Within-host evolution of Staphylococcus aureus during asymptomatic carriage

Background Staphylococcus aureus is a major cause of healthcare associated mortality, but like many important bacterial pathogens, it is a common constituent of the normal human body flora. Around a third of healthy adults are carriers. Recent evidence suggests that evolution of S. aureus during nasal carriage may be associated with progression to invasive disease. However, a more detailed understanding of within-host evolution under natural conditions is required to appreciate the evolutionary and mechanistic reasons why commensal bacteria such as S. aureus cause disease. Therefore we examined in detail the evolutionary dynamics of normal, asymptomatic carriage. Sequencing a total of 131 genomes across 13 singly colonized hosts using the Illumina platform, we investigated diversity, selection, population dynamics and transmission during the short-term evolution of S. aureus. Principal Findings We characterized the processes by which the raw material for evolution is generated: micro-mutation (point mutation and small insertions/deletions), macro-mutation (large insertions/deletions) and the loss or acquisition of mobile elements (plasmids and bacteriophages). Through an analysis of synonymous, non-synonymous and intergenic mutations we discovered a fitness landscape dominated by purifying selection, with rare examples of adaptive change in genes encoding surface-anchored proteins and an enterotoxin. We found evidence for dramatic, hundred-fold fluctuations in the size of the within-host population over time, which we related to the cycle of colonization and clearance. Using a newly-developed population genetics approach to detect recent transmission among hosts, we revealed evidence for recent transmission between some of our subjects, including a husband and wife both carrying populations of methicillin-resistant S. aureus (MRSA). Significance This investigation begins to paint a picture of the within-host evolution of an important bacterial pathogen during its prevailing natural state, asymptomatic carriage. These results also have wider significance as a benchmark for future systematic studies of evolution during invasive S. aureus disease.

Genotypic and phenotypic diversity of a baculovirus population within an individual insect host

It is becoming increasingly apparent that many pathogen populations, including those of insects, show high levels of genotypic variation. Baculoviruses are known to be highly variable, with isolates collected from the same species in different geographical locations frequently showing genetic variation and differences in their biology. More recent Studies at smaller scales have also shown that virus DNA profiles from individual larvae can show polymorphisms within and between populations of the same species. Here, we investigate the genotypic and phenotypic variation of an insect baculovirus infection within a single insect host. Twenty four genotypically distinct nucleopolyhedrovirus (NPV) variants were isolated from an individual pine beauty moth, Panolis flammea, caterpillar by in vivo cloning techniques. No variant appeared to be dominant in the population. The Pafl NPV variants have been mapped using three restriction endonucleases and shown to contain three hypervariable regions containing insertions of 70-750 bp. Comparison of seven of these variants in an alternative host, Mamestra brassicae, demonstrated that the variants differed significantly in both pathogenicity and speed of kill. The generation and maintenance of pathogen heterogeneity are discussed. (c) 2005 Elsevier Inc. All rights reserved.

Linking the coevolutionary and population dynamics of host-parasitoid interactions

The interplay between coevolutionary and population or community dynamics is currently the focus of much empirical and theoretical consideration. Here, we develop a simulation model to study the coevolutionary and population dynamics of a hypothetical host-parasitoid interaction. In the model, host resistance and parasitoid virulence are allowed to coevolve. We investigate how trade-offs associated with these traits modify the system's coevolutionary and population dynamics. The most important influence on these dynamics comes from the incorporation of density-dependent costs of resistance ability. We find three main outcomes. First, if the costs of resistance are high, then one or both of the players go extinct. Second, when the costs of resistance are intermediate to low, cycling population and coevolutionary dynamics are found, with slower evolutionary changes observed when the costs of virulence are also low. Third, when the costs associated with resistance and virulence are both high, the hosts trade-off resistance against fecundity and invest little in resistance. However, the parasitoids continue to invest in virulence, leading to stable host and parasitoid population sizes. These results support the hypothesis that costs associated with resistance and virulence will maintain the heritable variation in these traits found in natural populations and that the nature of these trade-offs will greatly influence the population dynamics of the interacting species.

