Kim Jensen

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.

Sex-specific effects of protein and carbohydrate intake on reproduction but not lifespan in Drosophila melanogaster

Modest dietary restriction extends lifespan (LS) in a diverse range of taxa and typically has a larger effect in females than males. Traditionally, this has been attributed to a stronger trade‐off between LS and reproduction in females than in males that is mediated by the intake of calories. Recent studies, however, suggest that it is the intake of specific nutrients that extends LS and mediates this trade‐off. Here, we used the geometric framework (GF) to examine the sex‐specific effects of protein (P) and carbohydrate (C) intake on LS and reproduction in Drosophila melanogaster. We found that LS was maximized at a high intake of C and a low intake of P in both sexes, whereas nutrient intake had divergent effects on reproduction. Male offspring production rate and LS were maximized at the same intake of nutrients, whereas female egg production rate was maximized at a high intake of diets with a P:C ratio of 1:2. This resulted in larger differences in nutrient‐dependent optima for LS and reproduction in females than in males, as well as an optimal intake of nutrients for lifetime reproduction that differed between the sexes. Under dietary choice, the sexes followed similar feeding trajectories regulated around a P:C ratio of 1:4. Consequently, neither sex reached their nutritional optimum for lifetime reproduction, suggesting intralocus sexual conflict over nutrient optimization. Our study shows clear sex differences in the nutritional requirements of reproduction in D. melanogaster and joins the growing list of studies challenging the role of caloric restriction in extending LS.

Diffusivity in surficial sediments and benthic mats determined by use of a combined N2O-O2 microsensor

Abstract Diffusional characteristics of two biologically active surface sediments were determined by use of a combined N2O-O2 microsensor. By analyzing changes in the N2O-gradients in these sediments, it was possible to determine the product (φDs) for this species with submillimetre depth resolution, where φ is the porosity and Ds the substrate diffusion coefficient. The (φDs)-value for O2 could be calculated then from (φDs)-value for N2O, because the diffusivity of the two molecules were modified in the same way within the sediment. Both sediments exhibited fine-scale horizontal and vertical variability in diffusion characteristics, and this must be accounted for when analyzing microprofile data. The average (φDs)-value for N2O at 20°C for an estuarine surface sediment was 0.93 × 10−5 cm2 s−1 (at 0–4 mm depth), while the value for the upper 2 mm of a stream sediment covered by a microbial mat was 1.42 × 10−5 cm2 s−1. Biological inactivation and oxidation by exposure to an O2 atmosphere had no effect on the measured (φDs) for the estuarine sediment; however, the value for the sediment covered by a microbial mat, with dense populations of meiofauna, decreased by 20%. The method presented is ideal for measurements of diffusivity at a high spatial resolution in surficial sediments and densely packed microbial communities.

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.

Functional improvement of antibody fragments using a novel phage coat protein III fusion system.

Functional expressions of proteins often depend on the presence of host specific factors. Frequently recombinant expression strategies of proteins in foreign hosts, such as bacteria, have been associated with poor yields or significant loss of functionality. Improvements in the performance of heterologous expression systems will benefit present-day quests in structural and functional genomics where high amounts of active protein are required. One example, which has been the subject of considerable interest, is recombinant antibodies or fragments thereof as expressions of these in bacteria constitute an easy and inexpensive method compared to hybridoma cultures. Such approaches have, however, often suffered from low yields and poor functionality. A general method is described here which enables expressions of functional antibody fragments when fused to the amino-terminal domain(s) of the filamentous phage coat protein III. Furthermore, it will be shown that the observed effect is neither due to improved stability nor increased avidity.

