermia development in fish eating Myotis might be of importance for its capacity to fight pathogens. Similar to other bats, the fish eating Myotis can fast for several consecutive days by entering torpor and future work is warranted to understand the effect of long periods of food restriction on bat immune response.     

Catch-up growth in Japanese quail (Coturnix Japonica): relationships with food intake, metabolic rate and sex

The effects of early environmental conditions can profoundly affect individual development and adult phenotype. In birds, limiting resources can affect growth as nestlings, but also fitness and survival as adults. Following periods of food restriction, individuals may accelerate development, undergoing a period of rapid “catch-up” growth, in an attempt to reach the appropriate size at adulthood. Previous studies of altricial birds have shown that catch-up growth can have negative consequences in adulthood, although this has not been explored in species with different developmental strategies. Here, we investigated the effects of resource limitation and the subsequent period of catch-up growth, on the morphological and metabolic phenotype of adult Japanese quail (Coturnix japonica), a species with a precocial developmental strategy. Because males and females differ in adult body size, we also test whether food restriction had sex-specific effects. Birds that underwent food restriction early in development had muscles of similar size and functional maturity, but lower adult body mass than controls. There was no evidence of sex-specific sensitivity of food restriction on adult body mass; however, there was evidence for body size. Females fed ad lib were larger than males fed ad lib, while females subjected to food restriction were of similar size to males. Adults that had previously experienced food restriction did not have an elevated metabolic rate, suggesting that in contrast to altricial nestlings, there was no metabolic carry-over effect of catch-up growth into adulthood. While Japanese quail can undergo accelerated growth after re-feeding, timing of food restriction may be important to adult size, particularly in females. However, greater developmental flexibility compared to altricial birds may contribute to the lack of metabolic carryover effects at adulthood.     

Resource limitation,predation risk and compensatory growth in a damselfly

Periods of poor nutrition during early development may have negative fitness consequences in subsequent periods of ontogeny. In insects, suppression of growth and developmental rate during the larval stage are likely to affect size and timing of maturity, which in turn may lead to reduced reproductive success or survivorship. In light of these costs, individuals may achieve compensatory growth via behavioural or physiological mechanisms following food limitation. In this study, we examined the effects of a temporary period of food restriction on subsequent growth and age and size at maturity in the larval damselfly Ischnura verticalis (Odonata: Coenagrionidae). We also asked whether this temporary period of reduced nutrition affected subsequent foraging behaviour under predation risk. I. verticalis larvae exposed to a temporary food shortage suffered from a reduced growth rate during this period relative to a control group that was fed ad libitum. However, increased growth rates later in development ensured that adult body size measurements (head and pronotum widths) did not differ between the treatments upon emergence. In contrast, adult dry mass did not catch up to that of the controls, indicating that the increased growth rates for size dimensions occur at the cost of similar gains in mass. Predators reduced foraging effort of larvae, but this reduction did not differ between control larvae and those previously exposed to poor nutrition.     

Energy Budget,Behavior and Leptin in Striped Hamsters Subjected to Food Restriction and Refeeding

Food restriction induces a loss of body mass that is often followed by rapid regaining of the lost weight when the restriction ends, consequently increasing a risk of development of obesity. To determine the physiological and behavioral mechanisms underlining the regaining, striped hamsters were restricted to 85% of initial food intake for 4 weeks and refed ad libitum for another 4 weeks. Changes in body mass, energy budget, activity, body composition and serum leptin level were measured. Body mass, body fat mass and serum leptin level significantly decreased in food-restricted hamsters, and increased when the restriction ended, showing a short “compensatory growth” rather than over-weight or obesity compared with ad libitum controls. During restriction, the time spent on activity increased significantly, which was opposite to the changes in serum leptin level. Food intake increased shortly during refeeding, which perhaps contributed to the rapid regaining of body mass. No correlation was observed between serum leptin and energy intake, while negative correlations were found in hamsters that were refed for 7 and 28 days. Exogenous leptin significantly decreased the time spent on activity during food restriction and attenuated the increase in food intake during refeeding. This suggests that low leptin in restricted animals may function as a starvation signal to induce an increase in activity behavior, and high leptin likely serves as a satiety signal to prevent activity during refeeding. Leptin may play a crucial role in controlling food intake when the restriction ends, and consequently preventing overweight.     