Evolutionary dynamics of Staphylococcus aureus during progression from carriage to disease

Whole-genome sequencing offers new insights into the evolution of bacterial pathogens and the etiology of bacterial disease. Staph- ylococcus aureus is a major cause of bacteria-associated mortality and invasive disease and is carried asymptomatically by 27% of adults. Eighty percent of bacteremias match the carried strain. How- ever, the role of evolutionary change in the pathogen during the progression from carriage to disease is incompletely understood. Here we use high-throughput genome sequencing to discover the genetic changes that accompany the transition from nasal carriage to fatal bloodstream infection in an individual colonized with meth- icillin-sensitive S. aureus. We found a single, cohesive population exhibiting a repertoire of 30 single-nucleotide polymorphisms and four insertion/deletion variants. Mutations accumulated at a steady rate over a 13-mo period, except for a cluster of mutations preceding the transition to disease. Although bloodstream bacteria differed by just eight mutations from the original nasally carried bacteria, half of those mutations caused truncation of proteins, including a prema- ture stop codon in an AraC-family transcriptional regulator that has been implicated in pathogenicity. Comparison with evolution in two asymptomatic carriers supported the conclusion that clusters of pro- tein-truncating mutations are highly unusual. Our results demon- strate that bacterial diversity in vivo is limited but nonetheless detectable by whole-genome sequencing, enabling the study of evolutionary dynamics within the host. Regulatory or structural changes that occur during carriage may be functionally important for pathogenesis; therefore identifying those changes is a crucial step in understanding the biological causes of invasive bacterial disease.

Dynamics of convectively driven banded jets in the laboratory

The banded organization of clouds and zonal winds in the atmospheres of the outer planets has long fascinated observers. Several recent studies in the theory and idealized modeling of geostrophic turbulence have suggested possible explanations for the emergence of such organized patterns, typically involving highly anisotropic exchanges of kinetic energy and vorticity within the dissipationless inertial ranges of turbulent flows dominated (at least at large scales) by ensembles of propagating Rossby waves. The results from an attempt to reproduce such conditions in the laboratory are presented here. Achievement of a distinct inertial range turns out to require an experiment on the largest feasible scale. Deep, rotating convection on small horizontal scales was induced by gently and continuously spraying dense, salty water onto the free surface of the 13-m-diameter cylindrical tank on the Coriolis platform in Grenoble, France. A “planetary vorticity gradient” or “β effect” was obtained by use of a conically sloping bottom and the whole tank rotated at angular speeds up to 0.15 rad s−1. Over a period of several hours, a highly barotropic, zonally banded large-scale flow pattern was seen to emerge with up to 5–6 narrow, alternating, zonally aligned jets across the tank, indicating the development of an anisotropic field of geostrophic turbulence. Using particle image velocimetry (PIV) techniques, zonal jets are shown to have arisen from nonlinear interactions between barotropic eddies on a scale comparable to either a Rhines or “frictional” wavelength, which scales roughly as (β/Urms)−1/2. This resulted in an anisotropic kinetic energy spectrum with a significantly steeper slope with wavenumber k for the zonal flow than for the nonzonal eddies, which largely follows the classical Kolmogorov k−5/3 inertial range. Potential vorticity fields show evidence of Rossby wave breaking and the presence of a “hyperstaircase” with radius, indicating instantaneous flows that are supercritical with respect to the Rayleigh–Kuo instability criterion and in a state of “barotropic adjustment.” The implications of these results are discussed in light of zonal jets observed in planetary atmospheres and, most recently, in the terrestrial oceans.

Water vapor variability in the tropics and its links to dynamics and precipitation

The distribution and variability of water vapor and its links with radiative cooling and latent heating via precipitation are crucial to understanding feedbacks and processes operating within the climate system. Column-integrated water vapor (CWV) and additional variables from the European Centre for Medium-Range Weather Forecasts (ECMWF) 40-year reanalysis (ERA40) are utilized to quantify the spatial and temporal variability in tropical water vapor over the period 1979–2001. The moisture variability is partitioned between dynamical and thermodynamic influences and compared with variations in precipitation provided by the Climate Prediction Center Merged Analysis of Precipitation (CMAP) and the Global Precipitation Climatology Project (GPCP). The spatial distribution of CWV is strongly determined by thermodynamic constraints. Spatial variability in CWV is dominated by changes in the large-scale dynamics, in particular associated with the El Niño–Southern Oscillation (ENSO). Trends in CWV are also dominated by dynamics rather than thermodynamics over the period considered. However, increases in CWV associated with changes in temperature are significant over the equatorial east Pacific when analyzing interannual variability and over the north and northwest Pacific when analyzing trends. Significant positive trends in CWV tend to predominate over the oceans while negative trends in CWV are found over equatorial Africa and Brazil. Links between changes in CWV and vertical motion fields are identified over these regions and also the equatorial Atlantic. However, trends in precipitation are generally incoherent and show little association with the CWV trends. This may in part reflect the inadequacies of the precipitation data sets and reanalysis products when analyzing decadal variability. Though the dynamic component of CWV is a major factor in determining precipitation variability in the tropics, in some regions/seasons the thermodynamic component cancels its effect on precipitation variability.

