Effects of ambient temperature on glucose tolerance and insulin sensitivity test outcomes in normal and obese C57 male mice.

Mice are commonly used as animal models to study human metabolic diseases, but experiments are typically performed at room temperature, which is far below their thermoneutral zone and is associated with elevated heart rate, food intake, and energy expenditure. We set out to study how ambient temperature affects glucose tolerance and insulin sensitivity in control and obese male mice. Adult male C57BL/6J mice were housed at room temperature (23°C) for 6 weeks and fed either control or high fat diet. They were then fasted for 6 h before glucose or insulin tolerance tests were performed at 15, 20, 25, or 30°C. To ensure that behavioral thermoregulation did not counterbalance the afflicted ambient temperatures, oxygen consumption was determined on mice with the same thermoregulatory opportunities as during the tests. Decreasing ambient temperatures increased oxygen consumption and body mass loss during fasting in both groups. Mice fed high fat diet had improved glucose tolerance at 30°C and increased levels of fasting insulin followed by successive decrease of fasting glucose. However, differences between control and high-fat diet mice were present at all temperatures. Ambient temperature did not affect glucose tolerance in control group and insulin tolerance in either of the groups. Ambient temperature affects glucose metabolism in mice and this effect is phenotype specific.

Nutrient-specific compensatory feeding in a mammalian carnivore, the mink, Neovison vison.

Balancing of macronutrient intake has only recently been demonstrated in predators. In particular, the ability to regulate carbohydrate intake is little studied in obligate carnivores, as carbohydrate is present at very low concentrations in prey animal tissue. In the present study, we determined whether American mink (Neovison vison) would compensate for dietary nutritional imbalances by foraging for complementary macronutrients (protein, lipid and carbohydrate) when subsequently given a dietary choice. We used three food pairings, within which two macronutrients differed relative to each other (high v. low concentration), while the third was kept at a constant level. The mink were first restricted to a single nutritionally imbalanced food for 7 d and then given a free choice to feed from the same food or a nutritionally complementary food for three consecutive days. When restricted to nutritionally imbalanced foods, the mink were willing to overingest protein only to a certain level ('ceiling'). When subsequently given a choice, the mink compensated for the period of nutritional imbalance by selecting the nutritionally complementary food in the food choice pairing. Notably, this rebalancing occurred for all the three macronutrients, including carbohydrate, which is particularly interesting as carbohydrate is not a major macronutrient for obligate carnivores in nature. However, there was also a ceiling to carbohydrate intake, as has been demonstrated previously in domestic cats. The results of the present study show that mink regulate their intake of all the three macronutrients within limits imposed by ceilings on protein and carbohydrate intake and that they will compensate for a period of nutritional imbalance by subsequently selecting nutritionally complementary foods.

Balancing of specific nutrients and subsequent growth and body composition in the slug Arion lusitanicus.

Feeding generalists typically occupy broad ecological niches and so are potentially pre-adapted to a range of novel food objects. In northern Europe, the slug Arion lusitanicus has spread rapidly as an invasive species and a serious horticultural and agricultural pest. We used nutritional geometry to analyze nutrient balancing capabilities and consequences for performance in A. lusitanicus when provided with one of three nutritionally fixed diets or when given dietary choice. The slugs over-ingested high amounts of the most abundant nutrient in order to get more of the limited nutrient. However, they regulated protein intake more tightly than carbohydrate intake resulting in a very high food intake when fed a protein-poor diet. Growth and body composition were highly affected by the nutrient balance of their diet. When given the choice to feed from two nutritionally different diets, the slugs selected an intake balance of protein and carbohydrate with sufficient precision to maximize growth. Nutrient utilization efficiency increased with increasing deficiency of the specific nutrient in the diet. Ingested carbohydrate was more efficiently stored as lipid in slugs fed more carbohydrate-poor diets, and ingested nitrogen was more efficiently incorporated into slug bodies in slugs fed more protein-poor diets. Our experiments suggest that the evolved behavioral and physiological regulatory capacities of A. lusitanicus may explain some of the success that this slug experiences as an invasive species. We furthermore propose that invasive species might be more dependent on high protein availability in the environment than non-invasive species.

