Transfer origins in the conjugative Enterococcus faecalis plasmids pAD1 and pAM373: identification of the pAD1 nic site, a specific relaxase and a possible TraG-like protein

The Enterococcus faecalis conjugative plasmids pAD1 and pAM373 encode a mating response to the peptide sex pheromones cAD1 and cAM373 respectively. Sequence determination of both plasmids has recently been completed with strong similarity evident over many of the structural genes related to conjugation. pAD1 has two origins of transfer, with oriT1 being located within the repA determinant, whereas the more efficiently utilized oriT2 is located between orf53 and orf57 , two genes found in the present study to be essential for conjugation. We have found a similarly located oriT to be present in pAM373. oriT2 corresponds to about 285 bp based on its ability to facilitate mobilization by pAD1 when ligated to the shuttle vector pAM401; however, it was not mobilized by pAM373. In contrast, a similarly ligated fragment containing the oriT of pAM373 did not facilitate mobilization by pAD1 but was efficiently mobilized by pAM373. The oriT sites of the two plasmids each contained a homologous large inverted repeat (spanning about 140 bp) adjacent to a series of non-homologous short (6 bp) direct repeats. A hybrid construction containing the inverted repeat of pAM373 and direct repeats of pAD1 was mobilized efficiently by pAD1 but not by pAM373, indicating a significantly greater degree of specificity is associated with the direct repeats. Mutational (deletion) analyses of the pAD1 oriT2 inverted repeat structure suggested its importance in facilitating transfer or perhaps ligation of the ends of the newly transferred DNA strand. Analyses showed that Orf57 (to be called TraX) is the relaxase, which was found to induce a specific nick in the large inverted repeat inside oriT ; the protein also facilitated site-specific recombination between two oriT2 sites. Orf53 (to be called TraW) exhibits certain structural similarities to TraG-like proteins, although there is little overall homology.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72536/1/j.1365-2958.2002.03007.x.pd

Soil and water bioengineering: practice and research needs for reconciling natural hazard control and ecological restoration

Soil and water bioengineering is a technology that encourages scientists and practitioners to combine their knowledge and skills in the management of ecosystems with a common goal to maximize benefits to both man and the natural environment. It involves techniques that use plants as living building materials, for: (i) natural hazard control (e.g., soil erosion, torrential floods and landslides) and (ii) ecological restoration or nature-based re-introduction of species on degraded lands, river embankments, and disturbed environments. For a bioengineering project to be successful, engineers are required to highlight all the potential benefits and ecosystem services by documenting the technical, ecological, economic and social values. The novel approaches used by bioengineers raise questions for researchers and necessitate innovation from practitioners to design bioengineering concepts and techniques. Our objective in this paper, therefore, is to highlight the practice and research needs in soil and water bioengineering for reconciling natural hazard control and ecological restoration. Firstly, we review the definition and development of bioengineering technology, while stressing issues concerning the design, implementation, and monitoring of bioengineering actions. Secondly, we highlight the need to reconcile natural hazard control and ecological restoration by posing novel practice and research questions

Contrasting influences of inundation and land use on the rate of floodplain restoration

This study examined the assisted natural restoration of native vegetation in an Australian floodplain wetland where flows were reinstated and the river was reconnected to the floodplain, following cessation of agricultural cultivation. Extant vegetation was surveyed three times during an inundation event at plots with different land‐use histories. Restoration rate was more influenced by past land use than long‐term inundation frequency and success decreased with antecedent land‐use intensity. Prolonged land‐use history (>3 years cultivation) restricted restoration success. Sites with longer cultivation histories tended to have fewer aquatic species, more terrestrial species and exotic species. For example, amphibious responders with floating leaves were found only in reference plots and less frequently in farmed treatment plots. In this scenario, increased persistence of exotics and dryland species suggested alternative trajectories. Fields with a short land‐use history (1–3 years of clearing and cultivation) resembled undisturbed floodplain communities, consistent with a ‘field of dreams’ hypothesis. Although river–floodplain reconnections can restore wetlands, legacy effects of past land use may limit the pace and outcomes of restoration.Australian Postgraduate AwardAustralian Research Council. Grant Number: DE120102221ARC Centre of Excellence for Environmental Decisions Australian Research Council Linkage Project. Grant Number: LP088416

Dispelling urban myths about default uncertainty factors in chemical risk assessment - Sufficient protection against mixture effects?