Tropospheric planetary wave dynamics and mixture modeling: Two preferred regimes and a regime shift

Investigation of preferred structures of planetary wave dynamics is addressed using multivariate Gaussian mixture models. The number of components in the mixture is obtained using order statistics of the mixing proportions, hence avoiding previous difficulties related to sample sizes and independence issues. The method is first applied to a few low-order stochastic dynamical systems and data from a general circulation model. The method is next applied to winter daily 500-hPa heights from 1949 to 2003 over the Northern Hemisphere. A spatial clustering algorithm is first applied to the leading two principal components (PCs) and shows significant clustering. The clustering is particularly robust for the first half of the record and less for the second half. The mixture model is then used to identify the clusters. Two highly significant extratropical planetary-scale preferred structures are obtained within the first two to four EOF state space. The first pattern shows a Pacific-North American (PNA) pattern and a negative North Atlantic Oscillation (NAO), and the second pattern is nearly opposite to the first one. It is also observed that some subspaces show multivariate Gaussianity, compatible with linearity, whereas others show multivariate non-Gaussianity. The same analysis is also applied to two subperiods, before and after 1978, and shows a similar regime behavior, with a slight stronger support for the first subperiod. In addition a significant regime shift is also observed between the two periods as well as a change in the shape of the distribution. The patterns associated with the regime shifts reflect essentially a PNA pattern and an NAO pattern consistent with the observed global warming effect on climate and the observed shift in sea surface temperature around the mid-1970s.

Assessing nitrogen dynamics in European ecosystems, integrating measurement and modelling: conclusions

This contribution closes this special issue of Hydrology and Earth System Sciences concerning the assessment of nitrogen dynamics in catchments across Europe within a semi-distributed Integrated Nitrogen model for multiple source assessment in Catchments (INCA). New developments in the understanding of the factors and processes determining the concentrations and loads of nitrogen are outlined. The ability of the INCA model to simulate the hydrological and nitrogen dynamics of different European ecosystems is assessed and the results of the first scenario analyses investigating the impacts of deposition, climatic and land-use change on the nitrogen dynamics are summarised. Consideration is given as to how well the model has performed as a generic too] for describing the nitrogen dynamics of European ecosystems across Arctic, Maritime. Continental and Mediterranean climates, its role in new research initiatives and future research requirements.

Simulating the spatial variation of soil mineral N within fields using the model SUNDIAL

Variable rate applications of nitrogen (N) are of environmental and economic interest. Regular measurements of soil N supply are difficult to achieve practically. Therefore accurate model simulations of soil N supply might provide a practical solution for site-specific management of N. Mineral N, an estimate of N supply, was simulated by the model SUNDIAL (Simulation of Nitrogen Dynamics In Arable Land) at more than 100 locations within three arable fields in Bedfordshire, UK. The results were compared with actual measurements. The outcomes showed that the spatial patterns of the simulations of mineral N corresponded to the measurements but the range of values was underestimated.

Modelling stream and soil water nitrate dynamics during experimentally increased nitrogen deposition in a coniferous forest catchment at Gardsjon, Sweden

Increased atmospheric deposition of inorganic nitrogen (N) may lead to increased leaching of nitrate (NO3-) to surface waters. The mechanisms responsible for, and controls on, this leaching are matters of debate. An experimental N addition has been conducted at Gardsjon, Sweden to determine the magnitude and identify the mechanisms of N leaching from forested catchments within the EU funded project NITREX. The ability of INCA-N, a simple process-based model of catchment N dynamics, to simulate catchment-scale inorganic N dynamics in soil and stream water during the course of the experimental addition is evaluated. Simulations were performed for 1990-2002. Experimental N addition began in 1991. INCA-N was able to successfully reproduce stream and soil water dynamics before and during the experiment. While INCA-N did not correctly simulate the lag between the start of N addition and NO 2 3 breakthrough, the model was able to simulate the state change resulting from increased N deposition. Sensitivity analysis showed that model behaviour was controlled primarily by parameters related to hydrology and vegetation dynamics and secondarily by in-soil processes.

Tracking larval insect movement within soil using high resolution X-ray microtomography

1. In contrast to above-ground insects, comparatively little is known about the behaviour of subterranean insects, due largely to the difficulty of studying them in situ. 2. The movement of newly hatched (neonate) clover root weevil (Sitona lepidus L. Coleoptera: Curculinidae) larvae was studied non-invasively using recently developed high resolution X-ray microtomography. 3. The movement and final position of S. lepidus larvae in the soil was reliably established using X-ray microtomography, when compared with larval positions that were determined by destructively sectioning the soil column. 4. Newly hatched S. lepidus larvae were seen to attack the root rhizobial nodules of their host plant, white clover (Trifolium repens L.). Sitona lepidus larvae travelled between 9 and 27 mm in 9 h at a mean speed of 1.8 mm h(-1). 5. Sitona lepidus larvae did not move through the soil in a linear manner, but changed trajectory in both the lateral and vertical planes.

