Transcriptomic analysis of Enterohaemorrhagic Escherichia coli O157:H7 in response to plant extracts

Enterohaemorrhagic Escherichia coli (EHEC) are a group of food and contact-borne pathogens responsible for haemorrhagic colitis. The bacteria can be transmitted by contaminated meat, but importantly, also by plants. The bacteria can use plants as an alternative host, where they associate with both the leaves and the roots. Colonisation in the rhizosphere of plants is thought to be the main habitat for colonisation. Four different plant species, commonly associated with EHEC outbreaks, were infected with EHEC O157:H7 isolates Sakai and TUV 93-0 over ten days to assess the colonisation potential of the bacteria in both the phyllosphere and rhizosphere of plants. The rhizosphere was found to sustain a higher population level of bacteria over time in comparison to the phyllosphere, yet both strains were unable to utilize root exudates for growth. Global gene expression changes of EHEC O157:H7 strain Sakai were measured in response to plant extracts such as leaf lysates, root exudates and leaf cell wall polysaccharides from spinach cultivar Amazon and lettuce cultivar Salinas. Microarrays analysis showed a significant change in expression of 17 % of genes on exposure to leaf lysates of spinach. A more specific response was seen to spinach leaf cell wall polysaccharides with only a 1.5 % change. In contrast, when exposed to lettuce leaf cell wall polysaccharides a higher change of 4.8 % was seen. Genes that were differentially expressed belonged to multiple functional groups, including metabolism, indicating the utilization of plant-specific polysaccharides. Several areas of further investigation have been determined from this project, including the importance of culturing bacterial strains at a relevant temperature, the proposed lack of the type III secretion system in plant colonization by EHEC O157:H7 and the utilization of plant components for growth and persistence in the plant environment.

CDASH: a cloud-enabled program for structure solution from powder diffraction data

The simulated annealing approach to crystal structure determination from powder diffraction data, as implemented in the DASH program, is readily amenable to parallelization at the individual run level. Very large scale increases in speed of execution can be achieved by distributing individual DASH runs over a network of computers. The CDASH program delivers this by using scalable on-demand computing clusters built on the Amazon Elastic Compute Cloud service. By way of example, a 360 vCPU cluster returned the crystal structure of racemic ornidazole (Z0 = 3, 30 degrees of freedom) ca 40 times faster than a typical modern quad-core desktop CPU. Whilst used here specifically for DASH, this approach is of general applicability to other packages that are amenable to coarse-grained parallelism strategies.

Environmental controls on the distribution and diversity of lentic Chironomidae (Insecta: Diptera) across an altitudinal gradient in tropical South America

To predict the response of aquatic ecosystems to future global climate change, data on the ecology and distribution of keystone groups in freshwater ecosystems are needed. In contrast to mid- and high-latitude zones, such data are scarce across tropical South America (Neotropics). We present the distribution and diversity of chironomid species using surface sediments of 59 lakes from the Andes to the Amazon (0.1–17°S and 64–78°W) within the Neotropics. We assess the spatial variation in community assemblages and identify the key variables influencing the distributional patterns. The relationships between environmental variables (pH, conductivity, depth, and sediment organic content), climatic data, and chironomid assemblages were assessed using multivariate statistics (detrended correspondence analysis and canonical correspondence analysis). Climatic parameters (temperature and precipitation) were most significant in describing the variance in chironomid assemblages. Temperature and precipitation are both predicted to change under future climate change scenarios in the tropical Andes. Our findings suggest taxa of Orthocladiinae, which show a preference to cold high-elevation oligotrophic lakes, will likely see range contraction under future anthropogenic-induced climate change. Taxa abundant in areas of high precipitation, such as Micropsectra and Phaenopsectra, will likely become restricted to the inner tropical Andes, as the outer tropical Andes become drier. The sensitivity of chironomids to climate parameters makes them important bio-indicators of regional climate change in the Neotropics. Furthermore, the distribution of chironomid taxa presented here is a vital first step toward providing urgently needed autecological data for interpreting fossil chironomid records of past ecological and climate change from the tropical Andes.

