Genomics of pyrrolnitrin biosynthetic loci: evidence for conservation and whole-operon mobility within Gram-negative bacteria

Pyrrolnitrin (PRN) is a tryptophan-derived secondary metabolite produced by a narrow range of Gram-negative bacteria. The PRN biosynthesis by rhizobacteria presumably has a key role in their life strategies and in the biocontrol of plant diseases. The biosynthetic operon that encodes the pathway that converts tryptophan to PRN is composed of four genes, prnA through D, whose diversity, genomic context and spread over bacterial genomes are poorly understood. Therefore, we launched an endeavour aimed at retrieving, by in vitro and in silico means, diverse bacteria carrying the prnABCD biosynthetic loci in their genomes. Analysis of polymorphisms of the prnD gene sequences revealed a high level of conservation between Burkholderia, Pseudomonas and Serratia spp. derived sequences. Whole-operon- and prnD-based phylogeny resulted in tree topologies that are incongruent with the taxonomic status of the evaluated strains as predicted by 16S rRNA gene phylogeny. The genomic composition of c. 20 kb DNA fragments containg the PRN operon varied in different strains. Highly conserved and distinct transposase-encoding genes surrounding the PRN biosynthetic operons of Burkholderia pseudomallei strains were found. A prnABCD-deprived genomic region in B. pseudomallei strain K96243 contained the same gene composition as, and shared high homology with, the flanking regions of the PRN operon in B. pseudomallei strains 668, 1106a and 1710b. Our results strongly suggest that the PRN biosynthetic operon is mobile. The extent, frequency and promiscuity of this mobility remain to be understood.

Bacterial communities and biogeochemical transformations of iron and sulfur in a high saltmarsh soil profile

Microbial community structure in saltmarsh soils is stratified by depth and availability of electron acceptors for respiration. However, the majority of the microbial species that are involved in the biogeochemical transformations of iron (Fe) and sulfur (S) in such environments are not known. Here we examined the structure of bacterial communities in a high saltmarsh soil profile and discuss their potential relationship with the geochemistry of Fe and S. Our data showed that the soil horizons Ag (oxic-suboxic), Bg (suboxic), Cri (anoxic with low concentration of pyrite Fe) and Cr-2 (anoxic with high concentrations of pyrite Fe) have distinct geochemical and microbiological characteristics. In general, total S concentration increased with depth and was correlated with the presence of pyrite Fe. Soluble + exchangable-Fe, pyrite Fe and acid volatile sulfide Fe concentrations also increased with depth, whereas ascorbate extractable-Fe concentrations decreased. The occurrence of reduced forms of Fe in the horizon Ag and oxidized Fe in horizon Cr-2 suggests that the typical redox zonation, common to several marine sediments, does not occur in the saltmarsh soil profile studied. Overall, the bacterial community structure in the horizon Ag and Cr-2 shared low levels of similarity, as compared to their adjacent horizons, Bg and Cr-1, respectively. The phylogenetic analyses of bacterial 16S rRNA gene sequences from clone libraries showed that the predominant phylotypes in horizon Ag were related to Alphaproteobacteria and Bacteroidetes. In contrast, the most abundant phylotypes in horizon Cr-2 were related to Deltaproteo-bacteria, Chloroflexi, Deferribacteres and Nitrospira. The high frequency of sequences with low levels of similarity to known bacterial species in horizons Ag and Cr-2 indicates that the bacterial communities in both horizons are dominated by novel bacterial species. (c) 2008 Elsevier Ltd. All rights reserved.

Untangling associations between chironomid taxa in Neotropical streams using local and landscape filters

