Ecological and evolutionary drivers of range size in Coenagrion damselflies.

Geographic range size is a key ecological and evolutionary characteristic of a species, yet the causal basis of variation in range size among species remains largely unresolved. One major reason for this is that several ecological and evolutionary traits may jointly shape species' differences in range size. We here present an integrated study of the contribution of ecological (dispersal capacity, body size and latitudinal position) and macroevolutionary (species' age) traits in shaping variation in species' range size in Coenagrion damselflies. We reconstructed the phylogenetic tree of this genus to account for evolutionary history when assessing the contribution of the ecological traits and to evaluate the role of the macroevolutionary trait (species' age). The genus invaded the Nearctic twice independently from the Palearctic, yet this was not associated with the evolution of larger range sizes or dispersal capacity. Body size and species' age did not explain variation in range size. There is higher flight ability (as measured by wing aspect ratio) at higher latitudes. Species with a larger wing aspect ratio had a larger range size, also after correcting for phylogeny, suggesting a role for dispersal capacity in shaping the species' ranges. More northern species had a larger species' range, consistent with Rapoport's rule, possibly related to niche width. Our results underscore the importance of integrating macroecology and macroevolution when explaining range size variation among species.

Movement patterns and genetic diversity of wild and reintroduced common dormice, Muscardinus avellanarius.

Movement is an important life history trait that can have an impact on local adaptation, and other evolutionary phenomena. We used a combination of nestbox survey data and genetic techniques (genotyping at 10 microsatellite loci) to quantify patterns of movement in common dormice Muscardinus avellanarius at two distinct sites in the UK: 1) Bontuchel (a natural population) and 2) Wych (captive-bred individuals that were reintroduced to this site), over three consecutive years (2006-2008). Both methods revealed a consistent pattern of sex-biased movement (movements by adult males and females) in both populations that allowed isolation-by-distance genetic structure to develop within 1 km. The similarity of data from captive-bred and natural individuals indicated that ex situ programing has not significantly altered the natural movement behavior of common dormice; consequently, the two populations could be managed with the same conservation strategies. We also found that the reintroduced dormice in Wych maintained relatively high levels of genetic diversity. This first report of movement patterns in reintroduced and natural populations of M. avellanarius combining genetic and field-survey data highlights the role of genetic studies in the investigation of ecological behaviour and for conservation management.

Persistent genetic signatures of historic climatic events in an Antarctic octopus.

Repeated cycles of glaciation have had major impacts on the distribution of genetic diversity of the Antarctic marine fauna. During glacial periods, ice cover limited the amount of benthic habitat on the continental shelf. Conversely, more habitat and possibly altered seaways were available during interglacials when the ice receded and the sea level was higher. We used microsatellites and partial sequences of the mitochondrial cytochrome oxidase 1 gene to examine genetic structure in the direct-developing, endemic Southern Ocean octopod Pareledone turqueti sampled from a broad range of areas that circumvent Antarctica. We find that, unusually for a species with poor dispersal potential, P. turqueti has a circumpolar distribution and is also found off the islands of South Georgia and Shag Rocks. The overriding pattern of spatial genetic structure can be explained by hydrographic (with ocean currents both facilitating and hindering gene flow) and bathymetric features. The Antarctic Peninsula region displays a complex population structure, consistent with its varied topographic and oceanographic influences. Genetic similarities between the Ross and Weddell Seas, however, are interpreted as a persistent historic genetic signature of connectivity during the hypothesized Pleistocene West Antarctic Ice Sheet collapses. A calibrated molecular clock indicates two major lineages within P. turqueti, a continental lineage and a sub-Antarctic lineage, that diverged in the mid-Pliocene with no subsequent gene flow. Both lineages survived subsequent major glacial cycles. Our data are indicative of potential refugia at Shag Rocks and South Georgia and also around the Antarctic continent within the Ross Sea, Weddell Sea and off Adélie Land. The mean age of mtDNA diversity within these main continental lineages coincides with Pleistocene glacial cycles.

Developmental plasticity as a cohesive evolutionary process between sympatric alternate-year insect cohorts.

