Proceedings of the Royal Society B: Biological Sciences

Published by Royal Society, The
Online ISSN: 1471-2954
According to the antagonistic pleiotropy theory of ageing, natural selection has favoured genes conferring short-term benefits to the organism at the cost of deterioration in later life. The 'disposable soma' theory expresses this as a life-history strategy in which somatic maintenance is below the level required to prevent ageing, thus enabling higher immediate fertility. It has been argued that a non-ageing strategy will always be bettered by a low but non-zero rate of ageing, because the costs of such ageing will be felt only in the distant future when they are of negligible importance. Here, we examine this argument critically. We find that a non-ageing strategy will be locally optimal if, in the presence of ageing, the onset of deterioration is sufficiently rapid or early. Conversely, ageing will be optimal if deterioration is sufficiently slow or late. As the temporal profile of ageing changes from one of steady deterioration to one involving a sudden loss of vitality after a period of little or no decline, the conditions for a non-ageing strategy to be locally optimal become progressively more stringent. But for all forms of profile considered, conditions can be found for which a strategy involving no ageing is locally optimal.
A map of the southern part of the African continent showing NDVI levels. The study area, South Africa and Lesotho, is delineated with bold black boundaries. NDVI classes are relatively contiguous in the study area, and the areas of intermediate productivity exhibit the greatest surface area and areal coverage. 
Relative species turnover ( sim ) between adjacent grid cells within NDVI classes along the productivity gradient (all species, diamonds; RSQ: first, grey triangles; second, circles; third, squares; fourth, open triangles; regression lines follow the sequence RSQ first, second, third, all species, fourth from top to bottom). 
The relationship between energy availability and species richness (the species-energy relationship) is one of the best documented macroecological phenomena. However, the structure of species distribution along the gradient, the proximate driver of the relationship, is poorly known. Here, using data on the distribution of birds in southern Africa, for which species richness increases linearly with energy availability, we provide an explicit determination of this structure. We show that most species exhibit increasing occupancy towards more productive regions (occurring in more grid cells within a productivity class). However, average reporting rates per species within occupied grid cells, a correlate of local density, do not show a similar increase. The mean range of used energy levels and the mean geographical range size of species in southern Africa decreases along the energy gradient, as most species are present at high productivity levels but only some can extend their ranges towards lower levels. Species turnover among grid cells consequently decreases towards high energy levels. In summary, these patterns support the hypothesis that higher productivity leads to more species by increasing the probability of occurrence of resources that enable the persistence of viable populations, without necessarily affecting local population densities.
Measuring trends in the size of prehistoric populations is fundamental to our understanding of the demography of ancient people and their responses to environmental change. Archaeologists commonly use the temporal distribution of radiocarbon dates to reconstruct population trends, but this can give a false picture of population growth because of the loss of evidence from older sites. We demonstrate a method for quantifying this bias, and we use it to test for population growth through the Holocene of Australia. We used model simulations to show how turnover of site occupation across an archaeological landscape, interacting with erasure of evidence at abandoned sites, can create an increase in apparent site occupation towards the present when occupation density is actually constant. By estimating the probabilities of abandonment and erasure from archaeological data, we then used the model to show that this effect does not account for the observed increase in occupation through the Holocene in Australia. This is best explained by population growth, which was low for the first part of the Holocene but accelerated about 5000 years ago. Our results provide new evidence for the dynamism of non-agricultural populations through the Holocene.
The localization of the human erythrocyte membrane Ins(1,3,4,5)P4 3-phosphatase was investigated by saponin permeabilization of resealed 'isoionic' erythrocyte ghosts. This enzyme is active at the inner face of the plasma membrane, at the same site as a specific 5-phosphatase that degrades both Ins (1,4,5)P3 and Ins(1,3,4,5)P4. In the presence of EDTA, Ins(1,4,5)P3 was the only product of Ins(1,3,4,5)P4 metabolism. However, when Mg2+ was present both the 5-phosphatase and the 3-phosphatase attacked Ins (1,3,4,5)P4, directly forming Ins(1,3,4)P3 and Ins(1,4,5)P3;some Ins(1,4)P2 was also formed as a product of 5-phosphatase attack on the liberated Ins(1,4,5)P3. The Ins(1,3,4,5)P4 3-phosphatase was potently activated by KCl, thus making the route of metabolism of Ins(1,3,4,5)P4 by erythrocyte ghosts strikingly sensitive to variations in ionic strength: at 'cytosolic' K+ and Mg2+ levels, 3-phosphatase activity slightly predominated over 5-phosphatase. Ins(1,3,4,5)P4 3-phosphatase was potently inhibited by Ins-(1,3,4,5,6)P5 and InsP6 at levels lower than those often observed within cells. This leaves open the question as to whether the cellular function of inositol polyphosphate 3-phosphatase is to participate in a physiological cycle that interconverts Ins(1,3,4,5)P4 and Ins(1,4,5)P3 or to metabolize other inositol polyphosphates in the cytosol compartment of cells.
Summary of latencies for InsP3-evokedJfuorescence and conductance changes 
Quantitative, time-resolved measurements have been made of intracellular Ca ion release by inositol 1,4,5-trisphosphate (InsP3) and extracellular ATP in porcine aortic endothelial cells in tissue culture. Intracellular free [Ca] was detected with the calcium dye fluo-3 and InsP3 released intracellularly by photolysis of 'caged' InsP3 in whole-cell voltage-clamped aortic endothelial cells. A rise of [Ca] was recorded at InsP3 concentrations greater than 0.2 microM. The timecourse at low InsP3 concentrations comprised a delay of mean 300 ms (range 266-330 ms), a peak in 2-3 s before declining with a half-time of 5-10 s. The delay and time-to-peak decreased with increasing concentrations of InsP3 over the range 0.2-5 microM. At very high concentrations of InsP3 (> 5 microM), the delay in the Ca response was short, always less than 20 ms. The results are consistent with a direct binding and gating action of InsP3 on the Ca channel of the cellular store. Following InsP3 action there is a refractoriness of the InsP3 Ca release process which recovers with a timecourse of half-time about 30 s. A comparison can be made between the timecourse of InsP3 and extracellular ATP actions. High concentrations of ATP (500 microM) acted with a delay of mean 1.8 s (range 1.2-2.5 s), whereas even moderate concentrations of InsP3 acted much more quickly, suggesting that there are slow coupling steps before or during the production of InsP3 in response to extracellular ATP. Both ATP and InsP3 evoked an increase in membrane conductance to K+, probably via Ca.
Sexual selection through female mate choice exerts a strong selection pressure on males' sexual traits, particularly when direct benefits are involved. In species with male parental care, one would expect sexual selection to favour paternal quality, for instance through selection on morphological structures which promote quality. We experimentally studied the influence of pectoral fins on paternal quality in male three-spined sticklebacks (Gasterosteus aculeatus L.). After reductions of fin area to different degrees, similar-sized males had to perform a complete reproductive cycle in enclosures in the field. The collected data on fanning behaviour and egg development showed that a reduction in pectoral fin size affected paternal quality probably through an increased beat frequency of the pectorals. Thus, pectoral fins can potentially signal paternal quality to choosy females.
We have investigated projectin, a large protein of insect muscles, in Drosophila melanogaster. The 5.3 kilobases of coding sequence reported here contains Class I and Class II motifs characteristic of titin and twitchin, arranged in a three domain ... [II-I-I] [II-I-I] ... pattern. Two mutants mapped to the location of the projectin gene in the 102C subdivision of chromosome 4, lethal(4) 102 CDa and bent-Dominant, have DNA rearrangements within their projectin gene. The lethal(4) 102 CDa mutant has a 141 nucleotide insertion containing stop codons in all three reading frames within an exon sequence, showing that it cannot synthesize normal projectin. Both bent-Dominant and lethal(4) 102 CDa homozygotes die at the completion of embryogenesis because they are unable to escape the egg vitelline membrane. We propose that this hatching failure is due to muscle weakness caused by projectin defects.
Survivorship to pupation of resistant selected and unselected in the parental generation (P R sel, P R ), hybrid (F 1 f, F 1 m) and susceptible (P S ) genotypes feeding on cucumber, 
History of selection of the P R strain.
Least-square mean larval weights ( G s.e.) of resistant selected and unselected in the parental generation (P R sel, P R ), hybrid (F 1 f, F 1 m) and susceptible (P S ) genotypes feeding on cucumber, tomato or pepper. (Significant 
Selection for resistance to insecticides, diseases and parasitoids is assumed to be costly and often requires tradeoffs with reproductive fitness. The costs of resistance, however, are often difficult to measure. Cabbage looper, Trichoplusia ni, a generalist Lepidopteran herbivore, has become highly resistant following the extensive use of the microbial insecticide, Bacillus thuringiensis kurstaki (Bt) in vegetable greenhouses. We compared the growth rate, pupal size and survival of resistant, susceptible and hybrid T. ni larvae fed on tomato, bell pepper and cucumber. Performance was best on cucumber and worst on pepper, and the magnitude of fitness costs associated with Bt resistance increased with declining host plant suitability. This supports the hypothesis that in this system, resistance costs are condition dependent and are greatest in the most stressful environment. Management strategies that rely on the presence of fitness costs to reduce the frequency of resistance genes must consider this variation and should be more successful on crops that are less suitable food plants. In general, condition dependence should be considered in studies designed to measure the costs of resistance.