A 3-week feed restriction after weaning as an alternative to a medicated diet: effects on growth,health, carcass and meat traits of rabbits of two genotypes

Feed restriction after weaning is widely used in meat rabbit farms to promote health and reduce mortality, but this practice impacts negatively on rabbit growth and slaughter performance. This study compared a 3-week post-weaning feed restriction with ad libitum medicated feeding, evaluating effects on feed intake, growth, health, carcass and meat quality of rabbits of two genotypes: Italian White pure breed and Hycole hybrid×Italian White crossbred. A total of 512 rabbits at 36 days of age, of both sexes and two genotypes, were divided into four homogeneous groups assigned, from 36 to 57 days of age, to different feeding programmes (FP): restricted non-medicated (R-N), ad libitum non-medicated (L-N), restricted medicated (R-M) and ad libitum medicated (L-M). The diets were medicated with oxytetracycline (1540 mg/kg) and colistin sulphate (240 mg/kg). The restriction, performed by giving 70, 80 and 90 g/day of feed for the 1st, 2nd and 3rd week, was followed by ad libitum feeding in the successive 5 weeks, up to slaughter at 92 days of age. Restricted feeds were ingested at a level of 64% of the feed intake recorded in the ad libitum fed rabbits; it was significantly associated, regardless of medication and rabbit genotype, with a lower feed intake (−22 to −24 g dry matter/day) during the entire experiment, compensatory growth and a lower feed conversion ratio in the ad libitum period, and a lower final live weight (−150 g) than ad libitum feeding (P<0.001). During restriction, mortality was lower in the restricted rabbits (6.25%, 5.47% v. 12.5%, 14.8% for R-N, R-M, L-N and L-M; P<0.05), whereas in the ad libitum period mortality did not differ among the groups (9.23%, 9.90%, 11.0% and 4.59% for R-N, R-M, L-N and L-M). Dressing out percentage was not affected by FP or genotype; heavier carcasses were produced by rabbits fed ad libitum (+100 g; P<0.001) and crossbred rabbits (+122 g; P<0.001). Restriction did not alter meat quality, except for a tendency towards a higher cooking loss and less fat; crossbred meat was higher in L* (+1.3; P<0.01) and b* (+0.51; P<0.05) colour indexes and tenderness (−0.14 kg/cm²; P<0.05) than pure breed meat. Under the conditions of this study, a 3-week restricted feeding after weaning resulted to be a suitable alternative, also for high growth potential genotypes, to the antibiotics to preserve rabbit health. The production of lighter carcasses could be compensated partly by the lower feed conversion ratio showed by restricted rabbits.     

Food restriction followed by refeeding with a casein- or whey-based diet differentially affects the gut microbiota of pre-pubertal male rats

Researchers are gaining an increasing understanding of host–gut microbiota interactions, but studies of the role of gut microbiota in linear growth are scarce. The aim of this study was to investigate the effect of food restriction and refeeding with different diets on gut microbiota composition in fast-growing rats. Young male Sprague–Dawley rats were fed regular rat chow ad libitum (control group) or subjected to 40% food restriction for 36 days followed by continued restriction or ad libitum refeeding for 24 days. Three different diets were used for refeeding: regular vegetarian protein chow or chow in which the sole source of protein was casein or whey. In the control group, the composition of the microbiota remained stable. Food restriction for 60 days led to a significant change in the gut microbiota at the phylum level, with a reduction in the abundance of Firmicutes and an increase in Bacteroidetes and Proteobacteria. Rats refed with the vegetarian protein diet had a different microbiota composition than rats refed the casein- or whey-based diet. Similarities in the bacterial population were found between rats refed vegetarian protein or a whey-based diet and control rats, and between rats refed a casein-based diet and rats on continued restriction. There was a significant strong correlation between the gut microbiota and growth parameters: humerus length, epiphyseal growth plate height, and levels of insulin-like growth factor 1 and leptin. In conclusion, the type of protein in the diet significantly affects the gut microbiota and, thereby, may affect animal's health.     