© 2013 Martin et al.; licensee BioMed Central LtdThis article has been made available through the Brunel Open Access Publishing Fund.Assessing the detrimental health effects of chemicals requires the extrapolation of experimental data in animals to human populations. This is achieved by applying a default uncertainty factor of 100 to doses not found to be associated with observable effects in laboratory animals. It is commonly assumed that the toxicokinetic and toxicodynamic sub-components of this default uncertainty factor represent worst-case scenarios and that the multiplication of those components yields conservative estimates of safe levels for humans. It is sometimes claimed that this conservatism also offers adequate protection from mixture effects. By analysing the evolution of uncertainty factors from a historical perspective, we expose that the default factor and its sub-components are intended to represent adequate rather than worst-case scenarios. The intention of using assessment factors for mixture effects was abandoned thirty years ago. It is also often ignored that the conservatism (or otherwise) of uncertainty factors can only be considered in relation to a defined level of protection. A protection equivalent to an effect magnitude of 0.001-0.0001% over background incidence is generally considered acceptable. However, it is impossible to say whether this level of protection is in fact realised with the tolerable doses that are derived by employing uncertainty factors. Accordingly, it is difficult to assess whether uncertainty factors overestimate or underestimate the sensitivity differences in human populations. It is also often not appreciated that the outcome of probabilistic approaches to the multiplication of sub-factors is dependent on the choice of probability distributions. Therefore, the idea that default uncertainty factors are overly conservative worst-case scenarios which can account both for the lack of statistical power in animal experiments and protect against potential mixture effects is ill-founded. We contend that precautionary regulation should provide an incentive to generate better data and recommend adopting a pragmatic, but scientifically better founded approach to mixture risk assessment. © 2013 Martin et al.; licensee BioMed Central Ltd.Oak Foundatio

Movable genetic elements and antibiotic resistance in enterococci

The enterococci possess genetic elements able to move from one strain to another via conjugation. Certain enterococcal plasmids exhibit a broad host range among gram-positive bacteria, but only when matings are performed on solid surfaces. Other plasmids are more specific to enterococci, transfer efficiently in broth, and encode a response to recipient-produced sex phermones. Transmissible non-plasmid elements, the conjugative transposons, are widespread among the enterococci and determine their own fertility properties. Drug resistance, hemolysin, and bacteriocin determinants are commonly found on the various transmissible enterococcal elements. Examples of the different systems are discussed in this review.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47900/1/10096_2005_Article_BF01963632.pd

Simulating Microdosimetry in a Virtual Hepatic Lobule

The liver plays a key role in removing harmful chemicals from the body and is therefore often the first tissue to suffer potentially adverse consequences. To protect public health it is necessary to quantitatively estimate the risk of long-term low dose exposure to environmental pollutants. Animal testing is the primary tool for extrapolating human risk but it is fraught with uncertainty, necessitating novel alternative approaches. Our goal is to integrate in vitro liver experiments with agent-based cellular models to simulate a spatially extended hepatic lobule. Here we describe a graphical model of the sinusoidal network that efficiently simulates portal to centrilobular mass transfer in the hepatic lobule. We analyzed the effects of vascular topology and metabolism on the cell-level distribution following oral exposure to chemicals. The spatial distribution of metabolically inactive chemicals was similar across different vascular networks and a baseline well-mixed compartment. When chemicals were rapidly metabolized, concentration heterogeneity of the parent compound increased across the vascular network. As a result, our spatially extended lobule generated greater variability in dose-dependent cellular responses, in this case apoptosis, than were observed in the classical well-mixed liver or in a parallel tubes model. The mass-balanced graphical approach to modeling the hepatic lobule is computationally efficient for simulating long-term exposure, modular for incorporating complex cellular interactions, and flexible for dealing with evolving tissues

River Restoration in Spain: Theoretical and Practical Approach in the Context of the European Water Framework Directive.

River restoration is becoming a priority in many countries because of increasing the awareness of environmental degradation. In Europe, the EU Water Framework Directive (WFD) has significantly reinforced river restoration, encouraging the improvement of ecological status for water bodies. To fulfill the WFD requirements, the Spanish Ministry of the Environment developed in 2006 a National Strategy for River Restoration whose design and implementation are described in this paper. At the same time many restoration projects have been conducted, and sixty of them have been evaluated in terms of stated objectives and pressures and implemented restoration measures. Riparian vegetation enhancement, weir removal and fish passes were the most frequently implemented restoration measures, although the greatest pressures came from hydrologic alteration caused by flow regulation for irrigation purposes. Water deficits in quantity and quality associated with uncontrolled water demands seriously affect Mediterranean rivers and represent the main constraint to achieving good ecological status of Spanish rivers, most of them intensively regulated. Proper environmental allocation of in-stream flows would need deep restrictions in agricultural water use which seem to be of very difficult social acceptance. This situation highlights the need to integrate land-use and rural development policies with water resources and river management, and identifies additional difficulties in achieving the WFD objectives and good ecological status of rivers in Mediterranean countries