The ancient mutualism between fungus-growing ants and the fungi they cultivate for food is a textbook example of symbiosis. Fungus-growing ants' ability to cultivate fungi depends on protection of the garden from the aggressive microbes associated with the substrate added to the garden as well as from the specialized virulent garden parasite Escovopsis. We examined ants' ability to remove alien microbes physically by infecting Atta colombica gardens with the generalist pathogen Trichoderma viride and the specialist pathogen Escovopsis. The ants sanitized the garden using two main behaviours: grooming of alien spores from the garden (fungus grooming) and removal of infected garden substrate (weeding). Unlike previously described hygienic behaviours (e.g. licking and self-grooming), fungus-grooming and garden-removal behaviours are specific responses to the presence of fungal pathogens. In the presence of pathogens, they are the primary activities performed by workers, but they are uncommon in uninfected gardens. In fact, workers rapidly eliminate Trichoderma from their gardens by fungus grooming and weeding, suggesting that these behaviours are the primary method of garden defence against generalist pathogens. The same sanitary behaviours were performed in response to the presence of the specialist pathogen Escovopsis. However, the intensity and duration of these behaviours were much greater in this treatment. Despite the increased effort, the ants were unable to eliminate Escovopsis from their gardens, suggesting that this specialized pathogen has evolved counter-adaptations in order to overcome the sanitary defences of the ants.
E¡ects of oxytetracycline treatment of cattle on O. ochengi worm number and motility control oxytetracycline 
Light microscopy of uterine embryonic stages (b, upper left and upper right insets) and histological sections (a, b bottom image, c, d ) of tetracycline-treated worms. (a) Enlarged hypodermis (hyp) and formation of vacuoles in oogonia (oog), at two mpt ( £ 600). (b) Vacuolated (upper left inset) and degenerating horseshoe-shaped and coiled micro¢lariae (upper right inset) at two mpt (phase contrast); below, degenerating intrauterine micro¢lariae (di¡erential interference contrast) ( £ 630). (c) Invasion of eosinophils (eos) near degenerating female at three mpt. Inner worm is top left with thin hypodermis containing a degenerating nucleus (n) and below it cuticular debris is being shed into the hypercellular nodule tissue ( £ 440). (d ) Collapsed, dying or dead female with empty uteri at six mpt (di¡erential interference contrast) ( £ 150).  
Filarial nematodes are important and widespread parasites of animals and humans. We have been using the African bovine parasite Onchocerca ochengi as a chemotherapeutic model for O. volvulus, the causal organism of 'river blindness' in humans, for which there is no safe and effective drug lethal to adult worms. Here we report that the antibiotic, oxytetracycline is macrofilaricidal against O. ochengi. In a controlled trial in Cameroon, all adult worms (as well as microfilariae) were killed, and O. ochengi intradermal nodules resolved, by nine months' post-treatment in cattle treated intermittently for six months. Adult worms removed from concurrent controls remained fully viable and reproductively active. By serial electron-microscopic examination, the macrofilaricidal effects were related to the elimination of intracellular micro-organisms, initially abundant. Analysis of a fragment of the 16S rRNA gene from the O. ochengi micro-organisms confirmed them to be Wolbachia organisms of the order Rickettsiales, and showed that the sequence differed in only one nucleotide in 858 from the homologous sequence of the Wolbachia organisms of O. volvulus. These data are, to our knowledge, the first to show that antibiotic therapy can be lethal to adult filariae. They suggest that tetracycline therapy is likely to be macrofilaricidal against O. volvulus infections in humans and, since similar Wolbachia organisms occur in a number of other filarial nematodes, against those infections too. In that the elimination of Wolbachia preceded the resolution of the filarial infections, they suggest that in O. ochengi at least, the Wolbachia organisms play an essential role in the biology and metabolism of the filarial worm.
During an infection, malaria parasites compete for limited amounts of food and enemy-free space. Competition affects parasite growth rate, transmission and virulence, and is thus important for parasite evolution. Much evolutionary theory assumes that virulent clones outgrow avirulent ones, favouring the evolution of higher virulence. We infected laboratory mice with a mixture of two Plasmodium chabaudi clones: one virulent, the other avirulent. Using real-time quantitative PCR to track the two parasite clones over the course of the infection, we found that the virulent clone overgrew the avirulent clone. However, host genotype had a major effect on the outcome of competition. In a relatively resistant mouse genotype (C57B1/6J), the avirulent clone was suppressed below detectable levels after 10 days, and apparently lost from the infection. By contrast, in more susceptible mice (CBA/Ca), the avirulent clone was initially suppressed, but it persisted, and during the chronic phase of infection it did better than it did in single infections. Thus, the qualitative outcome of competition depended on host genotype. We suggest that these differences may be explained by different immune responses in the two mouse strains. Host genotype and resistance could therefore play a key role in the outcome of within-host competition between parasite clones and in the evolution of parasite virulence.
When local populations are genetically differentiated from one another and partially inbred, as typically occurs in subdivided populations, immigrant genomes are predicted to be at a frequency-dependent fitness advantage due to heterosis (hybrid vigour) in their descendants. We tested this prediction with pedigreed laboratory populations of the butterfly Bicyclus anynana and report here on a rapid increase over five generations in the contribution of an initially rare immigrant genome to the local population gene pool. The replicated experimental design, including immigrant controls, demonstrates that the mechanism underlying immigrant genome spread is heterosis, and that the advantage to the immigrant genes is sustained over several generations. Our result suggests that effective migration rates may often be much higher than the numbers of individual migrants assumed by classical population genetics models, with implications for the persistence and evolution of metapopulations.
Estimated S. avenae population structure. Each aphid is represented by a thin vertical line, which is partitioned into K segments, representing the individual's estimated membership fractions in K clusters. Black lines separate individuals of different populations labelled with the host plant. 1, Yorkshire fog; 2, cocksfoot; 3, bent; 4, tall wild oat; 5, bulbous oatgrass; 6, oat; 7, barley; 8, triticale; 9, wheat. Tribe of host plants (Poeae, Aveneae, Triticeae) is specified under the figure. 
Mean proportion of ancestry calculated for each host population, from results obtained by clustering analysis on individuals for KZ9 clusters (named C1-C9). 
Habitats in agroecosystems are ephemeral, and are characterized by frequent disturbances forcing pest species to successively colonize various hosts belonging either to the cultivated or to the uncultivated part of the agricultural landscape. The role of wild habitats as reservoirs or refuges for the aphid Sitobion avenae that colonize cultivated fields was assessed by investigating the genetic structure of populations collected on both cereal crops (wheat, barley and oat) and uncultivated hosts (Yorkshire fog, cocksfoot, bulbous oatgrass and tall oatgrass) in western France. Classical genetic analyses and Bayesian clustering algorithms indicate that genetic differentiation is high between populations collected on uncultivated hosts and on crops, revealing a relatively limited gene flow between the uncultivated margins and the cultivated part of the agroecosystem. A closer genetic relatedness was observed between populations living on plants belonging to the same tribe (Triticeae, Poeae and Aveneae tribes) where aphid genotypes appeared not to be specialized on a single host, but rather using a group of related plant species. Causes of this ecological differentiation and its implications for integrated pest management of S. avenae as cereals pest are discussed.
Hox gene expression pattern and the evolution of the female reproductive tract. (a) HoxA-13 to HoxA-9 are located at the 5 0 end of the HoxA cluster and are expressed in the same regions in the adult as in the embryo: HoxA-13 (green), HoxA-11 (yellow), HoxA-10 (orange) and HoxA-9 (blue). (b) Tetrapod phylogeny showing representative female reproductive systems from each group (amphibian ovaries shown only on the left). 
Phylogeny of the species in this study. The maximum-likelihood estimate of the non-synonymous-to-synonymous substitution rate ratio is given above the branch (x ¼ d N /d S ) for HoxA-11 (a) and HoxA-13 (b). The three-ratios model was used to estimate d N and d S for both genes. The stem lineages of therians (T) and eutherians (E) are labelled. The estimate of the background ratio (x 0 ) is shown above the root of each tree. Branches inferred to be under positive selection are shown in bold and are drawn in proportion to the estimate of their length, defined as the number of subsitutions per codon. Ã An estimate of x for this branch could not be made because d S ¼ 0; however, 5.2 non-synonymous changes occurred along this branch. 
The evolution of morphological characters is mediated by the evolution of developmental genes. Evolutionary changes can either affect cis-regulatory elements, leading to differences in their temporal and spatial regulation, or affect the coding region. Although there is ample evidence for the importance of cis-regulatory evolution, it has only recently been shown that transcription factors do not remain functionally equivalent during evolution. These results suggest that the evolution of transcription factors may play an active role in the evolution of development. To test this idea we investigated the molecular evolution of two genes essential for the development and function of the mammalian female reproductive organs, HoxA-11 and HoxA-13. We predicted that if coding-region evolution plays an active role in developmental evolution, then these genes should have experienced adaptive evolution at the origin of the mammalian female reproductive system. We report the sequences of HoxA-11 from basal mammalian and amniote taxa and analyse HoxA-11 and HoxA-13 for signatures of adaptive molecular evolution. The data demonstrate that these genes were under strong positive (directional) selection in the stem lineage of therian and eutherian mammals, coincident with the evolution of the uterus and vagina. These results support the idea that adaptive evolution of transcription factors can be an integral part in the evolution of novel structures.