High temperature slows down growth in tobacco hornworms (Manduca sexta larvae) under food restriction

When fed ad libitum (AL), ectothermic animals usually grow faster and have higher metabolic rate at higher ambient temperature. However, if food supply is limited, there is an energy tradeoff between growth and metabolism. Here we hypothesize that for ectothermic animals under food restriction (FR), high temperature will lead to a high metabolic rate, but growth will slow down to compensate for the high metabolism. We measure the rates of growth and metabolism of 4 cohorts of 5th instar hornworms (Manduca sexta larvae) reared at 2 levels of food supply (AL and FR) and 2 temperatures (20 and 30 °C). Our results show that, compared to the cohorts reared at 20 °C, the ones reared at 30 °C have high metabolic rates under both AL and FR conditions, but a high growth rate under AL and a low growth rate under FR, supporting this hypothesis.     

Maternal dopamine exposure provides offspring starvation resistance in Daphnia

The neurotransmitter dopamine has been shown to play an important role in modulating behavioral, morphological, and life history responses to food abundance. However, costs of expressing high dopamine levels remain poorly studied and are essential for understanding the evolution of the dopamine system. Negative maternal effects on offspring size from enhanced maternal dopamine levels have previously been documented in Daphnia. Here, we tested whether this translates into fitness costs in terms of lower starvation resistance in offspring. We exposed Daphnia magna mothers to aqueous dopamine (2.3 or 0 mg/L for the control) at two food levels (ad libitum vs. 30% ad libitum) and recorded a range of maternal life history traits. The longevity of their offspring was then quantified in the absence of food. In both control and dopamine treatments, mothers that experienced restricted food ration had lower somatic growth rates and higher age at maturation. Maternal food restriction also resulted in production of larger offspring that had a superior starvation resistance compared to ad libitum groups. However, although dopamine exposed mothers produced smaller offspring than controls at restricted food ration, these smaller offspring survived longer under starvation. Hence, maternal dopamine exposure provided an improved offspring starvation resistance. We discuss the relative importance of proximate and ultimate causes for why D. magna may not evolve toward higher endogenous dopamine levels despite the fitness benefits this appears to have.     

Modelling Effects of Temperature and Feeding Level on the Life Cycle of the Midge Chironomus Riparius: An Energy-Based Modelling Approach

Survival, growth, emergence and reproduction were monitored during the life cycle of Chironomus riparius in the laboratory at five temperatures from 15 to 27 °C and two feeding diets (ad libitum and limiting). Data were analysed using an energy-based model. Survival was not affected. In ad libitum conditions, growth rate increased linearly with temperature as a consequence of quicker food ingestion. Our model parameterised using the data in ad libitum conditions could account for the data obtained in food limited conditions. Reproduction was not influenced by temperature because the decrease of the duration of the period of energetic investment into reproduction compensated for the increase of the feeding rate when temperature increased. Based on these results, we then built a model describing life cycles in the field which may contribute to the field assessment of the consequences of global warming or pollution.     