Concordant Phylogeographies of 2 Malaria Vectors Attest to Common Spatial and Demographic Histories

The phylogeography of South American lineages is a topic of heated debate. Although a single process is unlikely to describe entire ecosystems, related species, which incur similar habitat limitations, can inform the history for a subsection of assemblages. We compared the phylogeographic patterns of the cytochrome oxidase I marker from Anopheles triannulatus (N = 72) and previous results for A. darlingi (N = 126) in a broad portion of their South American distributions. Both species share similar population subdivisions, with aggregations northeast of the Amazon River, in southern coastal Brazil and 2 regions in central Brazil. The average (ST) between these groups was 0.39 for A. triannulatus. Populations northeast of the Amazon and in southeastern Brazil are generally reciprocally monophyletic to the remaining groups. Based on these initial analyses, we constructed the a priori hypothesis that the Amazon and regions of high declivity pose geographic barriers to dispersal in these taxa. Mantel tests confirmed that these areas block gene flow for more than 1000 km for both species. The efficacy of these impediments was tested using landscape genetics, which could not reject our a priori hypothesis but did reject simpler scenarios. Results form summary statistics and phylogenetics suggest that both lineages originated in central Amazonia (south of the Amazon River) during the late Pleistocene (579 000 years ago) and that they followed the same paths of expansion into their contemporary distributions. These results may have implications for other species sharing similar ecological limitations but probably are not applicable as a general paradigm of Neotropical biogeography.

Spatial expansion and population structure of the neotropical malaria vector, Anopheles darlingi (Diptera: Culicidae)

Extensive population structuring is known to occur in Anopheles darlingi, the primary malaria vector of the Neotropics. We analysed the phylogeographic structure of the species using the mitochondrial cytochrome oxidase I marker. Diversity is divided into six main population groups in South America: Colombia, central Amazonia, southern Brazil, south-eastern Brazil, and two groups in north-east Brazil. The ancestral distribution of the taxon is hypothesized to be central Amazonia, and there is evidence of expansion from this region during the late Pleistocene. The expansion was not a homogeneous front, however, with at least four subgroups being formed due to geographic barriers. As the species spread, populations became isolated from each other by the Amazon River and the coastal mountain ranges of south-eastern Brazil and the Andes. Analyses incorporating distances around these barriers suggest that the entire South American range of An. darlingi is at mutation-dispersal-drift equilibrium. Because the species is distributed throughout such a broad area, the limited dispersal across some landscape types promotes differentiation between otherwise proximate populations. Moreover, samples from the An. darlingi holotype location in Rio de Janeiro State are substantially derived from all other populations, implying that there may be additional genetic differences of epidemiological relevance. The results obtained contribute to our understanding of gene flow in this species and allow the formulation of human mosquito health protocols in light of the potential population differences in vector capacity or tolerance to control strategies. (C) 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 97, 854-866.

Population-Based Multicentric Survey of Hepatitis B Infection and Risk Factor Differences among Three Regions in Brazil

This multicentric population-based study in Brazil is the first national effort to estimate the prevalence of hepatitis B (HBV) and risk factors in the capital cities of the Northeast. Central-West, and Federal Districts (2004-2005). Random multistage cluster sampling was used to select persons 13-69 years of age. Markers for HBV were tested by enzyme-linked immunosorbent assay. The HBV genotypes were determined by sequencing hepatitis B surface antigen (HBsAg). Multivariate analyses and simple catalytic model were performed. Overall. 7,881 persons were inculded < 70% were not vaccinated. Positivity for HBsAg was less than 1% among non-vaccinated persons and genotypes A, D, and F co-circulated. The incidence of infection increased with age with similar force of infection in all regions. Males and persons having initiated sexual activity were associated with HBV infection in the two settings: healthcare jobs and prior hospitalization were risk factors in the Federal District. Our survey classified these regions as areas with HBV endemicity and highlighted the risk factors differences among the settings.