Many species, particularly insects, pass through a series of distinct phases during their life history, with the developmental timing directed towards appropriate resources. Any factor that creates variation in developmental timing may partition a population into discrete populations-or 'cohorts'. Where there is continued failure to recruit outside the natal cohort then alternate cohorts will have their own internal dynamics, eventually leading to independent demographic and evolutionary trajectories. By contrast, continued variation in development rates within a cohort-cohort splitting-may homogenise otherwise independent demographic units. Using a panel of 14 microsatellite loci, we quantify the genetic signature of apparent demographic isolation between coexisting, but alternate, semivoltine cohorts of the damselfly Coenagrion mercuriale at locations that span its distribution in the UK. We find consistently low levels of genetic divergence between sympatric cohorts of C. mercuriale, indicative of developmental plasticity during the larval stage (unregulated development) whereby some individuals complete their development outside the predominant 2-year (semivoltine) period. Thus, individuals that alter their developmental rate successfully recruit to a different cohort. Despite maintaining contrasting population sizes, gene flow between alternate cohorts broadly is sufficient to place them on a similar evolutionary trajectory and also buffers against loss of genetic diversity. Such flexible larval development permits a response to local conditions and may facilitate response to environmental change.

Empirical evidence of senescence in adult damselflies (Odonata: Zygoptera).

1. Age-dependent increases in mortality have been documented in a variety of species of insect under laboratory conditions. However, while strong statistical evidence has been presented for senescence in vertebrate populations in the wild, we know little about the rate and shape of senescence in wild populations of insects. 2. Odonates (damselflies and dragonflies) provide excellent candidate species for evaluating demographic senescence as they are large enough to be marked individually and they are easily re-sighted without recapture. The prevailing opinion - based entirely on qualitative examination of the declines in log numbers alive with time since marking - is that odonates exhibit age-independent daily survivorship. 3. Here, we examine mark-recapture data on the Azure Damselfly Coenagrion puella over two consecutive seasons. For the first time, we evaluate and compare the fit of quantitative models that not only account for weather-dependent daily variation in daily re-sighting rates, but also age-dependent variation in daily survivorship. 4. Models with age-dependent declines in daily survivorship provide a more parsimonious explanation for the data than similar models without these age-dependent effects. In general, models in which mortality increases in an exponential (Gompertz) fashion explain the mark-recapture sequences more efficiently than a range of alternative models, including those in which mortality increases as a power function (Weibull) or reaches a plateau (logistic). These results are indicative of a general senescent decline in physiological functioning, which is particularly marked after 15 days as a mature adult. 5. Weather (temperature, sun and precipitation) and initial mite load influenced the probability of daily re-sighting. Weather and mite load also influenced daily survivorship, but their effects differed between seasons. 6. Overall, fitting models with age as an explicit covariate demonstrates that odonates do indeed senesce. This contradicts previously held assumptions that Odonata do not exhibit age-dependent survivorship in the wild.

A panel of ten microsatellite loci for the Chagas disease vector Rhodnius prolixus (Hemiptera: Reduviidae).

Rhodnius prolixus is the main vector of Chagas disease in Venezuela, where it is found colonising rural housing consisting of unplastered adobe walls with palm and/or metal roofs. Vector control failure in Venezuela may be due to the invasion of houses by silvatic populations of R. prolixus found in palms. As part of a study to determine if domestic and silvatic populations of R. prolixus are isolated, thus clarifying the role of silvatic populations in maintaining house infestations, we constructed three partial genomic microsatellite libraries. A panel of ten dinucleotide polymorphic microsatellite markers was selected for genotyping. Allele numbers per locus ranged from three to twelve, with observed and expected heterozygosity ranging from 0.26 to 0.55 and 0.32 to 0.66. The microsatellite markers presented here will contribute to the control of Chagas disease in Venezuela and Colombia through the provision of population information that may allow the design of improved control strategies.

Poly(3- hydroxybutyrate)/Bioglass(®) composite films containing carbon nanotubes.