Fatty acid oxidation and fatty acid synthesis in energy restricted rats

The importance of fat oxidation and fatty acid synthesis were examined in rats fed approximately one half their ad libitum food intake for a period of 13 days followed by 7 days of ad libitum feeding (refed rats). This study was undertaken because previous reports demonstrated that refed rats rapidly accumulated body fat. Our results confirmed this observation: refed rats accrued body fat and body weight at rates that were approximately 3 times higher than controls. Evidence for a period of increased metabolic efficiency was demonstrated by measuring the net energy requirement for maintenance over the refeeding period: refed rats had a reduced metabolic rate during the period of energy restriction (approximately 30% lower than control) and this persisted up to 2 days after the reintroduction of ad libitum feeding. The major factor responsible for the rapid fat gain was a depressed rate of fatty acid oxidation. Calculations of protein and carbohydrate intake over the refeeding period showed that the simplest explanation for the decrease in fatty acid oxidation is fat sparing. This is possible because of the large increase in dietary carbohydrate and protein intake during the refeeding period when metabolic rates are still depressed. The increased carbohydrate and protein may adequately compensate for the increasing energy requirements of the ER rats over the refeeding period affording rats the luxury of storing the excess dietary fat energy.     

Effects of larval density and food stress on life‐history traits of Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)

High population density and nutrition restriction can lead to phase variation in morphology and development, and subsequently induce changes in the reaction norms of adult flight in migrant insects. However, response of migratory propensity to such stress in Endopterygote insects, especially in several species of Lepidoptera, remains unclear. In this study, larval and adult developmental responses to crowding and food stress were investigated in the migratory moth, Cnaphalocrocis medinalis (Guenée). A high larval rearing density significantly reduced pupal mass, survival rate and female fecundity. Larvae developed rapidly under crowding conditions, and time to pupation was 2 days earlier than individuals reared alone. By contrast, short‐term starvation and associated compensatory growth prolonged larval duration by 3–4 days and pupal duration by 1–2 days. It also reduced the pupal mass, but showed no detectable effects on female reproductive performance. Both sexes had similar development strategies; however, females seemed to be more sensitive to crowding and food shortage than males. A positive effect was expected if such stress factors acted as cues that triggering a behavioural or physiological shift to a distinct migratory phase. To the contrary, we found no proof that crowding and starvation caused maturation delay in female reproductive development. All treatments did not significantly increase female pre‐oviposition period. Therefore, we concluded that life developmental responses to crowding and food shortage in this species were different. Adult migration propensity was not enhanced under such stress conditions during the larval phase.     

Food restriction affects circadian and photoperiodic response in black-headed munia (Lonchura malacca malacca)

We determined the effect of food restriction on locomotor activity pattern and gonadal growth on black-headed munia (Lonchura malacca malacca) exposed to12L:12D light–dark cycle. Two groups of male and female munia (n = 5 pairs each) were exposed to 12L:12D (L = 265 lux; D = 0.3 lux) for day 1–7. On day 8, group one received 4 h food (ZT 0–4; ZT 0 = Zeitgeber time, light onset) and group two received food ad libitum (ZT 0–24 h). Locomotor activity pattern and gonadal growth were recorded over the experiment period of one year. Birds were highly active in group one, with a high daily and total activity. However, activity behavior was fully synchronized with light onset in the group two, when food was available ad libitum. Gonadal maturation was initiated in both the groups but the maturation of testis and ovarian follicle was noticed only in group two (food ad libitum). It appears that food restriction affects the locomotor activity and gonadal growth in black-headed munia.     

Energetic challenges unmask the role of ovarian hormones in orchestrating ingestive and sex behaviors