Identification of Keratinocyte-specific Markers Using Phage Display and Mass Spectrometry

Specific molecular markers for various normal and pathogenic cell states and cell types provide knowledge of basic biological systems and have a direct application in targeted therapy. We describe a proteomic method based on the combination of new and improved phage display antibody technologies and mass spectrometry that allows identification of cell type-specific protein markers. The most important features of the method are (i) reduction of experimental noise originating from background binding of phage particles and (ii) isolation of affinity binders after a single round of selection, which assures a high diversity of binders. The method demonstrates, for the first time, the ability to detect, identify, and analyze both secreted and membrane-associated extracellular proteins as well as a variety of different cellular structures including proteins and carbohydrates. The optimized phage display method was applied to analysis of human skin keratinocytes resulting in the isolation of a panel of antibodies. Fourteen of these antibodies were further characterized, half of which predominantly recognized keratinocytes in a screen of a range of different cell types. Three cognate keratinocyte antigens were subsequently identified by mass spectrometry as laminin-5, plectin, and fibronectin. The combination of phage display technology with mass spectrometry methods for protein identification is a general and promising approach for proteomic analysis of cell surface complexity.

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.

Macronutrient balance mediates the growth of sexually selected weapons but not genitalia in male broad‐horned beetles

Summary Condition is defined as the pool of resources available to an individual that can be allocated to fitness-enhancing traits. Consequently, condition could influence developmental trade-offs if any occur. Although many studies have manipulated diet to demonstrate condition-dependent trait expression, few studies have determined the contribution of specific nutrients to condition or trade-offs. We used nutritional geometry to quantify the effects of dietary protein and carbohydrate content on larval performance and the development of adult morphology including body size as well as a primary and secondary sexually selected trait in male broad-horned beetles, Gnatocerus cornutus. We found that offspring survival, development rate and morphological traits were highly affected by dietary carbohydrate content and to a lesser extent by protein content and that all traits were maximized at a protein-to-carbohydrate ratio around 1:2. The absolute size of a secondary sexual character, the mandibles, had a heightened response to the increased availability and ratio of both macronutrients. Male genitalia, in contrast, were relatively insensitive to the increased availability of macronutrients. Overall, while nutrition influenced trait expression, the nutritional requirements of development rate and morphological traits were largely the same and resource acquisition seems to implement only weak trade-offs in this species. This finding contrasts with some resource constraint predictions, as beetles seem able to simultaneously meet the nutritional requirements of most traits.

Identification of phage antibodies toward the Werner protein by selection on Western blots

A procedure was established for selecting phage antibodies (phage‐abs) from phage‐displayed antibody repertoires by panning against proteins, separated by sodium dodecyl phosphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and electroblotted onto nitrocellulose membranes (Western blots). This immobilization strategy is applicable for secondary rounds of panning in selections against semipurified proteins, and directs the selection toward antibodies suitable as immunochemical reagents in Western blots. In model experiments, enrichment factors as high as 1.9 × 105 were obtained in a single round of panning. Furthermore, we demonstrate the application of this approach by selection of phage‐abs recognizing the human Werner protein, which is defective in a premature aging syndrome.

Adaptive contraction of diet breadth affects sexual maturation and specific nutrient consumption in an extreme generalist omnivore

Animals balance their intake of specific nutrients, but little is known about how they do so when foraging in an environment with toxic resources and whether toxic foods promote adaptations that affect life history traits. In German cockroach (Blattella germanica) populations, glucose aversion has evolved in response to glucose‐containing insecticidal baits. We restricted newly eclosed glucose‐averse (GA) and wild‐type (WT) female cockroaches to nutritionally defined diets varying in protein‐to‐carbohydrate (P : C) ratio (3 : 1, 1 : 1, or 1 : 3) or gave them free choice of the 3 : 1 and 1 : 3 diets, with either glucose or fructose as the sole carbohydrate source. We measured consumption of each diet over 6 days and then dissected the females to measure the length of basal oocytes in their ovaries. Our results showed significantly lower consumption by GA compared to WT cockroaches when restricted to glucose‐containing diets, but also lower fructose intake by GA compared to WT cockroaches when restricted to high fructose diets or given choice of fructose‐containing diets. Protein intake was regulated tightly regardless of carbohydrate intake, except by GA cockroaches restricted to glucose‐containing diets. Oocyte growth was completely suppressed in GA females restricted to glucose‐containing diets, but also significantly slower in GA than in WT females restricted to fructose‐containing diets. Our findings suggest that GA cockroaches have adapted to reduced diet breadth through endocrine adjustments which reduce requirements for energetic fuels. Our study illustrates how an evolutionary change in the chemosensory system may affect the evolution of other traits that govern animal life histories.