Optimal foraging for specific nutrients in predatory beetles.

Evolutionary theory predicts that animals should forage to maximize their fitness, which in predators is traditionally assumed equivalent to maximizing energy intake rather than balancing the intake of specific nutrients. We restricted female predatory ground beetles (Anchomenus dorsalis) to one of a range of diets varying in lipid and protein content, and showed that total egg production peaked at a target intake of both nutrients. Other beetles given a choice to feed from two diets differing only in protein and lipid composition selectively ingested nutrient combinations at this target intake. When restricted to nutritionally imbalanced diets, beetles balanced the over- and under-ingestion of lipid and protein around a nutrient composition that maximized egg production under those constrained circumstances. Selective foraging for specific nutrients in this predator thus maximizes its reproductive performance. Our findings have implications for predator foraging behaviour and in the structuring of ecological communities.

Prey nutrient composition has different effects on Pardosa wolf spiders with dissimilar life histories.

The nutritional composition of prey is known to influence predator life histories, but how the life history strategies of predators affect their susceptibility to nutrient imbalance is less investigated. We used two wolf spider species with different life histories as model predators: Pardosa amentata, which have a fixed annual life cycle, and Pardosa prativaga, which reproduce later and can extend development across 2 years. We fed juvenile spiders of the two species ad libitum diets of one of six Drosophila melanogaster fly types varying in lipid:protein composition during three instars, from the start of the second instar until the fifth instar moult. We then tested for interactions between predator species and prey nutrient composition on several life history parameters. P. amentata completed the three instars faster and grew larger carapaces and heavier body masses than P. prativaga, but the two species responded differently to variation in prey lipid:protein ratio. Duration of the instars increased when feeding on protein-poor prey in P. amentata, but was unaffected by diet in P. prativaga. Likewise, the effect of diet on body composition was more pronounced in P. amentata than in P. prativaga. Prey nutrient composition thus affected the two species differently. During macronutrient imbalance P. amentata appear to prioritize high growth rates while experiencing highly variable body compositions, whereas P. prativaga maintain more constant body compositions and have slower growth. These can be seen as different consequences of a fixed annual and a plastic annual-biennial life cycle.

Dietary protein content affects evolution for body size, body fat and viability in Drosophila melanogaster.

The ability to use different food sources is likely to be under strong selection if organisms are faced with natural variation in macro-nutrient (protein, carbohydrate and lipid) availabilities. Here, we use experimental evolution to study how variable dietary protein content affects adult body composition and developmental success in Drosophila melanogaster. We reared flies on either a standard diet or a protein-enriched diet for 17 generations before testing them on both diet types. Flies from lines selected on protein-rich diet produced phenotypes with higher total body mass and relative lipid content when compared with those selected on a standard diet, irrespective of which of the two diets they were tested on. However, selection on protein-rich diet incurred a cost as flies reared on this diet had markedly lower developmental success in terms of egg-to-adult viability on both medium types, suggesting a possible trade-off between the traits investigated.

Metabolic consequences of feeding and fasting on nutritionally different diets in the wolf spider Pardosa prativaga.

We investigated whether spiders fed lipid-rich rather than protein-rich prey elevate metabolism to avoid carrying excessive lipid deposits, or whether they store ingested lipids as a buffer against possible future starvation. We fed wolf spiders (Pardosa prativaga) prey of different lipid:protein compositions and measured the metabolic rate of spiders using closed respirometry during feeding and fasting. After a 16-day feeding period, spider lipid:protein composition was significantly affected by the lipid:protein composition of their prey. Feeding caused a large and fast increase in metabolism. The cost of feeding and digestion was estimated to average 21% of the ingested energy irrespective of diet. We found no difference in basal metabolic rate between dietary treatments. During starvation V 02 and V(CO)2decreased gradually, and the larger lipid stores in spiders fed lipid-rich prey appeared to extend survival of these spiders under starvation compared to spiders fed protein-rich prey. The results show that these spiders do not adjust metabolism in order to maintain a constant body composition when prey nutrient composition varies. Instead, lipids are stored efficiently and help to prepare the spiders for the long periods of food deprivation that may occur as a consequence of their opportunistic feeding strategy.