Effects of ovarian hormones on sex and ingestive behavior are well studied, and yet, their role in diverting attention from food to sex has not been examined directly, possibly because these functions are masked under conditions of excessive food abundance typical of the laboratory. Female Syrian hamsters were either fed ad libitum or food-restricted to 75% of their ad libitum intake for 8 days and then tested every day of the estrous cycle for their preference for males versus food, food hoarding and food intake in an apparatus designed to mimic aspects of their natural habitat. The food-restricted, but not the fed females, varied significantly over the estrous cycle in appetitive behaviors, which included their preference for males versus food and in the amount of food hoarded, with low food hoarding and high male preference on the night of ovulation. In contrast, there were no significant differences between restricted and ad libitum-fed females in the consummatory behaviors, namely, food intake or lordosis duration. In ovariectomized females, estradiol plus progesterone treatment delayed food restriction-stimulated hoarding and hastened feeding-inhibited hoarding without affecting food intake or lordosis duration. In summary, energy restriction and the presence of males unmasked an effect that was obscured in the normal laboratory conditions characterized by isolation and an over abundance of readily available food. These results are consistent with the idea that ovarian hormones orchestrate appetites for food and sex to optimize reproductive success under fluctuating energetic conditions.     

Larval food limitation in butterflies: effects on adult resource allocation and fitness

Allocation of larval food resources affects adult morphology and fitness in holometabolous insects. Here we explore the effects on adult morphology and female fitness of larval semi-starvation in the butterfly Speyeria mormonia. Using a split-brood design, food intake was reduced by approximately half during the last half of the last larval instar. Body mass and forewing length of resulting adults were smaller than those of control animals. Feeding treatment significantly altered the allometric relationship between mass and wing length for females but not males, such that body mass increased more steeply with wing length in stressed insects as compared to control insects. This may result in changes in female flight performance and cost. With regard to adult life history traits, male feeding treatment or mating number had no effect on female fecundity or survival, in agreement with expectations for this species. Potential fecundity decreased with decreasing body mass and relative fat content, but there was no independent effect of larval feeding treatment. Realized fecundity decreased with decreasing adult survival, and was not affected by body mass or larval feeding treatment. Adult survival was lower in insects subjected to larval semi-starvation, with no effect of body mass. In contrast, previous laboratory studies on adult nectar restriction showed that adult survival was not affected by such stress, whereas fecundity was reduced in direct 11 proportion to the reduction of adult food. We thus see a direct impact of larval dietary restriction on survival, whereas fecundity is affected by adult dietary restriction, a pattern reminiscent of a survival/reproduction trade-off, but across a developmental boundary. The data, in combination with previous work, thus provide a picture of the intra-specific response of a suite of traits to ecological stress.     

A semi-artificial rearing system for the specialist predatory ladybird Cryptolaemus montrouzieri

In the present study a semi-artificial rearing system for the Australian ladybird Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae), a specialist predator of mealybugs, was developed. In a first step, a rearing system using eggs of the Mediterranean flour moth Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) as a food and synthetic polyester wadding as an oviposition substrate was compared with a natural rearing system using the citrus mealybug, Planococcus citri (Risso) (Hemiptera: Pseudococcidae), as to its effects on the predator’s developmental and reproductive parameters. In a second series of experiments the performance of C. montrouzieri on bee pollen or on a mixture of E. kuehniella eggs and bee pollen was assessed. E. kuehniella eggs proved to be a suitable food to support larval development of the predator. Ladybird larvae reared on flour moth eggs developed two days faster and weighed approximately 10 % more than their counterparts reared on mealybugs. Despite a prolongation of the preoviposition period with ca. eight days and a decrease in egg hatch by about 10 %, C. montrouzieri females fed moth eggs accepted the synthetic wadding as an oviposition substrate and deposited the same number of eggs their counterparts maintained on mealybugs. A mixture of E. kuehniella eggs with pollen yielded similar developmental and reproductive rates as E. kuehniella eggs alone, but a diet of bee pollen alone was not adequate for the predator. Our findings indicate the potential of a rearing system using E. kuehniella eggs as a factitious food and synthetic wadding as an artificial oviposition substrate for the mass production of C. montrouzieri.     