Insight into Anopheles (Nyssorhynchus) (Diptera: Culicidae) Species from Brazil

Anopheles (Nyssorhynchus) benarrochi s.l., Anopheles (Nyssorhynchus) oswaldoi s.l., and Anopheles (Nyssorhynchus) konderi s.l. collected in Acrelandia, state of Acre, Brazil, were identified based on morphological characters of the male genitalia, fourth-instar larvae, and pupae. Morphological variation was observed in the male genitalia of these species in comparison with specimens from other localities in Brazil. DNA sequence from the nuclear ribosomal second internal transcribed spacer of individuals identified as An. benarrochi s.l. by using male genitalia characteristics showed that the various morphological forms are conspecific but are distinct from An. benarrochi B from Colombia. Anopheles konderi s.l. and An. oswaldoi s.l. both misidentified as An. oswaldoi s.s. (Peryassu) throughout Brazil, may actually comprise at least two undescribed species. Diagnostic morphological characteristics of the male genitalia are provided to distinguish Anopheles benarrochi s.l., Anopheles oswaldoi s.l., and Anopheles konderi s.l. from morphologically similar species. Incrimination of An. oswaldoi s.s. in malaria transmission in Brazil needs further investigation because other undescribed species from Acre may have been confounded with this taxon.

Natural Plasmodium infections in Brazilian wild monkeys: Reservoirs for human infections?

Four hundred and forty-eight samples of total blood from wild monkeys living in areas where human autochthonous malaria cases have been reported were screened for the presence of Plasmodium using microscopy and PCR analysis. Samples came from the following distinct ecological areas of Brazil: Atlantic forest (N = 140), semideciduous Atlantic forest (N = 257) and Cerrado (a savannah-like habitat) (N = 51). Thick and thin blood smears of each specimen were examined and Plasmodium infection was screened by multiplex polymerase chain reaction (multiplex PCR). The frequency of Plasmodium infections detected by PCR in Alouatta guariba clamitans in the Sao Paulo Atlantic forest was 11.3% or 8/71 (5.6% for Plasmodium malariae and 5.6% for Plasmodium vivax) and one specimen was positive for Plasmodium falciparum (1.4%); Callithrix sp. (N = 30) and Cebus apella (N = 39) specimens were negative by PCR tests. Microscopy analysis was negative for all specimens from the Atlantic forest. The positivity rate for Alouatta caraya from semideciduous Atlantic forest was 6.8% (16/235) in the PCR tests (5.5, 0.8 and 0.4% for P. malariae, P. falciparum and P. vivax, respectively), while C apella specimens were negative. Parasitological examination of I he samples using thick smears revealed Plasmodium sp. infections in only seven specimens, which had few parasites (3.0%). Monkeys from the Cerrado (a savannah-like habitat) (42 specimens of A. caraya, 5 of Callithrix jacchus and 4 of C. apella) were negative in both tests. The parasitological prevalence of P. vivax and P. malariae in wild monkeys from Atlantic forest and semideciduous Atlantic forest and the finding of a positive result for P.falciparum in Alouatta from both types of forest support the hypothesis that monkeys belonging to this genus could be a potential reservoir. Furthermore, these findings raise the question of the relationship between simian and autochthonous human malaria in extra-Amazonian regions. (C) 2008 Elsevier B.V. All rights reserved.

Mechanisms Associated with Large Daily Rainfall Events in Northeast Brazil

The mechanisms resulting in large daily rainfall events in Northeast Brazil are analyzed using data filtering to exclude periods longer than 30 days. Composites of circulation fields that include all independent events do not reveal any obvious forcing mechanisms as multiple patterns contribute to Northeast Brazil precipitation variability. To isolate coherent patterns, subsets of events are selected based on anomalies that precede the Northeast Brazil precipitation events at different locations. The results indicate that at 10 degrees S, 40 degrees W, the area of lowest annual rainfall in Brazil, precipitation occurs mainly in association with trailing midlatitude synoptic wave trains originating in either hemisphere. Closer to the equator at 5 degrees S, 37.5 degrees W, an additional convection precursor is found to the west, with a spatial structure consistent with that of a Kelvin wave. Although these two sites are located within only several hundred kilometers of each other and the midlatitude patterns that induce precipitation appear to be quite similar, the dates on which large precipitation anomalies occur at each location are almost entirely independent, pointing to separate forcing mechanisms.