1. Analyses of species association have major implications for selecting indicators for freshwater biomonitoring and conservation, because they allow for the elimination of redundant information and focus on taxa that can be easily handled and identified. These analyses are particularly relevant in the debate about using speciose groups (such as the Chironomidae) as indicators in the tropics, because they require difficult and time-consuming analysis, and their responses to environmental gradients, including anthropogenic stressors, are poorly known. 2. Our objective was to show whether chironomid assemblages in Neotropical streams include clear associations of taxa and, if so, how well these associations could be explained by a set of models containing information from different spatial scales. For this, we formulated a priori models that allowed for the influence of local, landscape and spatial factors on chironomid taxon associations (CTA). These models represented biological hypotheses capable of explaining associations between chironomid taxa. For instance, CTA could be best explained by local variables (e.g. pH, conductivity and water temperature) or by processes acting at wider landscape scales (e.g. percentage of forest cover). 3. Biological data were taken from 61 streams in Southeastern Brazil, 47 of which were in well-preserved regions, and 14 of which drained areas severely affected by anthropogenic activities. We adopted a model selection procedure using Akaike`s information criterion to determine the most parsimonious models for explaining CTA. 4. Applying Kendall`s coefficient of concordance, seven genera (Tanytarsus/Caladomyia, Ablabesmyia, Parametriocnemus, Pentaneura, Nanocladius, Polypedilum and Rheotanytarsus) were identified as associated taxa. The best-supported model explained 42.6% of the total variance in the abundance of associated taxa. This model combined local and landscape environmental filters and spatial variables (which were derived from eigenfunction analysis). However, the model with local filters and spatial variables also had a good chance of being selected as the best model. 5. Standardised partial regression coefficients of local and landscape filters, including spatial variables, derived from model averaging allowed an estimation of which variables were best correlated with the abundance of associated taxa. In general, the abundance of the associated genera tended to be lower in streams characterised by a high percentage of forest cover (landscape scale), lower proportion of muddy substrata and high values of pH and conductivity (local scale). 6. Overall, our main result adds to the increasing number of studies that have indicated the importance of local and landscape variables, as well as the spatial relationships among sampling sites, for explaining aquatic insect community patterns in streams. Furthermore, our findings open new possibilities for the elimination of redundant data in the assessment of anthropogenic impacts on tropical streams.

Non-random reef use by fishes at two dominant zones in a tropical, algal-dominated coastal reef

Habitat use and the processes which determine fish distribution were evaluated at the reef flat and reef crest zones of a tropical, algal-dominated reef. Our comparisons indicated significant differences in the majority of the evaluated environmental characteristics between zones. Also, significant differences in the abundances of twelve, from thirteen analyzed species, were observed within and between-sites. According to null models, non-random patterns of species co-occurrences were significant, suggesting that fish guilds in both zones were non-randomly structured. Unexpectedly, structural complexity negatively affected overall species richness, but had a major positive influence on highly site-attached species such as a damselfish. Depth and substrate composition, particularly macroalgae cover, were positive determinants for the fish assemblage structure in the studied reef, prevailing over factors such as structural complexity and live coral cover. Our results are conflicting with other studies carried out in coral-dominated reefs of the Caribbean and Pacific, therefore supporting the idea that the factors which may potentially influence reef fish composition are highly site-dependent and variable.

SZNAJD MODEL AND PROPORTIONAL ELECTIONS: THE ROLE OF THE TOPOLOGY OF THE NETWORK

The Sznajd model (SM) has been employed with success in the last years to describe opinion propagation in a community. In particular, it has been claimed that its transient is able to reproduce some scale properties observed in data of proportional elections, in different countries, if the community structure (the network) is scale-free. In this work, we investigate the properties of the transient of a particular version of the SM, introduced by Bernardes and co-authors in 2002. We studied the behavior of the model in networks of different topologies through the time evolution of an order parameter known as interface density, and concluded that regular lattices with high dimensionality also leads to a power-law distribution of the number of candidates with v votes. Also, we show that the particular absorbing state achieved in the stationary state (or else, the winner candidate), is related to a particular feature of the model, that may not be realistic in all situations.

MODELING THE EVOLUTION OF COMPLEX NETWORKS THROUGH THE PATH-STAR TRANSFORMATION AND OPTIMAL MULTIVARIATE METHODS

The topology of real-world complex networks, such as in transportation and communication, is always changing with time. Such changes can arise not only as a natural consequence of their growth, but also due to major modi. cations in their intrinsic organization. For instance, the network of transportation routes between cities and towns ( hence locations) of a given country undergo a major change with the progressive implementation of commercial air transportation. While the locations could be originally interconnected through highways ( paths, giving rise to geographical networks), transportation between those sites progressively shifted or was complemented by air transportation, with scale free characteristics. In the present work we introduce the path-star transformation ( in its uniform and preferential versions) as a means to model such network transformations where paths give rise to stars of connectivity. It is also shown, through optimal multivariate statistical methods (i.e. canonical projections and maximum likelihood classification) that while the US highways network adheres closely to a geographical network model, its path-star transformation yields a network whose topological properties closely resembles those of the respective airport transportation network.