Protein and carbohydrate composition of larval food affects tolerance to thermal stress and desiccation in adult Drosophila melanogaster.

Larval nutrition may affect a range of different life history traits as well as responses to environmental stress in adult insects. Here we test whether raising larvae of fruit flies, Drosophila melanogaster, on two different nutritional regimes affects resistance to cold, heat and desiccation as well as egg production and egg-to-adult viability. We raised larvae on a carbohydrate-enriched and a protein-enriched growth medium. We found that flies developed on the high protein medium had increased heat and desiccation tolerance compared to flies developed on the carbohydrate-enriched medium. In contrast, flies developed on the carbohydrate-enriched growth medium recovered faster from chill coma stress compared to flies developed on a protein-enriched medium. We also found gender differences in stress tolerance, with female flies being more tolerant to chill coma, heat knockdown and desiccation stress compared to males. Egg production was highest in females that had developed on the protein-enriched medium. However, there was a sex-specific effect of nutrition on egg-to-adult viability, with higher viability for males developing on the sucrose-enriched medium, while female survival was highest when developing on the protein-enriched medium. Our study indicates that larval nutrition has a strong impact on the ability to cope with stress, and that the optimal nutrient composition varies with the type of stress.

Dietary ratio of protein to carbohydrate induces plastic responses in the gastrointestinal tract of mice.

Some vertebrates change the size of their digestive system in response to quantity and fibre content of ingested food, but the effects of dietary nutrients on gut structure remain poorly understood. Here we investigate how the protein to carbohydrate ratio of diets affects the mass of the gastrointestinal tract in mice. We fed 6-week-old male mice one of five isocaloric diets differing only in protein to carbohydrate ratio (the "no-choice" treatments), while a further four treatment groups received nutritionally complementary food pairings from which they could self-select a diet (the "choice" treatments). After 32 days, we measured the resulting dry mass of stomachs, intestines, caeca and colons. In the no-choice treatments, the stomachs were heavier in the mice fed diets containing more protein and less carbohydrate, indicating that larger stomachs may be needed for efficient digestion of the protein-rich food. In contrast, intestines, caeca and colons were heavier when diets contained more carbohydrates and less protein. This response may function to increase the digestive rate of carbohydrates when the dietary content of this macronutrient increases, but it may also indicate a compensatory response to increase amino acid uptake from a protein-deficient food. Mice in the choice treatments self-selected a diet with a protein to carbohydrate ratio of 0.46, and had gut dimensions similar to the expectation derived from no-choice treatments for this diet composition. Our results provide an example of plasticity in the differential allocation of resources to organ function, which is triggered by variation in resource quality.

Nutrient balance affects foraging behaviour of a trap-building predator.

Predator foraging may be affected by previous prey capture, but it is unknown how nutrient balance affects foraging behaviour. Here, we use a trap-building predator to test whether nutrients from previous prey captures affect foraging behaviour. We fed orb-weaving spiders (Zygiella x-notata) prey flies of different nutrient composition and in different amounts during their first instar and measured the subsequent frequency of web building and aspects of web architecture. We found that both the likelihood of web building and the number of radii in the web were affected by prey nutrient composition while prey availability affected capture area and mesh height. Our results show that both the balance of nutrients in captured prey and the previous capture rate may affect future foraging behaviour of predators.

Speed over efficiency: locusts select body temperatures that favour growth rate over efficient nutrient utilization.

Ectotherms have evolved preferences for particular body temperatures, but the nutritional and life-history consequences of such temperature preferences are not well understood. We measured thermal preferences in Locusta migratoria (migratory locusts) and used a multi-factorial experimental design to investigate relationships between growth/development and macronutrient utilization (conversion of ingesta to body mass) as a function of temperature. A range of macronutrient intake values for insects at 26, 32 and 38 degrees C was achieved by offering individuals high-protein diets, high-carbohydrate diets or a choice between both. Locusts placed in a thermal gradient selected temperatures near 38 degrees C, maximizing rates of weight gain; however, this enhanced growth rate came at the cost of poor protein and carbohydrate utilization. Protein and carbohydrate were equally digested across temperature treatments, but once digested both macronutrients were converted to growth most efficiently at the intermediate temperature (32 degrees C). Body temperature preference thus yielded maximal growth rates at the expense of efficient nutrient utilization.