Poly(3hydroxybutyrate) (P(3HB))/Bioglass(®) composites incorporating multiwalled carbon nanotubes (MWCNTs) have been successfully prepared by the solvent casting technique. The microstructure, electrical properties and bioactivity of the composites were characterized using scanning electron microscopy, x-ray diffraction and current-voltage measurements. Different concentrations of MWCNTs were used to determine their effect on the electrical properties of the composites. MWCNTs and Bioglass(®) particles were found to be homogeneously dispersed throughout the P(3HB) matrix. The electrical resistance of the composite samples decreased on increasing the MWCNT concentration, as expected. An in vitro degradation study in simulated body fluid (SBF) was carried out on composite samples. The formation of hydroxyapatite on the surfaces of P(3HB)/Bioglass(®)/MWCNT composite films was confirmed after two months of immersion in SBF. This hydroxyapatite layer was not formed on the neat polymeric films and on composites containing MWCNTs only (without Bioglass(®)). It was found that the presence of MWCNTs did not hinder the bioactivity of the Bioglass(®) particles, as confirmed by SEM and XRD studies on composite samples.

Leishmania donovani is the only cause of visceral leishmaniasis in East Africa; previous descriptions of L. infantum and "L. archibaldi" from this region are a consequence of convergent evolution in the isoenzyme data.

Isoenzyme-based studies have identified 3 taxa/species/'phylogenetic complexes' as agents of visceral leishmaniasis in Sudan: L. donovani, L. infantum and "L. archibaldi". However, these observations remain controversial. A new chitinase gene phylogeny was constructed in which stocks of all 3 putative species isolated in Sudan formed a monophyletic clade. In order to construct a more robust classification of the L. donovani complex, a panel of 16 microsatellite markers was used to describe 39 stocks of these 3 species. All "L. donovani complex" stocks from Sudan were again found to form a single monophyletic clade. L. donovani ss stocks from India and Kenya were found to form 2 region-specific clades. The partial sequence of the glutamate oxaloacetate transaminase (GOT) gene of 17 L. donovani complex stocks was obtained. A single nucleotide polymorphism in the GOT gene appeared to underlie the isoenzyme classification. It was concluded that isoenzyme-based identification is unsafe for stocks isolated in L. donovani endemic areas and identified as L. infantum. It was also concluded that the name L. archibaldi is invalid and that only a single visceralizing species, Leishmania donovani, is found in East Africa.

Molecular and ecological evidence for small-scale isolation by distance in an endangered damselfly, Coenagrion mercuriale.

Coenagrion mercuriale (Charpentier) (Odonata: Zygoptera) is one of Europe's most threatened damselflies and is listed in the European Habitats directive. We combined an intensive mark-release-recapture (MRR) study with a microsatellite-based genetic analysis for C. mercuriale from the Itchen Valley, UK, as part of an effort to understand the dispersal characteristics of this protected species. MRR data indicate that adult damselflies are highly sedentary, with only a low frequency of interpatch movement that is predominantly to neighbouring sites. This restricted dispersal leads to significant genetic differentiation throughout most of the Itchen Valley, except between areas of continuous habitat, and isolation by distance (IBD), even though the core populations are separated by less than 10 km. An urban area separating some sites had a strong effect on the spatial genetic structure. Average pairwise relatedness between individual damselflies is positive at short distances, reflecting fine-scale genetic clustering and IBD both within- and between-habitat patches. Damselflies from a fragmented habitat have higher average kinship than those from a large continuous population, probably because of poorer dispersal and localized breeding in the former. Although indirect estimates of gene flow must be interpreted with caution, it is encouraging that our results indicate that the spatial pattern of genetic variation matches closely with that expected from direct observations of movement. These data are further discussed with respect to possible barriers to dispersal within the study site and the ecology and conservation of C. mercuriale. To our knowledge, this is the first report of fine-scale genetic structuring in any zygopteran species.

A possible route to large-scale production of SWNTs through a combination of the substrate and floating catalyst methods.

We describe a possible route for large-scale synthesis of single wall carbon nanotubes (SWNTs) by a combination of the substrate and floating methods. The template prohibits metal particle aggregation, resulting in high-purity SWNT growth, and the three-dimensional floating seeded state of the template induces improved yields of SWNTs in a semi-continuous system. The SWNTs obtained by this method exhibit large variations of texture (isolated tubes and bundles) and a wide range of diameters (0.4-4 nm).

Genetic identification of two sibling species of Lutzomyia longipalpis (Diptera: Psychodidae) that produce distinct male sex pheromones in Sobral, Ceará State, Brazil.