Metabolic consequences of overlapping food restriction and cell‐mediated immune response in a long‐distance migratory shorebird,the little ringed plover Charadrius dubius

Investment in immunity is commonly viewed as an energetically costly activity in birds. Although several studies have focused on the energy cost of mounting an immune response and its concomitant physiological trade‐offs, nothing is known about the metabolic adjustments experienced by immunochallenged birds under resource limitation, or about the basal metabolism cost of mounting cell‐mediated immune (CMI) responses in bird species other than non‐migratory passerines. Here we measured the basal metabolic rate (BMR), inflammatory response, and body mass in ad libitum fed and food‐restricted little ringed plovers Charadrius dubius challenged with phytohemagglutinin (PHA) in order to assess the energy cost, the strength, and the time course of the CMI response in a long‐distance migratory bird in different nutritional states. We found that ad libitum birds injected with PHA significantly increased both mass‐independent BMR and inflammatory response, whereas birds with an induced food restriction‐immune response overlap experienced a mass‐independent BMR downregulation and decreased inflammatory response relative to ad libitum birds. We suggest that both the BMR downregulation and the diminished inflammatory response observed in birds facing such an overlap could be energy‐saving mechanisms to maintain the body mass above a critical level and maximize fitness.     

Haematological and serum biochemical responses of chickens to hydric stress

Dehydration can be extremely damaging to the performance and welfare of indigenous chickens. The effect of water restriction on haematological and biochemical parameters was compared in Naked Neck (NNK) and Ovambo (OVB) chickens. A total of 54 8-week-old pullets each of NNK and OVB chickens with an initial average weight of 641 ± 10 g/bird were randomly assigned to three water intake treatments with three replications, each having six birds. The water restriction treatments were ad libitum, 70% and 40% of ad libitum intake. Nine experimental pens with a floor space of 3.3 m² per strain were used. Feed was provided ad libitum. Packed cell volume (PCV), erythrocyte count (RBC), mean corpuscular volume (MCV) and total leucocyte count (WBC), and biochemical parameters (uric acid (UA)), creatinine (CREAT), total protein (TP), albumin (ALB), globulin (GLOB), triglyceride (TGA), total cholesterol (TC), high- (HDLC) and low- (LDLC) density lipoprotein cholesterol and activity of alanine transaminase (ALT), alkaline phosphatase (ALP) and aspartate transaminase (AST) were determined from blood collected after 60 days of water restriction. PCV was higher (P < 0.05) in NNK than OVB chickens offered water ad libitum, but similar in birds offered 70% and 40% of ad libitum. There were no differences in RBC and MCV values between strains, but MCV was higher in birds on 40% than 70% of ad libitum water intake, irrespective of strain. Naked neck chickens had higher (P < 0.05) WBC values than OVB at 40% restriction level, but lower WBC than OVB at 70% water restriction level. UA, CREAT, TGA, TC, LDLC, TP and GLOB increased (P < 0.05) with each increment in water restriction, but the increase in CREAT and TC was more pronounced in OVB than NNK chickens. The opposite was observed for UA. ALT activity indicated that liver function was not affected by water restriction. It was concluded that the two strains can withstand up to 40% of ad libitum water restriction, but NNK tolerated water stress better than OVB chickens.     

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.     

Dietary influence of cardenolides on larval growth and development of the milkweed bug Oncopeltus fasciatus

Larvae of the milkweed bug Oncopeltus fasciatus were reared on the seeds of eight different species of milkweed (Asclepias), representing a wide range of cardenolide concentrations in the diet. There were few significant differences in larval developmental period, wet body weight of teneral adults, dry weight of adults, and pronotal width of adults reared on the different diets. However, the data indicate no significant correlations between cardenolide content, and body weight or size of the adult insects.There was no evidence in this study of a physiological cost or adverse effect on the larval growth and development of insects which sequestered and stored differing quantities of cardenolides. Instead, the data support a recently-proposed model of cardenolide sequestration which may be energy-independent.The validity of evidence supporting a physiological cost hypothesis for sequestration of cardenolides by the monarch butterfly is discussed in the light of these findings.