Summertime moisture transport over Southeastern South America and extratropical cyclones behavior during inter-El Nio events

The impact of the inter-El Nio (EN) variability on the moisture availability over Southeastern South America (SESA) is investigated. Also, an automatic tracking scheme was used to analyze the extratropical cyclones properties (system density - SD and central pressure - CP) in this region. During the austral summer period from 1977-2000, the differences for the upper-level wave train anomaly composites seem to determine the rainfall composite differences. In fact, the positive rainfall anomalies over most of the SESA domain during the strong EN events are explained by an upper-level cyclonic center over the tropics and an anticyclonic center over the eastern subtropical area. This pattern seems to contribute to upward vertical motion at 500 hPa and reinforcement of the meridional moisture transport from the equatorial Atlantic Ocean and western Amazon basin to the SESA region. These features may contribute to the positive SD and negative CP anomalies explaining part of the positive rainfall anomalies found there. On the other hand, negative rainfall anomalies are located in the northern part of SESA for the weak EN years when compared to those for the strong events. Also, positive anomalies are found in the southern part, albeit less intense. It was associated with the weakening of the meridional moisture transport from the tropics to the SESA that seems have to contributed with smaller SD and CP anomalies over the most part of subtropics, when compared to the strong EN years.

IPCC global coupled model simulations of the South America monsoon system

This study examines the variability of the South America monsoon system (SAMS) over tropical South America (SA). The onset, end, and total rainfall during the summer monsoon are investigated using precipitation pentad estimates from the global precipitation climatology project (GPCP) 1979-2006. Likewise, the variability of SAMS characteristics is examined in ten Intergovernmental Panel on Climate Change (IPCC) global coupled climate models in the twentieth century (1981-2000) and in a future scenario of global change (A1B) (2081-2100). It is shown that most IPCC models misrepresent the intertropical convergence zone and therefore do not capture the actual annual cycle of precipitation over the Amazon and northwest SA. Most models can correctly represent the spatiotemporal variability of the annual cycle of precipitation in central and eastern Brazil such as the correct phase of dry and wet seasons, onset dates, duration of rainy season and total accumulated precipitation during the summer monsoon for the twentieth century runs. Nevertheless, poor representation of the total monsoonal precipitation over the Amazon and northeast Brazil is observed in a large majority of the models. Overall, MI-ROC3.2-hires, MIROC3.2-medres and MRI-CGCM3.2.3 show the most realistic representation of SAMS`s characteristics such as onset, duration, total monsoonal precipitation, and its interannual variability. On the other hand, ECHAM5, GFDL-CM2.0 and GFDL-CM2.1 have the least realistic representation of the same characteristics. For the A1B scenario the most coherent feature observed in the IPCC models is a reduction in precipitation over central-eastern Brazil during the summer monsoon, comparatively with the present climate. The IPCC models do not indicate statistically significant changes in SAMS onset and demise dates for the same scenario.

The land-atmosphere water flux in the tropics

Tropical vegetation is a major source of global land surface evapotranspiration, and can thus play a major role in global hydrological cycles and global atmospheric circulation. Accurate prediction of tropical evapotranspiration is critical to our understanding of these processes under changing climate. We examined the controls on evapotranspiration in tropical vegetation at 21 pan-tropical eddy covariance sites, conducted a comprehensive and systematic evaluation of 13 evapotranspiration models at these sites, and assessed the ability to scale up model estimates of evapotranspiration for the test region of Amazonia. Net radiation was the strongest determinant of evapotranspiration (mean evaporative fraction was 0.72) and explained 87% of the variance in monthly evapotranspiration across the sites. Vapor pressure deficit was the strongest residual predictor (14%), followed by normalized difference vegetation index (9%), precipitation (6%) and wind speed (4%). The radiation-based evapotranspiration models performed best overall for three reasons: (1) the vegetation was largely decoupled from atmospheric turbulent transfer (calculated from X decoupling factor), especially at the wetter sites; (2) the resistance-based models were hindered by difficulty in consistently characterizing canopy (and stomatal) resistance in the highly diverse vegetation; (3) the temperature-based models inadequately captured the variability in tropical evapotranspiration. We evaluated the potential to predict regional evapotranspiration for one test region: Amazonia. We estimated an Amazonia-wide evapotranspiration of 1370 mm yr(-1), but this value is dependent on assumptions about energy balance closure for the tropical eddy covariance sites; a lower value (1096 mm yr(-1)) is considered in discussion on the use of flux data to validate and interpolate models.