The life of a dead ant: the expression of an adaptive extended phenotype.

Specialized parasites are expected to express complex adaptations to their hosts. Manipulation of host behavior is such an adaptation. We studied the fungus Ophiocordyceps unilateralis, a locally specialized parasite of arboreal Camponotus leonardi ants. Ant-infecting Ophiocordyceps are known to make hosts bite onto vegetation before killing them. We show that this represents a fine-tuned fungal adaptation: an extended phenotype. Dead ants were found under leaves, attached by their mandibles, on the northern side of saplings approximately 25 cm above the soil, where temperature and humidity conditions were optimal for fungal growth. Experimental relocation confirmed that parasite fitness was lower outside this manipulative zone. Host resources were rapidly colonized and further secured by extensive internal structuring. Nutritional composition analysis indicated that such structuring allows the parasite to produce a large fruiting body for spore production. Our findings suggest that the osmotrophic lifestyle of fungi may have facilitated novel exploitation strategies.

Protein-leverage in mice: the geometry of macronutrient balancing and consequences for fat deposition.

OBJECTIVE: The Protein-Leverage Hypothesis proposes that humans regulate their intake of macronutrients and that protein intake is prioritized over fat and carbohydrate intake, causing excess energy ingestion when diets contain low %protein. Here we test in a model animal, the mouse: (i) the extent to which intakes of protein and carbohydrate are regulated; (ii) if protein intake has priority over carbohydrates so that unbalanced foods low in %protein leads to increased energy intake; and (iii) how such variations in energy intake are converted into growth and storage. METHODS AND PROCEDURES: We fed mice one of five isocaloric foods having different protein to carbohydrate composition, or a combination of two of these foods (N = 15). Nutrient intake and corresponding growth in lean body mass and lipid mass were measured. Data were analyzed using a geometric approach for analyzing intake of multiple nutrients. RESULTS: (i) Mice fed different combinations of complementary foods regulated their intake of protein and carbohydrate toward a relatively well-defined intake target. (ii) When mice were offered diets with fixed protein to carbohydrate ratio, they regulated the intake of protein more strongly than carbohydrate. This protein-leverage resulted in higher energy consumption when diets had lower %protein and led to increased lipid storage in mice fed the diet containing the lowest %protein. DISCUSSION: Although the protein-leverage in mice was less than what has been proposed for humans, energy intakes were clearly higher on diets containing low %protein. This result indicates that tight protein regulation can be responsible for excess energy ingestion and higher fat deposition when the diet contains low %protein.

Nutrient-specific compensation following diapause in a predator: implications for intraguild predation.

In recent years it has become clear that intraguild predation (where predators feed on other predators) has important consequences for food webs, and yet very little is known about its nutritional or functional bases. In the most detailed study of the nutritional basis of foraging by a predator to date, we used geometrical analysis to test the ability of the generalist invertebrate predatory beetle, Agonum dorsale (Carabidae), to forage selectively for lipid and protein over a 10-day period following emergence from winter diapause, and we measured associated changes in body lipid and nitrogen content. Over the first 48 hours, beetles that were offered two nutritionally imbalanced but complementary foods self-selected a diet high in lipids, and thereafter the proportion of protein in the selected diet increased. Beetles confined to a single food with excess lipid (higher lipid:protein ratio than the self-selected diet) regulated intake to meet lipid requirements, while suffering a shortfall of protein. Those given diets with a lower lipid:protein ratio than the self-selected diet showed a progressive tendency across the 10-day experiment to over-ingest protein, thereby reducing the lipid deficit in their diet. Body composition changed markedly during the experiment, with the lipid content of the self-selecting insects increasing over the first 48 hours from 14% to 46% by dry mass, and thereafter remaining stable. We discuss some implications of our results for the understanding of intraguild predation.