Ecosystem health of Montevideo coastal zone: A multi approach using some different benthic indicators to improve a ten-year-ago assessment

A study on the benthic ecosystem health was performed to assess the environmental quality of Montevideo coastal zone, in view of the construction of a new sanitation system. Data were compared to previous research undertaken 10 years ago, and biochemical composition of organic matter, heavy metals, organic matter, phytopigments, benthic diatoms, macrofauna community structure and a biotic index (AMBI) were used as proxies. Results indicate an environmental quality-gradient, with the worst conditions within the inner stations of Montevideo Bay and an improvement towards the adjacent coastal zone. Higher levels of chromium, lead, phaeopigments, organic biopolymers and poor benthic macrofauna and diatom communities, characterised the hypertrophic innermost portion of Montevideo Bay. Data indicated a clear deterioration of the adjacent coastal zone comparatively to that observed 10 years ago. The complementary use of approaches not applied before (benthic diatoms and organic biopolymers) with those formerly applied improve our assessment of the trophic status and the environmental health of the area. (C) 2010 Elsevier B.V. All rights reserved.

The Staircase Structure of the Southern Brazilian Continental Shelf

We show some evidences that the Southeastern Brazilian Continental Shelf (SBCS) has a devil's staircase structure, with a sequence of scarps and terraces with widths that obey fractal formation rules. Since the formation of these features is linked with the sea-level variations, we say that the sea level changes in an organized pulsating way. Although the proposed approach was applied in a particular region of the Earth, it is suitable to be applied in an integrated way to other shelves around the world, since the analyses favor the revelation of the global sea-level variations. Copyright (C) 2009 M. S. Baptista and L. A. Conti.

Foraging by the omnivorous crab Pachygrapsus transversus affects the structure of assemblages on sub-tropical rocky shores

The roles of herbivory and predation in determining the structure and diversity of communities have been tested across most intertidal systems. In contrast, the importance of omnivorous consumers remains untested in many rocky shore communities. We tested the role of a small omnivorous crab in an intertidal food web on rocky shores of the sub-tropical southwest Atlantic. Exclusion of the grapsid crab Pachygrapsus transversus in the field resulted in significant changes in the abundance of functional groups in the sublittoral fringe of sheltered shores, where the dominant cover changed from a suite of macroalgae to an assemblage of filter-feeding animals (ascidians, sponges, mussels). In contrast, limpets, whelks, large crabs and fish did not significantly affect community composition of the assemblage. To examine the omnivorous feeding pattern of P. transversus, we did laboratory experiments to test its foraging behaviour among animal and algal groups. The crab showed selective behaviour, preferring invertebrate groups to macroalgae, and opportunistic behaviour among types of prey within those major groups. According to our results, the role of slow-moving and large fast-moving consumers is apparently negligible compared to the effect of an omnivorous consumer. P. transversus plays an important role in determining the intertidal community composition on these subtropical rocky shores, causing changes in the balance of functional groups and controlling invasive species.

Influence of the bacterioplankton community of a tropical eutrophic lagoon on the bacterial community of its neighbouring ocean

The Rodrigo de Freitas lagoon (RFL) is a tropical eutrophic coastal ecosystem located in the urban area of Rio de Janeiro, Brazil. This environment consists of freshwater but has communication with the ocean through a channel (Jardim de Alah`s Channel). The aim of this study was to evaluate the influence of lagoon water on the nearby ocean using molecular and traditional microbiological methods. We hypothesised that due to the eutrophic low-salinity environment, the bacterioplankton community from the RFL would have a native ""brackish"" composition influenced by both freshwater and marine phylotypes, and that bacterial phylotypes of this community would be detected in oceanic samples closer to the channel between the lagoon and the ocean. The cultivation and microscopy experiments clearly showed this influence. Bacterial cell counts revealed that the greater amounts of bacterial cells present in the lagoon increased the observed values seen at oceanic stations near the channel. The Denaturing gradient gel eletrophoresis community profiles also showed a clear influence of Rodrigo de Freitas lagoon waters on the adjacent beaches. The band patterns found for the stations near the channel showed that these communities were mixtures of the communities of the lagoon and sea, and as the distance from the channel increased, the samples became more similar to ocean bacterial communities. A 16S rRNA gene clone library was constructed using a sample acquired from the connection point between the lagoon and the ocean. Around 52% of the sequences in the library showed similarity to the genus Proteobacteria (1% Alpha, 21% Beta, 19% Gamma and 29% unclassified Proteobacteria), and the second most abundant genus was Bacteroidetes, with 15% of the total clones. The results showed that the structure of the bacterial community had both freshwater and marine characteristics.