An improved estimate of leaf area index based on the histogram analysis of hemispherical photographs

Leaf area index (LAI) is a key parameter that affects the surface fluxes of energy, mass, and momentum over vegetated lands, but observational measurements are scarce, especially in remote areas with complex canopy structure. In this paper we present an indirect method to calculate the LAI based on the analyses of histograms of hemispherical photographs. The optimal threshold value (OTV), the gray-level required to separate the background (sky) and the foreground (leaves), was analytically calculated using the entropy crossover method (Sahoo, P.K., Slaaf, D.W., Albert, T.A., 1997. Threshold selection using a minimal histogram entropy difference. Optical Engineering 36(7) 1976-1981). The OTV was used to calculate the LAI using the well-known gap fraction method. This methodology was tested in two different ecosystems, including Amazon forest and pasturelands in Brazil. In general, the error between observed and calculated LAI was similar to 6%. The methodology presented is suitable for the calculation of LAI since it is responsive to sky conditions, automatic, easy to implement, faster than commercially available software, and requires less data storage. (C) 2008 Elsevier B.V. All rights reserved.

Cloud condensation nuclei from biomass burning during the Amazonian dry-to-wet transition season

Aircraft measurements of cloud condensation nuclei (CCN) during the Large-Scale Biosphere-Atmosphere Experiment in Amazonia (LBA) were conducted over the Southwestern Amazon region in September-October 2002, to emphasize the dry-to-wet transition season. The CCN concentrations were measured for values within the range 0.1-1.0% of supersaturation. The CCN concentration inside the boundary layer revealed a general decreasing trend during the transition from the end of the dry season to the onset of the wet season. Clean and polluted areas showed large differences. The differences were not so strong at high levels in the troposphere and there was evidence supporting the semi-direct aerosol effect in suppressing convection through the evaporation of clouds by aerosol absorption. The measurements also showed a diurnal cycle following biomass burning activity. Although biomass burning was the most important source of CCN, it was seen as a source of relatively efficient CCN, since the increase was significant only at high supersaturations.

Precipitation diurnal cycle and summer climatology assessment over South America: An evaluation of Regional Climate Model version 3 simulations

Regional Climate Model version 3 (RegCM3) simulations of 17 summers (1988-2004) over part of South America south of 5 degrees S were evaluated to identify model systematic errors. Model results were compared to different rainfall data sets (Climate Research Unit (CRU), Climate Prediction Center (CPC), Global Precipitation Climatology Project (GPCP), and National Centers for Environmental Prediction (NCEP) reanalysis), including the five summers mean (1998-2002) precipitation diurnal cycle observed by the Tropical Rainfall Measuring Mission (TRMM)-Precipitation Radar (PR). In spite of regional differences, the RegCM3 simulates the main observed aspects of summer climatology associated with the precipitation (northwest-southeast band of South Atlantic Convergence Zone (SACZ)) and air temperature (warmer air in the central part of the continent and colder in eastern Brazil and the Andes Mountains). At a regional scale, the main RegCM3 failures are the underestimation of the precipitation in the northern branch of the SACZ and some unrealistic intense precipitation around the Andes Mountains. However, the RegCM3 seasonal precipitation is closer to the fine-scale analyses (CPC, CRU, and TRMM-PR) than is the NCEP reanalysis, which presents an incorrect north-south orientation of SACZ and an overestimation of its intensity. The precipitation diurnal cycle observed by TRMM-PR shows pronounced contrasts between Tropics and Extratropics and land and ocean, where most of these features are simulated by RegCM3. The major similarities between the simulation and observation, especially the diurnal cycle phase, are found over the continental tropical and subtropical SACZ regions, which present afternoon maximum (1500-1800 UTC) and morning minimum (0900-1200 UTC). More specifically, over the core of SACZ, the phase and amplitude of the simulated precipitation diurnal cycle are very close to the TRMM-PR observations. Although there are amplitude differences, the RegCM3 simulates the observed nighttime rainfall in the eastern Andes Mountains, over the Atlantic Ocean, and also over northern Argentina. The main simulation deficiencies are found in the Atlantic Ocean and near the Andes Mountains. Over the Atlantic Ocean the convective scheme is not triggered; thus the rainfall arises from the grid-scale scheme and therefore differs from the TRMM-PR. Near the Andes, intense (nighttime and daytime) simulated precipitation could be a response of an incorrect circulation and topographic uplift. Finally, it is important to note that unlike most reported bias of global models, RegCM3 does not trigger the moist convection just after sunrise over the southern part of the Amazon.