Lutzomyia longipalpis, the main sandfly vector for New World visceral leishmaniasis is a complex of an as yet undefined number of sibling species. At present, there is no consensus on the status (single species vs. species complex) of Brazilian populations. We applied five microsatellite loci to test the hypothesis that L. longipalpis occurs as two sympatric cryptic species in Sobral, Ceará State, Brazil as predicted by male sex pheromone chemotypes described previously for field specimens from this site [S-9-methyl-germacrene-B (9MGB) and a cembrene compound]. Abdominal spot morphology corresponds with pheromone type at this locality (9MGB in '1 spot' males and cembrene in '2 spot' males). Genotype data from 190 wild-caught L. longipalpis specimens collected in October 1999 and April 2001 were used to estimate genetic differentiation between the two sex pheromone populations and sampling dates. No significant (P > 0.05) genetic differences were found between the 1999 and 2001 9MGB samples (theta = 0.018; RST = -0.005), and genetic differentiation was low between the cembrene collections (theta = 0.037, P < 0.05; RST = -0.043, P > 0.05). By contrast, highly divergent allelic frequencies (largely at two microsatellite loci) corresponded to significant (P > 0.05) genetic differentiation (theta = 0.221; RST = 0.215) for all comparisons between samples with different pheromones. When pheromone samples were pooled across sample date, genetic differentiation was high (theta = 0.229; P < 0.001; Nem = 0.84). The allele frequency distribution at each of the five microsatellite loci was similar for males and females from the two collection years. Two of these loci showed highly divergent allele frequencies in the two sex pheromone populations. This was reflected in the highly significant genetic differentiation obtained from the male genotypes, between populations producing different pheromones (theta = 0.229-0.268; P < 0.0001 for the 2001 and theta = 0.254-0.558; P < 0.0001 for the 1999 collections, respectively). Similar results were obtained when the females, assigned to a pheromone type, were included in the analysis. Both a Bayesian analysis of the data set and a population assignment test provided strong evidence for two distinct populations corresponding to pheromone type. Given its genotype, the probability of assigning a 9MGB male to the original 9MGB population was 100% once the two years' collections were pooled. For cembrene-producing '2 spot' males this probability although still high, was lower than for 9MGB males, at 86%. This microsatellite data together with previously reported reproductive isolation between the two Sobral populations confirm that premating barriers are important in speciation of L. longipalpis.

CP-225,917 and CP-263,114, novel Ras farnesylation inhibitors from an unidentified fungus. I. Taxonomy, fermentation, isolation, and biochemical properties.

During the course of our screening for squalene synthase inhibitors and Ras farnesylation inhibitors, a novel fungal culture was discovered to produce two structurally unique compounds, CP-225,917 and CP-263,114, as well as zaragozic acid A (squalestatin I). The two compounds are characterized by a bicyclo[4.3.1]dec-1,6-diene core plus two extended alkyl chains. CP-225,917 and CP-263,114 inhibit Ras farnesyl transferase from rat brain with IC50 values of 6 microM and 20 microM, respectively. CP-225,917 inhibits squalene synthase with an IC50 value of 43 microM and CP-263,114 with an IC50 of 160 microM. The producing organism, though not fully classified, exhibits the characteristics of a sterile Phoma species.

Structure of the natural antibiotic ionophore CP-54,883.

The structural formula of the novel antibiotic ionophore CP-54,883 is deduced by systematic reduction from its 13C and 1H NMR spectra. The molecule consists of a polyether ring network and side chain terminated by an aromatic ring containing a phenoxy and two chlorine substituents. Based partly on an assumed analogy to corresponding regions of the similar structure nigericin-A1, the configurations about the sixteen asymmetric carbons are also derived. A belated crystal structure determination confirms, with the exception of one configuration assumed from nigericin-A1, the conclusions drawn, and shows that the anionic charge is in the phenoxy group, rather than the carboxylic acid function.

CP-54,883 a novel chlorine-containing polyether antibiotic produced by a new species of Actinomadura: taxonomy of the producing culture, fermentation, physico-chemical and biological properties of the antibiotic.

The novel chlorine-containing acidic polycyclic ether antibiotic CP-54,883 (C41H62O12Cl2) [corrected] is produced by the fermentation of Actinomadura routienii Huang sp. nov. This report presents the taxonomy and the fermentation conditions for the antibiotic-producing culture. The antibiotic is mainly active against Gram-positive bacteria. It protects chickens against Eimeria challenge in vivo and enhances rumen propionic acid in vitro. The physico-chemical properties are also characterized.