Nutritional value of cannibalism and the role of starvation and nutrient imbalance for cannibalistic tendencies in a generalist predator.

1. Cannibalism is considered an adaptive foraging strategy for animals of various trophic positions, including carnivores. However, previous studies on wolf spiders have questioned the high nutritional value of cannibalism. We therefore analysed two different aspects of nutritional quality of conspecifics in the wolf spider Pardosaprativaga: their value for survival, growth and development; and the growth efficiency of feeding on conspecifics. We also measured the propensity for cannibalistic attacks and the consumption rate of conspecifics in an experiment where hunger level and nutrient balance were manipulated. In all experiments, cannibalism was compared with predation on fruit flies as control prey. 2. The growth experiment gave ambiguous results regarding the nutritional quality of conspecifics. Spiders on pure cannibalistic diets split into two distinct groups, one performing much better and the other much worse than spiders on fruit fly diets. We discuss the possibility that the population is dimorphic in its cannibalistic propensity, with the latter group of individuals showing a high level of inhibition against cannibalistic attacks in spite of a high nutritional value of cannibalism. 3. The food utilization experiment confirmed the high nutritional quality of conspecifics, as cannibalistic spiders had the same growth rate as spiders fed insect prey in spite of a much lower consumption rate. 4. Inhibition against cannibalistic attacks was demonstrated in medium-sized juveniles: only half of the spiders attacked a prescribed victim of 50% the size of their opponents, and the latency for those that did attack was more than half an hour, compared with a few minutes for spiders fed fruit flies. 5. Nutrient-imbalanced spiders utilized an alternative insect diet less efficiently than balanced spiders, whereas no difference was present in efficiency of utilizing conspecifics. This result indicates that spiders can remedy at least part of a nutrient imbalance through cannibalism. 6. As spiders can escape nutritional imbalance as well as restore energy reserves through cannibalism, we predicted both nutrient imbalance and hunger to stimulate cannibalism. This prediction was confirmed only with respect to hunger. Nutrient-imbalanced spiders had reduced cannibalistic consumption, perhaps due to lowered predatory aggressiveness as a result of bad condition.

Nutrient-specific foraging in invertebrate predators.

Many herbivores and omnivores adjust their food selection behavior to regulate the intake of multiple nutrients. Carnivores, however, are generally assumed to optimize the rate of prey capture rather than select prey according to nutrient composition. We showed experimentally that invertebrate predators can forage selectively for protein and lipids to redress specific nutritional imbalances. This selection can take place at different stages of prey handling: The predator may select among foods of different nutritional composition, eat more of a prey if it is rich in nutrients that the predator is deficient in, or extract specific nutrients from a single prey item.

Nutrient composition of the prey's diet affects growth and survivorship of a generalist predator.

It is well known that secondary chemicals produced at one trophic level may affect organisms at subsequent levels of the food chain. Effects of nutrient supplements may also propagate through trophic levels, but the mechanisms here are less clear. We tested the hypothesis that predators can be affected by the nutrient composition of the prey's food. Wolf spider (Pardosa amentata) hatchlings were raised ad libitum on fruit flies (Drosophila melanogaster) that were cultured in poor basic medium with additions of different nutrients. These additions strongly affected the performance of the spiders. Growth rates increased when additions consisted of 19 different amino acids or fatty acids+cholesterol or commercial dogfood. Survival increased in spiders reared on fruit flies from cultures containing 19 amino acids or methionine or dogfood. The addition of dogfood increased spider growth and survival more significantly than the addition of any single nutrient group alone. Adult female flies from the dogfood culture were significantly heavier than females from the basic culture. The nutrients added to the fruit fly media were thus able to create biological effects at both the second and the third trophic levels. To test whether nutrients passed to the predators through the gut content of the prey, we included a treatment where the spiders were fed flies that had been starved for 48 h in order to empty their guts. Gut emptying of the flies did not reduce the positive effects of the enriched fruit fly media, i.e. the nutritional benefits were not due to nutrients that passed directly through the guts of the flies. Since the nutrients added to the fruit fly media were separated from the spiders that benefited from them by two trophic transformations, this phenomenon was a true tritrophic interaction.