Molecular approaches to the assessment of biodiversity in aquatic microbial communities

For the past 20 years, the increased development and routine application of molecular-based techniques has made it possible to carry out detailed evaluations of the biodiversity of aquatic microbial communities. It also offers great opportunities for finding out how this parameter responds to various environmental stresses. Most of these approaches involve an initial PCR amplification of a target, which is generally located within the ribosomal operon. The amplification is achieved by means of primers that are specific to the organisms of interest. The second step involves detecting sequence variations in the PCR fragments either by a cloning/sequencing analysis, which provides a complete characterization of the fragments, or by an electrophoretic analysis, which provides a visual separation of the mixture of fragments according to sequence polymorphism (denaturing or temperature gradient gel electrophoresis, single strand conformation polymorphism) or length polymorphism (terminal-restriction fragment length polymorphism, automated ribosomal intergenic spacer analysis). Other non-PCR-based methods are also commonly used, such as fluorescence in-situ hybridization and DNA re-association analysis. Depending on the technique used, the information gained can be quite different. Moreover, some of these analyses may be rather onerous in terms of time and money, and so not always suitable for screening large numbers of samples. The most widely used techniques are discussed in this paper to illustrate the principles, advantages and shortcomings of each of them. Finally, we will conclude by evaluating the techniques and discussing some emerging molecular techniques, such as real-time PCR and the microarray technique.     

Pine forest and grassland differently influence the response of soil microbial communities to metal contamination

Metal pollution can affect soil microbial communities, and vegetation potentially influences this relationship. It can, for example, modify the toxicity of metal to soil microbes by controlling its input to the ground or by altering soil physicochemical properties. This study examined metal effects on soil respiration, potentially active microbial biomass (SIR) and catabolic abilities of culturable heterotrophic bacterial communities (Biolog GN) in pine forest and grassland ecosystems developed on soils contaminated with Zn, Pb and Cd. In samples from non-forested areas we found that metal pollution reduced the microbial biomass and functional diversity of bacteria, while increasing the metabolic quotient. In samples from pine forests we found no relationship between metal pollution and microbial parameters. Metals induced changes in soil respiration neither in forest nor in grassland sites. Generally, microbial performance was determined predominantly by soil physicochemical properties (nutrient content, acidity, contamination level). Vegetation type seemed a minor but important factor influencing microbial communities. More work is needed to determine why even relatively high metal concentrations do not significantly affect microbial communities in forest soils.     

Assessment of changes in the microbial community of constructed wetland mesocosms in response to acid mine drainage exposure

Changes in the bacterial community in the interstitial water of 5 different constructed wetland mesocosms were studied over a 22-day period following exposure to simulated acid mine drainage (AMD). The community-level physiological profile (CLPP) of each mesocosm was assessed using substrate utilization patterns gathered via BIOLOG ECO plates. Principal component analysis (PCA) of the BIOLOG ECO plate data proved feasible and useful in characterizing the interstitial bacterial community in the constructed wetland mesocosms based on mesocosm characteristics such as fixed biological regime development and plant presence, and was also used to successfully track changes in the interstitial bacterial community in response to AMD exposure. Clustering analysis of the BIOLOG ECO plate data was used to characterize the interstitial bacterial community and to validate the mesocosm groupings observed through PCA ordination. The calculation of substrate-based diversity indices from the BIOLOG ECO plate data was used to assess the robustness and the degree of change shown by the metabolic fingerprint of the interstitial bacterial community within the mesocosms. The interstitial bacterial community in the constructed mesocosms was shown to be significantly affected after exposure to AMD. Exposure to AMD caused similar bacterial species to detach from the fixed biotic regime of the mesocosms. It was also shown that mesocosms planted with Phragmites australis did not experience as great an ecological shift in the microbial ecology of the interstitial water after exposure to AMD as did the unplanted mesocosms. The substrate-based diversity indices for the planted mesocosms were also found to be more stable, after exposure to AMD, than those for the unplanted mesocosms. It is possible that the interaction between the plant root system and the substrate biological regime, collectively called the rhizosphere, may create a more ecologically robust and stable treatment system.     

Toxicity and potential risk assessment of a river polluted by acid mine drainage in the Iberian Pyrite Belt (SW Spain)

Metal contamination from acid mine drainage (AMD) is a serious problem in the southwest of the Iberian Peninsula, where the Iberian Pyrite Belt is located. This zone contains original sulfide reserves of about 1700 Mt distributed among more than 50 massive sulfide deposits. Weathering of these minerals releases to the waters significant quantities of toxic elements, which severely affect the sediments and surface waters of the region. The main goal of this paper is to evaluate the toxicity and the potential risk associated with the mining areas using Microtox test and different factors which assess the degree of contamination of the sediments and waters. For this, a natural stream polluted by AMD-discharge from an abandoned mine has been studied. The results show that elevated concentrations of Cu, As and Zn involve an important potential risk on the aquatic environment, associated both with sediments and waters. Microtox test informs that the sediments are extremely or very toxic, mainly related to concentrations of Fe, As, Cr, Al, Cd, Cu and Zn. Pollution is mainly transferred to the sediments increasing their potential toxicity. A natural creek affected by AMD can store a huge amount of pollution in its sediments while exhibiting a not very low water pH and low water metal concentration.     

An assessment of the sensitivity of lattice towers to fatigue induced by wind gusts

The dynamic properties of lattice towers are closely constrained by practical economic design. It is thus possible to generalize the prediction of dynamic response, using the stochastic wind gust model, with details as outlined in the seminar ‘Wind engineering in the eighties’ (CIRIA, London, 1980). An approximation to the combined effect of resonant and non-resonant components of the response is presented, as a factor to be applied to the effective stress range based on the dynamic response alone, computed at a single ‘reference wind speed’.

Results are presented for a range of practical structures, which indicate that good detailing can generally ensure that fatigue damage caused by wind gusts does not seriously limit design. The sensitivity of the normalized results to changes of location and of tower function and geometry is shown to be relatively small.