Human melanopsin in HEK293 cells. (a) Western blot of opsin protein expressed and extracted from HEK293 cells. Blots were stripped and re-probed with an antibody against total ERK protein to indicate protein amount. Predicted sizes of the opsin proteins as calculated with are provided at foot of blot. (b) Immunocytochemistry photomicrograph showing detection of human melanopsin expressed in HEK293 cells and labelled with a 1D4 antibody (green). Cells are also stained with DAPI, a blue nuclear stain. Melanopsin can be seen mostly localized at the cell membrane and in cytoplasmic inclusions. Scale bar = 6 µm. (c) A luminescent biosensor was used to monitor changes in cAMP production. The camera flash (yellow arrow) induced a large increase in cAMP-dependent luminescence indicative of Gs activity in cells expressing JellyOp but not those expressing human Opn4. (d) In this assay, there was no difference in the change in luminescence following light exposure in cells expressing human Opn4 and no opsin controls (t-test p > 0.05; n ≥ 3). (e) Reductions in cAMP as a result of Gi/o activity are measured by artificially raising cAMP with forskolin (black arrow) before exposing cells to light (yellow arrow). Human rod opsin (Rh1) induces a prolonged reduction in cAMP production. Luminescence was normalized to values immediately following light exposure (n ≥ 6). (f) Comparison of the reduction of forskolin-induced cAMP production 1 min following light in human melanopsin or Rh1 expressing cells treated or untreated with 100 ng ml−1 pertussis toxin (ptx). A pairwise comparison shows the light-dependent reduction is significantly reduced when treated with ptx, a Gi/o-specific inhibitor (paired t-test; **p < 0.01 both opsins). The inhibition appears greater in Rh1 expressing cells. (g) Aequorin luminescence in HEK293 cells expressing opsins as labelled. A camera flash of white light was applied (yellow arrow) and aequorin responses in a 96 well plate reader were monitored. n ≥ 4 except human rod opsin (Rh1), n = 2. (h) There was a significant difference in peak of luminescence immediately following light exposure between no opsin controls and human melanopsin (Opn4; t-test ***p < 0.001). All plots show mean ± s.e.m.
G-protein coupling of melanopsins from other species. Gq/11 assays showed robust calcium-dependent increases in aequorin luminescence from mouse and amphioxus Opn4 (a) and chicken Opn4m and Opn4x (b). (c) Amphioxus and mouse Opn4 showed a small inhibition of forskolin (black arrow)-induced luminescence following light exposure (grey arrow) indicative of a minor Gi/o response. (d) Conversely, both chicken Opn4x and Opn4m cause an increase in cAMP reporter luminescence following forskolin and light exposure. (e) This light-dependent increase in cAMP production (indicative of Gs coupling) is also measurable when no forskolin is first added. All plots show mean ± s.e.m.; n = ≥ 3. (f) The maximum difference in cAMP reporter luminescence before and after light exposure is significantly greater in Opn4m but not Opn4x expressing cells compared with when no opsin is expressed (Mann Whitney test *p < 0.05). Mean ± s.e.m., n = 4.
Spectral sensitivity of melanopsin. Action spectra for (a) mouse and (b) human melanopsins were defined by plotting the relative sensitivity at each wavelength (mean ± s.e.m. of the three replicate irradiance response curves). The action spectra were fit with the predicted absorbance spectrum of opsin : vitamin A-based photopigments with λmax between 450 and 550 nm, and the best fit assessed by minimizing sums of squares. Mouse melanopsin best matches a template with λmax 484 nm, and human melanospin λmax 479 nm. Sum of squares for comparisons between our data and opsin templates with λmax between 470 and 490 nm are shown as an indication of the confidence of these estimates for (c) mouse and (d) human melanopsin.
A subset of mammalian retinal ganglion cells expresses an opsin photopigment (melanopsin, Opn4) and is intrinsically photosensitive. The human retina contains melanopsin, but the literature lacks a direct investigation of its spectral sensitivity or G-protein selectivity. Here, we address this deficit by studying physiological responses driven by human melanopsin under heterologous expression in HEK293 cells. Luminescent reporters for common second messenger systems revealed that light induces a high amplitude increase in intracellular calcium and a modest reduction in cAMP in cells expressing human melanopsin, implying that this pigment is able to drive responses via both Gq and Gi/o class G-proteins. Melanopsins from mouse and amphioxus had a similar profile of G-protein coupling in HEK293 cells, but chicken Opn4m and Opn4x pigments exhibited some Gs activity in addition to a strong Gq/11 response. An action spectrum for the calcium response in cells expressing human melanopsin had the predicted form for an opsin : vitamin A1 pigment and peaked at 479 nm. The G-protein selectivity and spectral sensitivity of human melanopsin is similar to that previously described for rodents, supporting the utility of such laboratory animals for developing methods of manipulating this system using light or pharmacological agents.
Size comparison of lower molar teeth of Early Cretaceous tribosphenic mammals from the Antlers Formation and correlative Trinity Group units, TX. Abbreviations: C1, Comanchea (SMU 73070, right m x ); C2, Comanchea (SMU 73206, left m x ); H1, Holoclemensia (SMU 61727, left m x ); H2, Holoclemensia (SMU 62131, left m x ); H3, Holoclemensia (SMU 61726, left m 4 ); K, Kermackia (SMU 62398, right m x ); P1, Pappotherium (PM 948, m 3 or m 4 (?)); P2, Pappotherium (PM 965, m 2 (?)); S1, Slaughteria (SMU 61992, left t 3 ); S2, Slaughteria (SMU 61992, left t 4 ); T, Trinititherium (SMU 61728, left m x ). PM, Field Museum of Natural History, Chicago. Measurements of PM 948 and PM 965 are from Turnbull (1971). 
Living placental and marsupial mammals (therians) use distinctive tooth-replacement patterns that have not yet been traced back fully to their time of divergence in the Early Cretaceous (>100 Myr ago). Slaughteria eruptens, a small 110 Myr old fossil mammal from Texas, USA, is near the base of that divergence. Using ultra-high-resolution X-ray CT analysis we demonstrate that Slaughteria preserves an unrecognized pattern of tooth replacement with simple posterior premolars replacing molariform precursors. Differing from both placentals that have a more complex posterior adult premolar, and from marsupials, in which only one premolar is replaced, Slaughteria provides the first direct evidence of a tooth-replacement pattern that is plausible for the common ancestor of all therians. By our interpretation Slaughteria has only one adult molar in place and contains two mental foramina in the jaw, thus changing characters that are critical to reconstruction of mammalian relationships and to species discrimination and interpretations of diversity for Early Cretaceous mammals.
Inherited bacterial symbionts from the genus Wolbachia have attracted much attention by virtue of their ability to manipulate the reproduction of their arthropod hosts. The potential importance of these bacteria has been underlined by surveys, which have estimated that 17% of insect species are infected. We examined whether these surveys have systematically underestimated the proportion of infected species through failing to detect the low-prevalence infections that are expected when Wolbachia distorts the sex ratio. We estimated the proportion of species infected with Wolbachia within Acraea butterflies by testing large collections of each species for infection. Seven out of 24 species of Acraea were infected with Wolbachia. Four of these were infected with Wolbachia at high prevalence, a figure compatible with previous broad-scale surveys, whilst three carried low-prevalence infections that would have had a very low likelihood of being detected by previous sampling methods. This led us to conclude that sex-ratio-distorting Wolbachia may be common in insects that have an ecology and/or genetics that permit the invasion of these parasites and that previous surveys may have seriously underestimated the proportion of species that are infected.
Context-dependent life-history variation. Factor plots for a factor analysis of six life-history traits for soil mites living in environments characterized by high and low food availability. The angle between vectors reflects the correlations between the traits (less than 908, positive correlations; greater than 908, negative correlations; 908, no correlation). Under low food conditions, the size of the egg an individual hatches from (egg length) is strongly positively correlated with survival to adulthood (recruitment) and age at maturity, negatively with size at maturity but uncorrelated with adult survival (survival) or fecundity. Conversely, under high food conditions, egg size is strongly correlated with fecundity and adult survival and not with juvenile survival to recruitment or size and age at maturity. Figure after fig. 4 in Plaistow et al. (2006).  
Population dynamics result from the interplay of density-independent and density-dependent processes. Understanding this interplay is important, especially for being able to predict near-term population trajectories for management. In recent years, the study of model systems-experimental, observational and theoretical-has shed considerable light on the way that the both density-dependent and -independent aspects of the environment affect population dynamics via impacting on the organism's life history and therefore demography. These model-based approaches suggest that (i) individuals in different states differ in their demographic performance, (ii) these differences generate structure that can fluctuate independently of current total population size and so can influence the dynamics in important ways, (iii) individuals are strongly affected by both current and past environments, even when the past environments may be in previous generations and (iv) dynamics are typically complex and transient due to environmental noise perturbing complex population structures. For understanding population dynamics of any given system, we suggest that 'the devil is in the detail'. Experimental dissection of empirical systems is providing important insights into the details of the drivers of demographic responses and therefore dynamics and should also stimulate theory that incorporates relevant biological mechanism.