Mapping changing distributions of dominant species in oil-contaminated salt marshes of Louisiana using imaging spectroscopy

The April 2010 Deepwater Horizon (DWH) oil spill was the largest coastal spill in U.S. history. Monitoring subsequent change in marsh plant community distributions is critical to assess ecosystem impacts and to establish future coastal management priorities. Strategically deployed airborne imaging spectrometers, like the Airborne Visible/Infrared Imaging Spectrometer (AVIRIS), offer the spectral and spatial resolution needed to differentiate plant species. However, obtaining satisfactory and consistent classification accuracies over time is a major challenge, particularly in dynamic intertidal landscapes.Here, we develop and evaluate an image classification system for a time series of AVIRIS data for mapping dominant species in a heavily oiled salt marsh ecosystem. Using field-referenced image endmembers and canonical discriminant analysis (CDA), we classified 21 AVIRIS images acquired during the fall of 2010, 2011 and 2012. Classification results were evaluated using ground surveys that were conducted contemporaneously to AVIRIS collection dates. We analyzed changes in dominant species cover from 2010 to 2012 for oiled and non-oiled shorelines.CDA discriminated dominant species with a high level of accuracy (overall accuracy=82%, kappa=0.78) and consistency over three imaging dates (overall2010=82%, overall2011=82%, overall2012=88%). Marshes dominated by Spartina alterniflora were the most spatially abundant in shoreline zones (â¤28m from shore) for all three dates (2010=79%, 2011=61%, 2012=63%), followed by Juncus roemerianus (2010=11%, 2011=19%, 2012=17%) and Distichlis spicata (2010=4%, 2011=10%, 2012=7%).Marshes that were heavily contaminated with oil exhibited variable responses from 2010 to 2012. Marsh vegetation classes converted to a subtidal, open water class along oiled and non-oiled shorelines that were similarly situated in the landscape. However, marsh loss along oil-contaminated shorelines doubled that of non-oiled shorelines. Only S. alterniflora dominated marshes were extensively degraded, losing 15% (354,604m2) cover in oiled shoreline zones, suggesting that S. alterniflora marshes may be more vulnerable to shoreline erosion following hydrocarbon stress, due to their landscape position

The evolution of plasmid-carried antibiotic resistance

BACKGROUND: Antibiotic resistance represents a significant public health problem. When resistance genes are mobile, being carried on plasmids or phages, their spread can be greatly accelerated. Plasmids in particular have been implicated in the spread of antibiotic resistance genes. However, the selective pressures which favour plasmid-carried resistance genes have not been fully established. Here we address this issue with mathematical models of plasmid dynamics in response to different antibiotic treatment regimes. RESULTS: We show that transmission of plasmids is a key factor influencing plasmid-borne antibiotic resistance, but the dosage and interval between treatments is also important. Our results also hold when plasmids carrying the resistance gene are in competition with other plasmids that do not carry the resistance gene. By altering the interval between antibiotic treatments, and the dosage of antibiotic, we show that different treatment regimes can select for either plasmid-carried, or chromosome-carried, resistance. CONCLUSIONS: Our research addresses the effect of environmental variation on the evolution of plasmid-carried antibiotic resistance

Antibiotic resistance determinants in the interplay between food and gut microbiota

A complex and heterogeneous microflora performs sugar and lactic acid fermentations in food products. Depending on the fermentable food matrix (dairy, meat, vegetable etc.) as well as on the species composition of the microbiota, specific combinations of molecules are produced that confer unique flavor, texture, and taste to each product. Bacterial populations within such “fermented food microbiota” are often of environmental origin, they persist alive in foods ready for consumption, eventually reaching the gastro-intestinal tract where they can interact with the resident gut microbiota of the host. Although this interaction is mostly of transient nature, it can greatly contribute to human health, as several species within the food microbiota also display probiotic properties. Such an interplay between food and gut microbiota underlines the importance of the microbiological quality of fermented foods, as the crowded environment of the gut is also an ideal site for genetic exchanges among bacteria. Selection and spreading of antibiotic resistance genes in foodborne bacteria has gained increasing interest in the past decade, especially in light of the potential transferability of antibiotic resistance determinants to opportunistic pathogens, natural inhabitants of the human gut but capable of acquiring virulence in immunocompromised individuals. This review aims at describing major findings and future prospects in the field, especially after the use of antibiotics as growth promoters was totally banned in Europe, with special emphasis on the application of genomic technologies to improve quality and safety of fermented foods