Bacterial soil community in a Brazilian sugarcane field

The assessment of bacterial communities in soil gives insight into microbial behavior under prevailing environmental conditions. In this context, we assessed the composition of soil bacterial communities in a Brazilian sugarcane experimental field. The experimental design encompassed plots containing common sugarcane (variety SP80-1842) and its transgenic form (IMI-1 - imazapyr herbicide resistant). Plants were grown in such field plots in a completely randomized design with three treatments, which addressed the factors transgene and imazapyr herbicide application. Soil samples were taken at three developmental stages during plant growth and analyzed using 16S ribosomal RNA (rRNA)-based PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and clone libraries. PCR-DGGE fingerprints obtained for the total bacterial community and specific bacterial groups - Actinobacteria, Alphaproteobacteria and Betaproteobacteria - revealed that the structure of these assemblages did not differ over time and among treatments. Nevertheless, slight differences among 16S rRNA gene clone libraries constructed from each treatment could be observed at particular cut-off levels. Altogether, the libraries encompassed a total of eleven bacterial phyla and the candidate divisions TM7 and OP10. Clone sequences affiliated with the Proteobacteria, Actinobacteria, Firmicutes and Acidobacteria were, in this order, most abundant. Accurate phylogenetic analyses were performed for the phyla Acidobacteria and Verrucomicrobia, revealing the structures of these groups, which are still poorly understood as to their importance for soil functioning and sustainability under agricultural practices.

Bacterial community in the rhizosphere and rhizoplane of wild type and transgenic eucalyptus

The rhizosphere is a niche exploited by a wide variety of bacteria. The expression of heterologous genes by plants might become a factor affecting the structure of bacterial communities in the rhizosphere. In a greenhouse experiment, the bacterial community associated to transgenic eucalyptus, carrying the Lhcb1-2 genes from pea (responsible for a higher photosynthetic capacity), was evaluated. The culturable bacterial community associated to transgenic and wild type plants were not different in density, and the Amplified Ribosomal DNA Restriction Analysis (ARDRA) typing of 124 strains revealed dominant ribotypes representing the bacterial orders Burkholderiales, Rhizobiales, and Actinomycetales, the families Xanthomonadaceae, and Bacillaceae, and the genus Mycobacterium. Principal Component Analysis based on the fingerprints obtained by culture-independent Denaturing Gradient Gel Electrophoresis analysis revealed that Alphaproteobacteria, Betaproteobacteria and Actinobacteria communities responded differently to plant genotypes. Similar effects for the cultivation of transgenic eucalyptus to those observed when two genotype-distinct wild type plants are compared.

Evaluation of the aquatic macroinvertebrate community as a tool for monitoring a reservoir in the Pitangui river basin, Parana, Brazil

Evaluation of the aquatic macroinvertebrate community as a tool for monitoring a reservoir in the Pitangui river basin, Parana, Brazil. Benthic and nektonic macroinvertebrates play an important role in the structure and function of aquatic ecosystems and their distribution is influenced by chemical features of the substrate, vegetation composition, and water depth. Knowledge on the fauna contributes to the assessment of water quality and development of biodiversity conservation activities. Different biotic factors affecting the invertebrate community were evaluated in the Alagados reservoir, the main water source of the city of Ponta Grossa, Parana. In five different sampling points, 18,473 specimens of aquatic or semiaquatic macroinvertebrates were collected, belonging to 46 taxa of the phylla Annelida (Hirudinea and Oligochaeta), Mollusca (Gastropoda), Platyhelminthes (Turbellaria), Nematoda and Arthropoda (Arachnida, Crustacea and Insecta). This community was composed mainly of predators (45.7% of the taxa sampled), collectors and/or filterers (23.9%), scrapers (15.2%), shredders (13.0%) and detritivores (2.2%). Diversity (H`) and evenness (J) indices were significantly low for the sites examined, and H` ranged between 0.3301 and 1.0396. Regarding tolerance of organisms to organic pollution, more sensitive taxa were very rare (Plecoptera) or unusual (Trichoptera and Ephemeroptera). Among the more resistant groups are Chironomidae and Hirudinea, both fairly common in the samples. This study corroborates the importance of bioindicators as a tool to assess water quality for human consumption and for the conservation of aquatic environments, integrating physical, chemical and biological factors in monitoring programs.