The strategies proposed for delaying the development of resistance to the Bacillus thuringiensis toxins produced by transgenic maize require high levels of gene flow between individuals feeding on transgenic and refuge plants. The European corn borer Ostrinia nubilalis (Hübner) may be found on several host plants, which may act as natural refuges. The genetic variability of samples collected on sagebrush (Artemisia sp.), hop (Humulus lupulus L.) and maize (Zea mays L.) was studied by comparing the allozyme frequencies for six polymorphic loci. We found a high level of gene flow within and between samples collected on the same host plant. The level of gene flow between the sagebrush and hop insect samples appeared to be sufficiently high for these populations to be considered a single genetic panmictic unit. Conversely, the samples collected on maize were genetically different from those collected on sagebrush and hop. Three of the six loci considered displayed greater between-host-plant than within-host-plant differentiation in comparisons of the group of samples collected on sagebrush or hop with the group of samples collected on maize. This indicates that either there is genetic isolation of the insects feeding on maize or that there is host-plant divergent selection at these three loci or at linked loci. These results have important implications for the potential sustainability of transgenic insecticidal maize.
Female colour forms and broods for mapping used in this study. (a) Wing patterns and their nomenclature for female forms used in crosses. (b) The female-informative Brood 99 involving the segregation of forms cenea (H c ) and hippocoon (h), with the genetic interpretation of parents and offspring and the number of offspring. (c) The maleinformative Brood 59.  
Segregation pattern and genetic distances in the H region. (a) Co-segregation of various alleles of the AFLP markers and inv in two types of female offspring (cenea above, hippocoon below) of the male-informative Brood 59. The number of female offspring (n) maintaining the parental phase or showing recombination is also shown, with inferred crossover events marked in the figure. (b) A map of the H genome region based on co-segregation data from 133 offspring of four female-informative families.  
The swallowtail butterfly, Papilio dardanus, is an iconic example of a polymorphic Batesian mimic. The expression of various female-limited colour forms is thought to be controlled by a single autosomal locus, termed H, whose function in determining the wing pattern remains elusive. As a step towards the physical mapping of H, we established a set of 272 polymorphic amplified fragment length polymorphism (AFLP) markers (EcoRI-MseI). Segregation patterns in a 'female-informative' brood (exploiting the absence of crossing over in female Lepidoptera) mapped these AFLPs to 30 linkage groups (putative chromosomes). The difference between the hippocoon and cenea female forms segregating in this family resides on a single one of these linkage groups, defined by 14 AFLPs. In a 'male-informative' cross (markers segregating within a linkage group), a pair of AFLPs co-segregated closely with the two female forms, except in four recombinants out of 19 female offspring. Linkage with these AFLP markers using four further female-informative families demonstrated that the genetic factor determining other morphs (poultoni, lamborni and trimeni) also maps to this same linkage group. The candidate gene invected, obtained in a screen for co-segregation of developmental genes with the colour forms, resides in a 13.9 cM interval flanked by the two AFLP markers. In the male-informative family invected co-segregated perfectly with the hippocoon/cenea factor, despite the four crossovers with the AFLPs. These findings make invected, and possibly its closely linked paralogue engrailed, strong candidates for H. This is supported by their known role in eyespot specification in nymphalid butterfly wings.
Ebert (1994) has proposed the rule that parasites are, with few exceptions, more infective to sympatric hosts than to allopatric hosts. We test this rule using field data for schistosome infections of planorbid snails and find that, although sympatric parasite-host combinations do tend to be more compatible, there are exceptions where particular allopatric parasite-host populations are significantly more compatible. We develop a mathematical model of the dynamics of the parasite-host interaction where parasite infectivity and host susceptibility are defined by the matching of genotypes in a diploid system, The model predicts dynamic polymorphisms where parasite allele frequencies track host allele frequencies but with a lag. Because of this lag, it is possible for allopatric combinations to be more compatible than sympatric combinations. Any 'rule' that precludes this possibility is unlikely to prove robust.
Long-term LBM and temperature reconstructions for the European Alps. (a) MXD-based LBM outbreak reconstruction since AD 832. Time-series is the age-corrected difference series between gap-filled and original MXD data (for details on gap-filling and age-correction procedures, see the electronic supplementary material). Values less than K0.005 g cm K3 are shown. (b) The temperature model (white curve) is shown together with the standard error (coloured band) derived from the fit with instrumental data. Dashed lines indicate the last LBM mass outbreak in 1981 (vertical) and the upper standard error limit recorded in the late ninth century (horizontal). 
Temporal characteristics of the long-term LBM reconstruction shown in figure 3a. (a) MTM spectrum (Mann & Lees 1996) of the reconstruction. Grey curves are estimated significance levels. (b) Estimated autocorrelation functions (acf ) for lags 0-20 of the reconstruction. Colours indicate different periods (full period and segments) used for acf calculation. Dashed curve is the 95% significance level. 
Characteristics of LBM outbreak events reconstructed over the past millennium (see table 1 for complete list). (a) Return time record calculated from the 123 LBM defoliation events. Colours indicate a classification of the intensity of outbreaks, i.e. the reduction of wood density (in g cm K3 ) caused by LBM. Dashed curve in the late twentieth century refers to the recent absence in mass outbreaks since 1981. Horizontal dashed curves are the two standard deviations. (b) Arithmetic mean and standard error (bars), and coefficient of variation computed for the full return time record (AD 832-2004) and six subperiods. Dashed curves in recent centuries result from including a 25-year return time interval since 1981, i.e. an LBM mass outbreak in 2006 was assumed. (c) Relative frequency distribution of the return time data. Colours indicate different periods. Note that a single event transforms to bars of different relative heights (e.g. all spikes in classes above 14 years correspond to single events). 
The long-term history of Zeiraphera diniana Gn. (the larch budmoth, LBM) outbreaks was reconstructed from tree rings of host subalpine larch in the European Alps. This record was derived from 47513 maximum latewood density measurements, and highlights the impact of contemporary climate change on ecological disturbance regimes. With over 1000 generations represented, this is the longest annually resolved record of herbivore population dynamics, and our analysis demonstrates that remarkably regular LBM fluctuations persisted over the past 1173 years with population peaks averaging every 9.3 years. These regular abundance oscillations recurred until 1981, with the absence of peak events during recent decades. Comparison with an annually resolved, millennium-long temperature reconstruction representative for the European Alps (r=0.72, correlation with instrumental data) demonstrates that regular insect population cycles continued despite major climatic changes related to warming during medieval times and cooling during the Little Ice Age. The late twentieth century absence of LBM outbreaks, however, corresponds to a period of regional warmth that is exceptional with respect to the last 1000+ years, suggesting vulnerability of an otherwise stable ecological system in a warming environment.
Relationship between mean nesting trait values in each nesting season of the painted turtle, C. picta, in Illinois, USA, and the HDD from September to April preceding the nesting season (Schwanz & Janzen 2008). (a) average percentage south þ west vegetation cover of first nest laid by females in different years in response to HDD ( y ¼ 20.0075x þ 134.52, R 2 ¼ 0.289, p , 0.07). (b) Average Julian nest date of the first nest laid by females in different years in response to HDD ( y ¼ 0.0063x þ 124.94, R 2 ¼ 0.364, p , 0.02). 
Nesting behaviour is critical for reproductive success in oviparous organisms with no parental care. In organisms where sex is determined by incubation temperature, nesting behaviour may be a prime target of selection in response to unbalanced sex ratios. To produce an evolutionary change in response to sex-ratio selection, components of nesting behaviour must be heritable. We estimated the field heritability of two key components of nesting behaviour in a population of painted turtles (Chrysemys picta) with temperature-dependent sex determination by applying the 'animal model' to a pedigree reconstructed from genotype data. We obtained estimates of low to non-detectable heritability using repeated records across all environments. We then determined environment-specific heritability by grouping records with similar temperatures for the winter preceding the nesting season, a variable known to be highly associated with our two traits of interest, nest vegetation cover and Julian date of nesting. The heritability estimates of nest vegetation cover and Julian date of nesting were qualitatively highest and significant, or nearly so, after hot winters. Additive genetic variance for these traits was not detectable after cold winters. Our analysis suggests that the potential for evolutionary change of nesting behaviour may be dependent on the thermal conditions of the preceding winter, a season that is predicted to be especially subject to climate change.
Number of times (out of 1000) the sexuals won as function of the parameters (a) d, (b) L, (c) n and (d ) G. In simulations where these parameters are not varied, their values were chosen to be LZ10 (only for varying n, where larger L require too long simulation times), dZ5, GZ10, and nZ10. 
Variance in trait value of the winning mode of reproduction for the simulations shown in the first graph of figure 1. plus, asexuals; cross, sexuals. 
We present a model for the maintenance of sexual reproduction based on the availability of resources, which is the strongest factor determining the growth of populations. The model compares completely asexual species to species that switch between asexual and sexual reproduction (sexual species). Key features of the model are that sexual reproduction sets in when resources become scarce, and that at a given place only a few genotypes can be present at the same time. We show that under a wide range of conditions the sexual species outcompete the asexual ones. The asexual species win only when survival conditions are harsh and death rates are high, or when resources are so little structured or consumer genotypes are so manifold that all resources are exploited to the same extent. These conditions largely represent the conditions in which sexuals predominate over asexuals in the field.