Voices from the aid 'chain': The personal dynamics of care

This paper uses a simplified model of the aid 'chain' to explore some causes and consequences of breakdown in communication. Although the rhetoric of Northern-based donors is awash with words such as 'partnership' and 'inclusion' when dealing with their Southern-based partners, the situation in practice is different. Unequal power relationships sometimes result in donor imposition of Perspectives and values. It is our contention, based on a collective experience of fifty-four years in a Nigerian-based non-governmental development organization (NGDO), the Diocesan Development Services (DDS), that much of the driving force behind the successes and problems faced by the institution was founded on relationships that evolved between individuals. In order to understand why things happened the way they did it is necessary to begin with the human element that cannot be condensed into objects or categories. While injudicious donor interference bad damaging repercussions, our experience suggests that care and consideration flow throughout the aid chain and actions are not malevolent. Breakdowns can be attributed to a number of factors, with the over-riding one being pressures operating at the personal level that emanate from within the institution itself and the larger community. The paper analyses three experiences using institutional ethnography theory and methodologies as a basis. Examples taken address the influence key donor personnel had in the function of DDS, and how these changed with time. The mission, policies and even procedures of the donor did not change markedly over thirty-two years, but each changing desk officer had their own philosophy and approach and a different interpretation of their own institutional policies. Hence while the 'macro' has an influence it is mediated via individual interpretation. In our view, the importance of people-people relationships is particularly understated in development literature where emphasis gravitates towards the aggregate and global.

Phosphorus dynamics in the ditch system of a restored peat wetland

The restoration of wetlands as bird habitats often involves the maintenance of a fluctuating water regime by careful, localised ditch water management using pumps and sluices. However, there is evidence in the literature to Suggest that alternate flood/drainage cycles can accelerate nutrient cycling and transport within the soil and, therefore, pose a threat to water quality through the process of eutrophication. This study focused on the dynamics and losses of soil P in a recently re-wetted, eutrophic fen peat developed on alluvium ill South west England. During the 2-year Study (2001 and 2002), soil water tensiometry revealed that the field water table (fluctuating annually between +20 and 60 cm relative to ground level) was extensively influenced across most of the 8.4 ha field site by the management of the adjacent ditch water levels. This conservation-led, prescribed water balance was facilitated by the high hydraulic conductivity (1.1 x 10(-s) ms(-1)) of the lower (70-140 cm), degraded layer of peat. However, only during a 7-day period of water table drawdown by intermittent pump drainage, approximately 45 g ha(-1) of dissolved reactive P (DRP) entered the pumped ditch from the field via this degraded layer. Summer rainfall events >35 mm d(-1) also coincided with significant peaks ill ditch water P concentration (up to 200 mu g L-1 DRP). Even larger peaks (Up to 700 mu g L-1 DRP) Occurred With the annual onset of autumn reflooding. These episodic P loss events pose a serious potential threat to biological water quality. (C) 2009 Elsevier B.V. All rights reserved.

Assessing nitrogen dynamics in European ecosystems, integrating measurement and modelling: conclusions

This contribution closes this special issue of Hydrology and Earth System Sciences concerning the assessment of nitrogen dynamics in catchments across Europe within a semi-distributed Integrated Nitrogen model for multiple source assessment in Catchments (INCA). New developments in the understanding of the factors and processes determining the concentrations and loads of nitrogen are outlined. The ability of the INCA model to simulate the hydrological and nitrogen dynamics of different European ecosystems is assessed and the results of the first scenario analyses investigating the impacts of deposition, climatic and land-use change on the nitrogen dynamics are summarised. Consideration is given as to how well the model has performed as a generic too] for describing the nitrogen dynamics of European ecosystems across Arctic, Maritime. Continental and Mediterranean climates, its role in new research initiatives and future research requirements.

Nutrient monitoring, simulation and management within a major lowland UK river system: the Kennet

This paper describes an assessment of the nitrogen and phosphorus dynamics of the River Kennet in the south east of England. The Kennet catchment (1200 km(2)) is a predominantly groundwater fed river impacted by agricultural and sewage sources of nutrient (nitrogen and phosphorus) pollution. The results from a suite of simulation models are integrated to assess the key spatial and temporal variations in the nitrogen (N) and phosphorus (P) chemistry, and the influence of changes in phosphorous inputs from a Sewage Treatment Works on the macrophyte and epiphyte growth patterns. The models used are the Export Co-efficient model, the Integrated Nitrogen in Catchments model, and a new model of in-stream phosphorus and macrophyte dynamics: the 'Kennet' model. The paper concludes with a discussion on the present state of knowledge regarding the water quality functioning, future research needs regarding environmental modelling and the use of models as management tools for large, nutrient impacted riverine systems. (C) 2003 IMACS. Published by Elsevier B.V. All rights reserved.