Comparing species and measures of landscape structure as indicators of conservation importance

P>1. The use of indicators to identify areas of conservation importance has been challenged on several grounds, but nonetheless retains appeal as no more parsimonious approach exists. Among the many variants, two indicator strategies stand out: the use of indicator species and the use of metrics of landscape structure. While the first has been thoroughly studied, the same cannot be said about the latter. We aimed to contrast the relative efficacy of species-based and landscape-based indicators by: (i) comparing their ability to reflect changes in community integrity at regional and landscape spatial scales, (ii) assessing their sensitivity to changes in data resolution, and (iii) quantifying the degree to which indicators that are generated in one landscape or at one spatial scale can be transferred to additional landscapes or scales. 2. We used data from more than 7000 bird captures in 65 sites from six 10 000-ha landscapes with different proportions of forest cover in the Atlantic Forest of Brazil. Indicator species and landscape-based indicators were tested in terms of how effective they were in reflecting changes in community integrity, defined as deviations in bird community composition from control areas. 3. At the regional scale, indicator species provided more robust depictions of community integrity than landscape-based indicators. At the landscape scale, however, landscape-based indicators performed more effectively, more consistently and were also more transferable among landscapes. The effectiveness of high resolution landscape-based indicators was reduced by just 12% when these were used to explain patterns of community integrity in independent data sets. By contrast, the effectiveness of species-based indicators was reduced by 33%. 4. Synthesis and applications. The use of indicator species proved to be effective; however their results were variable and sensitive to changes in scale and resolution, and their application requires extensive and time-consuming field work. Landscape-based indicators were not only effective but were also much less context-dependent. The use of landscape-based indicators may allow the rapid identification of priority areas for conservation and restoration, and indicate which restoration strategies should be pursued, using remotely sensed imagery. We suggest that landscape-based indicators might often be a better, simpler, and cheaper strategy for informing decisions in conservation.

The modularity of seed dispersal: differences in structure and robustness between bat- and bird-fruit networks

In networks of plant-animal mutualisms, different animal groups interact preferentially with different plants, thus forming distinct modules responsible for different parts of the service. However, what we currently know about seed dispersal networks is based only on birds. Therefore, we wished to fill this gap by studying bat-fruit networks and testing how they differ from bird-fruit networks. As dietary overlap of Neotropical bats and birds is low, they should form distinct mutualistic modules within local networks. Furthermore, since frugivory evolved only once among Neotropical bats, but several times independently among Neotropical birds, greater dietary overlap is expected among bats, and thus connectance and nestedness should be higher in bat-fruit networks. If bat-fruit networks have higher nestedness and connectance, they should be more robust to extinctions. We analyzed 1 mixed network of both bats and birds and 20 networks that consisted exclusively of either bats (11) or birds (9). As expected, the structure of the mixed network was both modular (M = 0.45) and nested (NODF = 0.31); one module contained only birds and two only bats. In 20 datasets with only one disperser group, bat-fruit networks (NODF = 0.53 +/- A 0.09, C = 0.30 +/- A 0.11) were more nested and had a higher connectance than bird-fruit networks (NODF = 0.42 +/- A 0.07, C = 0.22 +/- A 0.09). Unexpectedly, robustness to extinction of animal species was higher in bird-fruit networks (R = 0.60 +/- A 0.13) than in bat-fruit networks (R = 0.54 +/- A 0.09), and differences were explained mainly by species richness. These findings suggest that a modular structure also occurs in seed dispersal networks, similar to pollination networks. The higher nestedness and connectance observed in bat-fruit networks compared with bird-fruit networks may be explained by the monophyletic evolution of frugivory in Neotropical bats, among which the diets of specialists seem to have evolved from the pool of fruits consumed by generalists.