Dorsal wing surface of representative males of each group of manipulated male Bicyclus anynana butter ies. (a) Males that were damaged away from the focal area on both left and right side (group 1); (b) focal damage on one side (left or right) and non-focal on the other (group 2); and (c) focal damage on both sides (group 3). The ventral wing surface has no eyespots. The two dorsal eyespots, A and P, and the interfocal distance as an index of wing size (WS) are indicated.
Cumulative relative frequency (´ 100%) distributions of (a) total eyespot size (i.e. size A 1 size P), (b) interfocal distance (WS), (c) asymmetry values (R–L) of A eyespots, and (d ) asymmetry values (R–L) of P eyespots. Solid lines refer to the total dataset of males (n = 228), dashed lines refer to control males in which eyespot size was not manipulated (n = 76).  
The eyespots on the ventral wings of Bicyclus anynana butterflies are exposed when at rest and interact with predators. Those on the dorsal surface are not exposed in this way, and may be involved in courtship and mate choice. In this study, we examined whether the size and fluctuating asymmetry (FA) of dorsal eyespots are reliable signals of male quality. High developmental stability is considered to result in low FA, and to be associated with high quality. Individuals of high quality are predicted to produce sexually selected traits that are large and symmetrical, at a relatively low cost. In this study, we manipulated eyespot development to uncouple eyespot size and FA in order to examine their independent roles in signalling to the female. Individual females in cages were given the choice between two or three males differing in eyespot traits. The results indicate that although size per se of the eyespots is used as a signal, FA and wing size are not. We discuss the use of FA in studies of sexual selection and aspects of sexual selection on dorsal eyespot size.
Results of homogeneity tests for the 45 replicated experiments in the meta-analysis. (Effect size is calculated as 
Mean effect sizes of prey in replicated predator manipulation experiments. (a) Effects of native and introduced predators and (b) effects of native and introduced predators, with the effects of introduced predators divided between Australian experiments and experiments conducted elsewhere. Effect size is calculated as Hedges' d. Bars represent 95% bias-corrected confidence intervals. 
Mean effect sizes of prey in unreplicated predator manipulation experiments. (a) Effects of native and introduced predators and (b) effects of native and introduced predators, with the effects of introduced predators divided between Australian experiments and experiments conducted elsewhere. Effect size presented as back-transformed ln(X e / X c ), where X e and X c are the treatment and control prey responses, respectively. Bars represent 95% confidence intervals. 
Alien predators are widely considered to be more harmful to prey populations than native predators. To evaluate this expectation, we conducted a meta-analysis of the responses of vertebrate prey in 45 replicated and 35 unreplicated field experiments in which the population densities of mammalian and avian predators had been manipulated. Our results showed that predator origin (native versus alien) had a highly significant effect on prey responses, with alien predators having an impact double that of native predators. Also the interaction between location (mainland versus island) and predator origin was significant, revealing the strongest effects with alien predators in mainland areas. Although both these results were mainly influenced by the huge impact of alien predators on the Australian mainland compared with their impact elsewhere, the results demonstrate that introduced predators can impose more intense suppression on remnant populations of native species and hold them further from their predator-free densities than do native predators preying upon coexisting prey.
(a) Observed tri-locus hybrid index distribution in backcrosses (pooling the two backcrosses, BC1 and BC2, over the two sampling times, 36 h and D200), expected hybrid index distribution from single-locus frequencies with the assumption of no linkage and expected hybrid index distribution from single-locus Mendelian expectations and no linkage. (b) The deviations of the observed distribution from the Mendelian expectation with no linkage and the expectation from single-locus frequencies and no linkage. 
(a) Tri-locus hybrid index distributions in the F 2 generation, 36 h and 200 days after fertilization; and (b) deviation from the Mendelian expectation with no linkage in the D200 sample. 
We studied the genetic basis of post-zygotic isolation in the marine mussels Mytilus edulis and Mytilus galloprovincialis. Evidence was obtained for a high number of recessive Dobzhansky-Muller substitutions in the genome of these two mussel taxa. We analysed the segregation of unlinked diagnostic markers in the progeny of two backcrosses and an F2 cross, 36 h and 200 days after fertilization. Directional selection favouring M. galloprovincialis genotypes was observed in both kinds of cross. In the F2, epistatic interactions between each pair of chromosome fragments mapped by the markers were identified in addition. Our results imply that homozygous-homozygous interactions are required for breakdown of coadaptation, in accordance with the dominance theory of post-zygotic isolation. Endogenous post-zygotic selection distributed over many loci throughout the genome provides the missing factor explaining the astonishing persistence and strength of barriers to neutral introgression in such a dispersive taxon as Mytilus.
Frequency distributions of the corrected assignment index (AI c ) for 64 females (dark bars above axis) and 76 males (light bars below axis) sampled from 15 populations near Lausanne. AI c values differed among sexes, females having on average negative values, and males positive values. This implies that immigration was female biased in the populations studied. This bias is also reflected in the larger variance of the female distribution, and in its possible bimodality which, suggests the existence of two distinct classes (immigrants vs locally born individuals). 
We investigated dispersal patterns in the monogamous Crocidura russula, based both on direct field observations (mark-recapture data) and on genetic analyses (microsatellite loci). Natal dispersal was found to be low. Most juveniles settled within their natal territory or one immediately adjacent. Migration rate was estimated to two individuals per year and per population. The correlation between genetic and geographical distances over a 16 km transect implies that migration occurs over short ranges. Natal dispersal was restricted to first-litter juveniles weaned in early May; this result suggests a direct dependence of dispersal on reproductive opportunities. Natal dispersal was highly female biased, a pattern unusual among mammals. Its association with monogamy provides support for the resource-competition model of dispersal. Our results demonstrate that a state-biased dispersal can be directly inferred from microsatellite genotype distributions, which opens new perspectives for empirical studies in this area.
Comparison of the numbers of species of Neotemperate, Neotropical and Palaeotemperate insects and arachnids testing positive for Wolbachia 
Wolbachia are a group of cytoplasmically inherited bacteria that cause reproduction alterations in arthropods, including parthenogenesis, reproductive incompatibility, feminization of genetic males and male killing. Previous general surveys of insects in Panama and Britain found Wolbachia to be common, occurring in 16-22% of species. Here, using similar polymerase chain reaction methods, we report that 19.3% of a sample of temperate North American insects are infected with Wolbachia, a frequency strikingly similar to frequencies found in two other studies in widely separated locales. The results may indicate a widespread equilibrium of Wolbachia infection frequencies in insects whose maintenance remains to be explained. Alternatively, Wolbachia may be increasing in global insect communities. Within each of the three geographic regions surveyed, Hymenoptera are more frequently infected with A group Wolbachia and Lepidoptera more frequently infected with B group Wolbachia.
According to the theory of mate choice based on heterozygosity, mates should choose each other in order to increase the heterozygosity of their offspring. In this study, we tested the 'good genes as heterozygosity' hypothesis of mate choice by documenting the mating patterns of wild Atlantic salmon (Salmo salar) using both major histocompatibility complex (MHC) and microsatellite loci. Specifically, we tested the null hypotheses that mate choice in Atlantic salmon is not dependent on the relatedness between potential partners or on the MHC similarity between mates. Three parameters were assessed: (i) the number of shared alleles between partners (x and y) at the MHC (M(xy)), (ii) the MHC amino-acid genotypic distance between mates' genotypes (AA(xy)), and (iii) genetic relatedness between mates (r(xy)). We found that Atlantic salmon choose their mates in order to increase the heterozygosity of their offspring at the MHC and, more specifically, at the peptide-binding region, presumably in order to provide them with better defence against parasites and pathogens. This was supported by a significant difference between the observed and expected AA(xy) (p = 0.0486). Furthermore, mate choice was not a mechanism of overall inbreeding avoidance as genetic relatedness supported a random mating scheme (p = 0.445). This study provides the first evidence that MHC genes influence mate choice in fish.
Females often prefer males with elaborate traits, even when they receive no direct benefits from their choice. In such situations, mate discrimination presumably has genetic advantages; selective females will produce offspring of higher genetic quality. Over time, persistent female preferences for elaborate secondary-sexual traits in males should erode genetic variance in these traits, eventually eliminating any benefit to the preferences. Yet, strong female preferences persist in many taxa. This puzzle is called the lek paradox and raises two primary questions: do females obtain genetic benefits for offspring by selecting males with elaborate secondary-sexual characteristics and, if so, how is the genetic variation in these male traits maintained? We suggest that indirect genetic effects may help to resolve the lek paradox. Maternal phenotypes, such as habitat selection behaviours and offspring provisioning, often influence the condition and the expression of secondary-sexual traits in sons. These maternal influences are commonly genetic based (i.e. they are indirect genetic effects). Females choosing mates with elaborate traits may receive 'good genes' for daughters in the form of effective maternal characteristics. Recognizing the significance of indirect genetic effects may be important to our understanding of the process and consequences of sexual selection.
Map showing the two sampling regions in Denmark, Jutland and Lolland. The populations units defined by coherent groups of ponds are indicated with larger grey circles, whereas the black circles indicate possible connections between ponds or populations but the actual connections are unknown.
A genetic study of the European tree frog, Hyla arborea, in Denmark was undertaken to examine the population structure on mainland Jutland and the island of Lolland after a period of reduction in suitable habitat and population sizes. The two regions have experienced the same rate of habitat loss but fragmentation has been more severe on Lolland. Genetic variation based on 12 polymorphic DNA microsatellites was analysed in 494 tree frogs sampled from two ponds in Jutland and 10 ponds on Lolland. A significant overall deviation from Hardy-Weinberg expectations could be attributed to three ponds, all on Lolland. This was most probably caused by an inbreeding effect reducing fitness, which was supported by the observed significant negative correlation between larva survival and mean F(IS) value and mean individual inbreeding coefficient. A significant reduction in genetic variation (bottleneck) was detected in most of the ponds on Lolland. Population-structure analysis suggested the existence of at least 11 genetically different populations, corresponding to most of the sampled population units. The results indicated that the populations were unique genetic units and could be used to illustrate the migration pattern between newly established ponds arisen either by natural colonization of tree frogs or by artificial introduction. A high degree of pond fidelity in the tree frogs was suggested. A severe fragmentation process reducing population size and fitness within some of the populations probably caused the significant reduction in genetic variation of tree frog populations on Lolland.
Adult ivory gull feeding on a seal carcass, Resolute Bay, Nunavut, Canada, 10 June 1989. Photograph by K. A. Hobson. (Online version in colour.) 
Feather MeHg increased in both adult ( n 1⁄4 40) and first-winter ivory gulls ( n 1⁄4 40) from Arctic Canada and western Greenland between 1877 and 2007. White dots are first-winter birds; black dots are adult birds. 
Mercury (Hg) is increasing in marine food webs, especially at high latitudes. The bioaccumulation and biomagnification of methyl mercury (MeHg) has serious effects on wildlife, and is most evident in apex predators. The MeHg body burden in birds is the balance of ingestion and excretion, and MeHg in feathers is an effective indicator of overall MeHg burden. Ivory gulls (Pagophila eburnea), which consume ice-associated prey and scavenge marine mammal carcasses, have the highest egg Hg concentrations of any Arctic bird, and the species has declined by more than 80% since the 1980s in Canada. We used feathers from museum specimens from the Canadian Arctic and western Greenland to assess whether exposure to MeHg by ivory gulls increased from 1877 to 2007. Based on constant feather stable-isotope (δ(13)C, δ(15)N) values, there was no significant change in ivory gulls' diet over this period, but feather MeHg concentrations increased 45× (from 0.09 to 4.11 µg g(-1) in adults). This dramatic change in the absence of a dietary shift is clear evidence of the impact of anthropogenic Hg on this high-latitude threatened species. Bioavailable Hg is expected to increase in the Arctic, raising concern for continued population declines in high-latitude species that are far from sources of environmental contaminants. © 2015 The Author(s) Published by the Royal Society. All rights reserved.
Results of contour-detection experiments. (a) Contour detection as a function of element o¡set from alignment with a smooth curve, where contour elements consist of Gaussian-vignetted drifting sinusoids. O¡set dimension is scaled to the size of the elementsö¼ of the Gaussian vignette. The results of two observers are plotted; each datum plots the percentage correct and § 1 s.e.m. for 1000 two-alternate forced-choice trials; all conditions were interleaved in blocks of 50 trials. Observer L.M. was naive to the aims and manipulations of the experiment. The round symbols show detection performance as a function of o¡set of elements from a smooth curve, where the o¡set is in the opposite direction to the direction of drift of the sinusoid (i.e. o¡set is counter to motion-induced displacement). The square symbols show detection performance where the o¡set is in the same direction as the direction of drift of the sinusoid. These results indicate that the position of the elements that results in optimal contour-detection performance occurs when the elements are physically displaced from alignment along a smooth curve by about 1¼ for observer A.H. and 1.5¼ for observer L.M. (b) Contour detection as a function of element o¡set using Gaussian-vignetted counterphasing (i.e. non-drifting) sinusoids. The symbols show detection performance as a function of o¡set of contour elements from a smooth curve. As expected, performance simply deteriorates monotonically as o¡set increases, since no apparent displacement is induced by the counterphasing sinusoids. 
An assumption inherent in many models of visual space is that the spatial coordinates of retinal cells implicitly give rise to the perceptual code for position. The results of the experiments reported here, in which it is shown that retinally non-veridical locations of contour elements are used by the visual system for contour-element binding, lend support to a different view. The visual system does not implicitly code position with reference to the labelled locations of retinal cells, but dynamically extracts spatial position from the aggregate result of local computations. These computations may include local spatial relationships between retinal cells, but are not confined to them; other computations, including position derived from local velocity cues, are combined to code the position of objects in the visual world.
Aposematic insects use warning colours to deter predators, but many also produce odours or sounds when attacked by a predator. One possible role for these additional components is that they promote the association between the warning colour and the non-profitability it signals, thus reducing the chance of future attacks from visually hunting predators. This experiment explicitly tests this idea by looking at the effects of sound on a visual discrimination task. Young domestic chicks were trained to look for food rewards under coloured paper cones scattered in an experimental arena. In a subsequent visual discrimination task, they learned to discriminate between rewarded and non-rewarded hats on the basis of colour. Half the chicks performed this task in silence, whilst the other half had a tone played when they attacked non-rewarded hats. The presence of the tone improved the speed of colour discrimination learning. This demonstrates that there could be a selective advantage for aposematic coloured insects to emit sounds when attacked, since avian predators will learn to avoid their coloration more quickly. The role of psychological interactions between signal components in receivers is discussed in relation to the evolution of multimodal displays.
Location and size of PCR targets on the nuclear SSU and mitochondrial LSU rRNA genes from amber- and copal-preserved insects. PCR fragment sizes include primer sequences and are based on Drosophila melanogaster (nuclear) and Apis mellifera (mitochondrial) sequences. Upper case letters identify each target region amplified either in single or secondary \ nested PCR assays. Lower case letters identify primers listed in § 2. The location and size of PCR fragments amplified in previous studies of amber-preserved insects are shown for comparison. Fragment sizes are given as stated in the respective papers with the exception of the mitochondrial PCR targets of DeSalle et al . (1992), which were quoted as being 150 bp each. 
DNA sequences, and their frequency of occurrence, of cloned PCR product from amber- and copal- preserved bees. PCR products are identified by the specimen number and PCR target letter. Asterisks indicate PCR product amplified from extracts of fossil resin only (no insect tissue). Sequences 30 and 31 are composed entirely of tandem repeats of the forward or reverse primers b and f. Dots indicate identity with the first sequence. 
Phylogenetic relationships of DNA sequences from amber- and copal-preserved bees, inferred using the neighbour-joining algorithm. The trees were rooted using Saccharom ces cere isiae as an outgroup. Numbers adjacent to branches refer to bootstrap values (greater than 500) from 1000 replications. The three trees are based on sequences from the 84 bp, 139 bp, and 187 bp PCR fragments A, B and C. Asterisks indicate sequences obtained from extracts of fossil resin (no insect tissue). 
Apparently ancient DNA has been reported from amber-preserved insects many millions of years old. Rigorous attempts to reproduce these DNA sequences from amber- and copal-preserved bees and flies have failed to detect any authentic ancient insect DNA. Lack of reproducibility suggests that DNA does not survive over millions of years even in amber, the most promising of fossil environments.
Evolutionary and population dynamics models suggest that the migration rate will affect the probability of survival in fragmented landscapes. Using data for butterfly species in the fragmented British landscape and in immediately adjoining areas of the European continent, this paper shows that species of intermediate mobility have declined most, followed by those of low mobility, whereas high-mobility species are generally surviving well. Compared to the more sedentary species, species of intermediate mobility require relatively large areas where they breed at slightly lower local densities. Intermediate mobility species have probably fared badly through a combination of metapopulation (extinction and colonization) dynamics and the mortality of migrating individuals which fail to find new habitats in fragmented landscapes. Habitat fragmentation is likely to result in the non-random extinction of populations and species characterized by different levels of dispersal, although the details are likely to depend on the taxa, habitats and regions considered.
We have observed the changes in the intracellular ammonium (NH4+) content and the intracellular pH during administration of 20 mM NH4Cl (the ammonium pulse experiment) using nitrogen-14 and phosphorus-31 nuclear magnetic resonance spectroscopy (14N and 31P NMR) at 8.45 T. In the isolated perfused rat mandibular salivary gland, resonances of trimethylamines (-328 p.p.m.) and betaine (-329 p.p.m. from the resonance of NO3-) were detected. A chemical shift reagent, 10 mM of dysprosium triethylenetetramine-N,N,N',N",N"',N"'-hexaacetic acid (Dy(TTHA], was used to discriminate between the resonances from the extracellular NH4+ (-352 p.p.m.) and the intracellular NH4+ (-355 p.p.m.). During the NH4Cl application, the intracellular NH4+ content [( NH4+]i) increased quickly to ca. 50 mmol per litre intracellular fluid (ICF), then increased gradually to ca. 70 mmol per litre ICF. The intracellular pH (pHi), calculated from the 31P chemical shift of inorganic phosphate, increased transiently by 0.5 pH units and then decreased gradually in spite of the high level of [NH4+]i. The initial increase of [NH4+]i, which was observed by 14N NMR, was larger than that calculated from the intracellular pH on an assumption of a non-ionic diffusion process for ammonia. These results suggest a possibility of influx of NH4+, and also suggest an activation of cellular buffering mechanism that extrudes the excess bases from the cells.
Net speciation intervals (logarithmic model) in ® sh radiations plotted against age of radiations (logarithmic scale). Data from tables 1 and 2. The line is the best ® t to the data and is calculated excluding the Lake Victoria taxa. The inset is a magni® cation of the cluster of extremely recent radiations. Points that represent multiple observations have the number of independent observations attached. Filled points represent taxa from Lake Victoria. Aplocheilichthys and Clarias/Xenoclarias have been scored with SI lo g of 14 600 yr for the one in four lineages that must have given rise to the one endemic species in each of these two genera. Grey circles are the two extreme alternative placements of the haplochromine species ¯ ock of Lake Victoria obtained under the alternative assumptions of monophyly and polyphyly (see table 1). Except for the haplochromine species ¯ ock, all Lake Victoria taxa have speciation intervals on or above the line of best ® t for other ® sh radiations.  
Geophysical data are currently being interpreted as evidence for a late Pleistocene desiccation of Lake Victoria and its refilling 14,600 years ago. This implies that between 500 and 1000 endemic cichlid fish species must have evolved in 14,600 years, the fastest large-scale species radiation known. A recent review concludes that biological evidence clearly rejects the postulated Pleistocene desiccation of the lake: a 14,600 year history would imply exceptionally high speciation rates across a range of unrelated fish taxa. To test this suggestion, I calculated speciation rates for all 41 phylogenetic lineages of fish in the lake. Except for one cichlid lineage, accepting a 14 600 year history does not require any speciation rates that fall outside the range observed in fishes in other young lakes around the world. The exceptional taxon is a lineage of haplochromine cichlids that is also known for its rapid speciation elsewhere. Moreover, since it is unknown how many founding species it has, it is not certain that its speciation rates are really outside the range observed in fishes in other young lakes. Fish speciation rates are generally faster in younger than in older lakes, and those in Lake Victoria, by far the largest of the young lakes of the world, are no exception. From the speciation rates and from biogeographical observations that Lake Victoria endemics, which lack close relatives within the lake basin, have such relatives in adjacent drainage systems that may have had Holocene connections to Lake Victoria, I conclude that the composition of the fish assemblage does not provide biological evidence against Pleistocene desiccation. It supports a hypothesis of recent colonization from outside the lake basin rather than survival of a diverse assemblage within the basin.
Meta-analysis results. 
An essential requirement to determine a population's potential for evolutionary change is to quantify the amount of genetic variability expressed for traits under selection. Early investigations in laboratory conditions showed that the magnitude of the genetic and environmental components of phenotypic variation can change with environmental conditions. However, there is no consensus as to how the expression of genetic variation is sensitive to different environmental conditions. Recently, the study of quantitative genetics in the wild has been revitalized by new pedigree analyses based on restricted maximum likelihood, resulting in a number of studies investigating these questions in wild populations. Experimental manipulation of environmental quality in the wild, as well as the use of naturally occurring favourable or stressful environments, has broadened the treatment of different taxa and traits. Here, we conduct a meta-analysis on recent studies comparing heritability in favourable versus unfavourable conditions in non-domestic and non-laboratory animals. The results provide evidence for increased heritability in more favourable conditions, significantly so for morphometric traits but not for traits more closely related to fitness. We discuss how these results are explained by underlying changes in variance components, and how they represent a major step in our understanding of evolutionary processes in wild populations. We also show how these trends contrast with the prevailing view resulting mainly from laboratory experiments on Drosophila. Finally, we underline the importance of taking into account the environmental variation in models predicting quantitative trait evolution.
Primer pheromones are thought to act in a variety of vertebrates and invertebrates but only a few have been chemically identified. We report that a blend of ten fatty-acid esters found on the cuticles of honeybee larvae, already known as a kairomone, releaser pheromone and primer pheromone, also act as a primer pheromone in the regulation of division of labour among adult workers. Bees in colonies receiving brood pheromone initiated foraging at significantly older ages than did bees in control colonies in five out of five trials. Laboratory and additional field tests also showed that exposure to brood pheromone significantly depressed blood titres of juvenile hormone. Brood pheromone exerted more consistent effects on age at first foraging than on juvenile hormone, suggesting that the primer effects of this pheromone may occur via other, unknown, mechanisms besides juvenile hormone. These results bring the number of social factors known to influence honeybee division of labour to three: worker-worker interactions, queen mandibular pheromone and brood pheromone.
Per capita fecundity of fast (in red) and slow (in blue) selected S. ratti lines assayed in experiment 2 on days (a,b) 5 and (c,d ) 33 p.i. Open circles show the data and filled circles the model fit. 
Length of parasitic females from fast (diamonds) and slow (squares) selected S. ratti lines assayed in experiment 3. The error bars are standard errors and the dotted line is a cubic spline fitted to the data. 
Evolutionary ecology predicts that parasite life-history traits, including a parasite's survivorship and fecundity within a host, will evolve in response to selection and that their evolution will be constrained by trade-offs between traits. Here, we test these predictions using a nematode parasite of rats, Strongyloides ratti, as a model. We performed a selection experiment by passage of parasite progeny from either early in an infection ('fast' lines) or late in an infection ('slow' lines). We found that parasite fecundity responded to selection but that parasite survivorship did not. We found a trade-off mediated via conspecific density-dependent constraints; namely, that fast lines exhibit higher density-independent fecundity than slow lines, but fast lines suffered greater reduction in fecundity in the presence of density-dependent constraints than slow lines. We also found that slow lines both stimulate a higher level of IgG1, which is a marker for a Th2-type immune response, and show less of a reduction in fecundity in response to IgG1 levels than for fast lines. Our results confirm the general prediction that parasite life-history traits can evolve in response to selection and indicate that such evolutionary responses may have significant implications for the epidemiology of infectious disease.
Both appropriate metabolic rates and sufficient immune function are essential for survival. Consequently, eco-immunologists have hypothesized that animals may experience trade-offs between metabolic rates and immune function. Previous work has focused on how basal metabolic rate (BMR) may trade-off with immune function, but maximal metabolic rate (MMR), the upper limit to aerobic activity, might also trade-off with immune function. We used mice artificially selected for high mass-independent MMR to test for trade-offs with immune function. We assessed (i) innate immune function by quantifying cytokine production in response to injection with lipopolysaccharide and (ii) adaptive immune function by measuring antibody production in response to injection with keyhole limpet haemocyanin. Selection for high mass-independent MMR suppressed innate immune function, but not adaptive immune function. However, analyses at the individual level also indicate a negative correlation between MMR and adaptive immune function. By contrast BMR did not affect immune function. Evolutionarily, natural selection may favour increasing MMR to enhance aerobic performance and endurance, but the benefits of high MMR may be offset by impaired immune function. This result could be important in understanding the selective factors acting on the evolution of metabolic rates.
The refuge-high-dose strategy for delaying insect adaptation to transgenic plants produces non-transgenic plants that enable survival of susceptible individuals. Previous theoretical work has suggested three requirements for success of the refuge-high-dose strategy: a low initial frequency of the resistance allele, extensive mating between resistant and susceptible adults and recessive inheritance of resistance. In order to understand an observed decrease in resistance frequency and improve the potential for managing resistance better, we used analytical and simulation models for exploring the conditions that prevent or reverse the evolution of resistance, even when resistance is not rare initially. Assuming random mating and recessive or nearly recessive inheritance of resistance, the factors favouring reversal of resistance are non-recessive costs of resistance, low initial resistance allele frequency, large refuges, incomplete resistance and density-independent population growth in refuges.
The characteristics of the mariner transposable element in natural populations of Drosophila simulans from different parts of the world were analysed. The somatic excision rate (estimated from a test-cross with a reference strain), the average number of copies (determined by Southern blots), and the presence of deleted copies (detected by polymerase chain reaction amplification) were estimated for each population. There was a great variability in the somatic excision rate, measured as the percentage of mosaic males, both within and between populations. The population effect was highly significant. The average copy number also varied widely and was correlated with the excision rate. Rare deleted elements were detected by polymerase chain reaction and Southern blots. Percentage of mosaic males increased in strains kept for a long time at low temperature, and the somatic excision rate increased with the latitude of origin of populations. Therefore, these results strongly suggest that temperature could be involved in the regulation of mariner somatic excision in D. simulans.
Brachiopods and bivalves feed in similar ways and have occupied the same environments through geological time, but brachiopods were far more diverse and abundant in the Palaeozoic whereas bivalves dominate the post-Palaeozoic, suggesting a transition in ecological dominance 250 Ma. However, diversity and abundance data alone may not adequately describe key changes in ecosystem function, such as metabolic activity. Here, we use newly compiled body size data for 6066 genera of bivalves and brachiopods to calculate metabolic rates and revisit this question from the perspective of energy use, finding that bivalves already accounted for a larger share of metabolic activity in Palaeozoic oceans. We also find that the metabolic activity of bivalves has increased by more than two orders of magnitude over this interval, whereas brachiopod metabolic activity has declined by more than 50%. Consequently, the increase in bivalve energy metabolism must have occurred via the acquisition of new food resources rather than through the displacement of brachiopods. The canonical view of a mid-Phanerozoic transition from brachiopod to bivalve dominance results from a focus on taxonomic diversity and numerical abundance as measures of ecological importance. From a metabolic perspective, the oceans have always belonged to the clams.
Qualitative diagnosis of DWV using RT-PCR. In all gels, the arrow indicates the DWV product; M, marker; A, actin control. (a) DWV infection: lane 1-4, RT-PCR from different infected samples; lane 5, negative control (water). (b) Localization of DWV in body regions of honeybee: lane 1, head; lane 2, thorax; lane 3, abdomen; lane 4, DWV-positive control. (c) Oral application of DWV: lane 1, seventh day after continuous oral application of DWV lysate in sucrose; lane 2, seventh day after continuous oral application of control lysate in sucrose; lane 3, seventh day after continuous oral application of sucrose; lane 4, DWV-positive control; lane 5, negative control (water). (d ) Time course after DWV haemolymph injection: lane 1, DWV-positive control; lane 2-5 represent second to fifth day after DWV lysate injection, respectively; lane 6, negative control (water).
Sucrose responsiveness of bees was tested on the (a) first and (b) fourth day after artificial haemolymph infection with control lysate and DWV lysate. After each stimulation of an antenna with successive single stimuli of increasing sucrose concentrations (0–1 M), the PER (proboscis extension response) was recorded. The data show the mean responsiveness indicated by the percentage of PER. (First day, nZ60 bees each group and fourth day, nZ20 bees each group). The asterisks indicate significant difference between the groups (Fisher exact test; Ã p!0.05).  
In both mammals and invertebrates, virus infections can impair a broad spectrum of physiological functions including learning and memory formation. In contrast to the knowledge on the conserved mechanisms underlying learning, the effects of virus infection on different aspects of learning are barely known. We use the honeybee (Apis mellifera), a well-established model system for studying learning, to investigate the impact of deformed wing virus (DWV) on learning. Injection of DWV into the haemolymph of forager leads to a RT-PCR detectable DWV signal after 3 days. The detailed behavioural analysis of DWV-infected honeybees shows an increased responsiveness to water and low sucrose concentrations, an impaired associative learning and memory formation, but intact non-associative learning like sensitization and habituation. This contradicts all present studies in non-infected bees, where increased sucrose responsiveness is linked to improved associative learning and to changes in non-associative learning. Thus, DWV seems to interfere with molecular mechanism of learning by yet unknown processes that may include viral effects on the immune system and on gene expression.
For legend see opposite. 
Three-dimensional model of opsin molecule, derived from Baldwin (1993), showing the relative position of the seven α -helical transmembrane regions. The circles represent the seven α -helices, as if viewed from the cytosolic side of the molecule, and the lines within each circle indicate the orientation of amino acids within each helix. The length of each line represents the depth at which the amino acid is located within the helix. The positions of the candidate amino acid sites for spectral tuning, together with the site of chromophore attachment, are indicated by arrows. 
Percentage amino acid composition of the opsin proteins (The values presented are the percentage amino acid content, calculated in each case over the same part of the opsin protein. The total hydroxyl content is a sum of the percentage content of serine, threonine and tyrosine.)
Neighbour-joining tree for fish rod opsin gene sequences. The bootstrap confidence values are shown for each branch. The length of each branch reflects the extent of nucleotide divergence. The scale bar is calibrated in substitutions per site. The lamprey ( Lampetra japonica ), goldfish ( arassius auratus ), carp ( rinus carpio ), cottoid ( Asprocottus korotneffi ), sand goby ( P . minutus ), eel ( A . anguilla ) and cave fish ( Ast anax fasciatus ) sequences are from Hisatomi et al . (1991), Johnson et al . (1993), Tsai et al . (1995), Hunt et al . (1996), Archer et al . (1992), Archer et al . (1996) and Yokoyama et al . (1995), respectively. 
The main object of this study was to investigate the molecular basis for changes in the spectral sensitivity of the visual pigments of deep-sea fishes. The four teleost species studied, Hoplostethus mediterraneus, Cataetyx laticeps, Gonostoma elongatum and Histiobranchus bathybius, are phylogenetically distant from each other and live at depths ranging from 500 to almost 5000 m. A single fragment of the intronless rod opsin gene was PCR-amplified from each fish and sequenced. The wavelength of peak sensitivity for the rod visual pigments of the four deep-sea species varies from 483 nm in H. mediterraneus and G. elongatum to 468 nm in C. laticeps. Six amino acids at sites on the inner face of the chromophore-binding pocket formed by the seven transmembrane a-helices are identified as candidates for spectral tuning. Substitutions at these sites involve either a change of charge, or a gain or loss of a hydroxyl group. Two of these, at positions 83 and 292, are consistently substituted in the visual pigments of all four species and are likely to be responsible for the shortwave sensitivity of the pigments. Shifts to wavelengths shorter than 480 nm may involve substitution at one or more of the remaining four sites. None of the modifications found in the derived sequences of these opsins suggest functional adaptations, such as increased content of hydroxyl-bearing or proline residues, to resist denaturation by the elevated hydrostatic pressures of the deep sea. Phylogenetic evidence for the duplication of the rod opsin gene in the Anguilliform lineage is presented.
Encapsulation ability of control lines (open symbols) and lines selected for defence against L. boulardi (solid symbols). Selection was halted at generation 9. 
Competitive ability of control (open bars) and selection (black bars) lines at four levels of food (*p approximately 0.05, **p50.01; see text for further details of statistics). 
Costs of resistance are widely assumed to be important in the evolution of parasite and pathogen defence in animals, but they have been demonstrated experimentally on very few occasions. Endoparasitoids are insects whose larvae develop inside the bodies of other insects where they defend themselves from attack by their hosts' immune systems (especially cellular encapsulation). Working with Drosophila melanogaster and its endoparasitoid Leptopilina boulardi, we selected for increased resistance in four replicate populations of flies. The percentage of flies surviving attack increased from about 0.5% to between 40% and 50% in five generations, revealing substantial additive genetic variation in resistance in the field population from which our culture was established. In comparison with four control lines, flies from selected lines suffered from lower larval survival under conditions of moderate to severe intraspecific competition.
Study site. The three fossil deposits, PV (42858 0 22.0 00 S, 172835 0 49.0 00 E), BHV (42858 0 19.36 00 S, 172839 0 56.15 00 E) and Rosslea (42857 0 53.83 00 S, 172839 0 22.39 00 E), from which 158 radiocarbon-dated moa fossils were characterized for DNA decay kinetics. The sites in North Canterbury, South Island, New Zealand are located within a 5 km radius in the eastern rain shadows of the Southern Alps. Most of the area is more than 200 m a.s.l. and consists of flat alluvial plains and rolling downlands. Information on calibrated radiocarbon ages and DNA preservation (C T values) are shown for each site. m, mean age. 
Observed and predicted rates of DNA decay. The predicted survival of DNA in bone through time, measured as intact phosphodiester bonds in the DNA backbone (a), and survival of a 242 bp fragment (b). The depicted survival rates are based on: (i) the average mtDNA decay rate measured directly from qPCR of 158 moa bones; (ii) the rate of depurination measured from DNA in solution at pH 5 in Lindahl & Nyberg [22] but adjusted to 13.18C to allow comparison with the moa data; (iii) the same rate adjusted further to pH 7.5, as expected inside a bone; (iv) mtDNA and nuDNA decay rates calculated based on Illumina HiSeq data from two moa samples (HiSeq 1 from sample S40114, HiSeq 2 from sample S39946-3). The estimated decay rate (k, per site per year) is listed for each of the seven datasets. 
Claims of extreme survival of DNA have emphasized the need for reliable models of DNA degradation through time. By analysing mitochondrial DNA (mtDNA) from 158 radiocarbon-dated bones of the extinct New Zealand moa, we confirm empirically a long-hypothesized exponential decay relationship. The average DNA half-life within this geographically constrained fossil assemblage was estimated to be 521 years for a 242 bp mtDNA sequence, corresponding to a per nucleotide fragmentation rate (k) of 5.50 × 10(-6) per year. With an effective burial temperature of 13.1°C, the rate is almost 400 times slower than predicted from published kinetic data of in vitro DNA depurination at pH 5. Although best described by an exponential model (R(2) = 0.39), considerable sample-to-sample variance in DNA preservation could not be accounted for by geologic age. This variation likely derives from differences in taphonomy and bone diagenesis, which have confounded previous, less spatially constrained attempts to study DNA decay kinetics. Lastly, by calculating DNA fragmentation rates on Illumina HiSeq data, we show that nuclear DNA has degraded at least twice as fast as mtDNA. These results provide a baseline for predicting long-term DNA survival in bone.
Top-cited authors
Marcel E Visser
  • Netherlands Institute of Ecology (NIOO-KNAW)
Locke Rowe
  • University of Toronto
Neil B Metcalfe
  • University of Glasgow
Teja Tscharntke
  • Georg-August-Universität Göttingen
Nicholas K Dulvy
  • Simon Fraser University