Diversity and community structure of culturable arsenic-resistant bacteria across a soil arsenic gradient at an abandoned tungsten-tin mining area

We studied the bacterial diversity at a single location (the Terrubias mine; Salamanca province, Spain) with a gradient of soil As contamination to test if increasing levels of As would (1) change the preponderant groups of arsenic-resistant bacteria and (2) increase the tolerance thresholds to arsenite [As(III)] and arsenate [As(V)] of such bacteria. We studied the genetic and taxonomic diversity of culturable arsenic-resistant bacteria by PCR fingerprinting techniques and 16S rRNA gene sequencing. Then, the tolerance thresholds to As(III) and As(V) were determined for representative strains and mathematically analyzed to determine relationships between tolerances to As(III) and As(V), as well as these tolerances with the soil contamination level. The diversity of the bacterial community was, as expected, inversely related to the soil As content. The overall preponderant arsenic-resistant bacteria were Firmicutes (mainly Bacillus spp.) followed by γ-Proteobacteria (mainly Pseudomonas spp.), with increasing relative frequencies of the former as the soil arsenic concentration increased. Moreover, a strain of the species Rahnella aquatilis (γ-Proteobacteria class) exhibited strong endurance to arsenic, being described for the first time in literature such a phenotype within this bacterial species. Tolerances of the isolates to As(III) and As(V) were correlated but not with their origin (soil contamination level). Most of the strains (64%) showed relatively low tolerances to As(III) and As(V), but the second most numerous group of isolates (19%) showed increased tolerance to As(III) rather than to As(V), even though the As(V) anion is the prevalent arsenic species in soil solution at this location. To our knowledge, this is the first study to report a shift towards preponderance of Gram-positive bacteria (Firmicutes) related to high concentrations of soil arsenic. It was also shown that, under aerobic conditions, strains with relatively enhanced tolerance to As(III) predominated over the most As(V)-tolerant ones.     

Effects of cold-dark storage on growth of Cylindrotheca closterium and its sensitivity to copper

Cylindrotheca closterium cells were maintained at low temperature (4+/-1 degrees C) and dark conditions up to 21 weeks to assess the effect on survival and physiological status. From a control culture under standard conditions, three densities were prepared: (A) 2 x 10(4), (B) 10 x 10(4), and (C) 25 x 10(4) cells ml(-1). Weekly, inoculums of each stored density were exposed to continuous light and at 20+/-1 degrees C. Sensitivity to copper for microalgal cultures was evaluated in order to assess possible changes in cells sensitivity due to storage. Concurrently, assays with a control culture were carried out in order to assess the sensitivity of C. closterium to copper and to be able to generate a standard sensitivity control chart with a mean value of EC50-72 h+/-2SD (standard deviation). Density-C presented higher cell yield values, between 40% and 80% relative to control culture. Cell density showed to be important feature that may be taken into account in cell storage experiments. There was an increase in sensitivity of cells submitted to storage; however results always kept in the range established as standard sensitivity with no statistically significant difference with regards to control culture. EC50-72 h mean value for the control culture was 29+/-10 mug Cul(-1), while for densities-A, B and C were 22+/-7; 23+/-9 and 23+/-8 microg Cul(-1), respectively. In spite of drastic changes in the environmental conditions due to storage, it is concluded that C. closterium cells stored during 5 months remained metabolically active and with no significant change in its sensitivity.     

Spatial and temporal trends of PCDDs and PCDFs in bivalve mollusc coming from Galicia (2000-2005). Possible relationship between biometric parameters and PCDDs and PCDFs levels

Levels and specific profiles of PCDD/F congeners were determined in bivalve mollusc coming from several Rías (estuarine bays) in Galicia (Spanish northwest Atlantic coast). Three species of bivalve mollusc, Mytilus galloprovincialis, Cerstoderma edulis and Ostrea edulis, from several points of littoral collected in the period from 2000 to 2005 were analysed. WHO-TEQ concentrations ranged from 0.08 to 1.62pgg(-1) wet weight, values below the maximum concentration established by the EU. The PCDD/PCDF congeners profile in the studied samples was dominated by 2,3,7,8-TCDF and 2,3,4,7,8-PeCDF. The possible influence between biometric parameters (fat and condition index) and the culture type of mussel (wild or raft) on PCDDs and PCDFs levels were studied using statistical analysis. The coefficients of Pearson product-moment correlation indicated the existence of positive significant relationship between 2,3,7,8-TCDF and 2,3,4,7,8-PeCDF levels and fat content. There was not correlation between condition index and PCDD/Fs content. ANOVA also revealed there were significant differences between summation operatorPCDD/Fs, 2,3,7,8-TCDF and 2,3,4,7,8-PeCDF levels in raft and wild mussels. Spatial and temporal trends were supported by patterns in ANOVA, using a general linear model (GLM) showing a clear separation in the composition of these compounds in more and less contaminated Rías and a decrease of PCDD/FS levels along the years.     

Performance of <Emphasis Type="Italic">Eleocharis macrostachya</Emphasis> and its importance for arsenic retention in constructed wetlands

Introduction  

Arsenic (As) can be removed from water via rhizofiltration using phytostabilizing plants. The aim of this study was to investigate the performance of Eleocharis macrostachya in constructed wetland prototypes, as well as the plant's arsenic mass retention and the distribution of As along the wetland flow gradient and the soil in the wetland mesocosmos.     

Temperature enhanced effects of chlorine exposure on the health status of the sentinel organism Mytilus galloprovincialis

It now is widely recognised that the global temperature is rising, a phenomenon which could alter the effects of pollution on wildlife. In order to assess the role of temperature and exposure to chlorine due to cooling water discharges, a battery of metabolic, oxidative stress and histological parameters were evaluated in Mytilus galloprovincialis after 15 and 30 days at 15 °C and at two increased temperatures (+5 and +10 °C). Diverse gill pathologies such as haemolymphatic sinus dilatation, an increased number of mucocytes and granulocytes as well as a lower number of cilia were observed after 30 days exposure at higher temperatures. Protein, amino acid, triglyceride and fatty acid levels decreased when the temperature increased, as a consequence of higher energetic demand. Similarly, acetylcholinesterase, catalase and glutathione S-transferase activities showed an inhibition at higher temperatures, although gill lipid peroxidation levels remained unaffected. Our results suggest that increased temperatures induce deterioration in the health status of the mussels and in their defensive capacity against a polluted environment.     

Association between Levels of Fine Particulate and Emergency Visits for Pneumonia and other Respiratory Illnesses among Children in Santiago,Chile

ABSTRACT

Recent evidence has implicated the fine fraction of particulate as the major contributor to the increase in mortality and morbidity related to particulate ambient levels. We therefore evaluated the impact of daily variation of ambient PM_2.5 and other pollutants on the number of daily respiratory-related emergency visits (REVs) to a large pediatric hospital of Santiago, Chile. The study was conducted from February 1995 to August 1996. Four monitoring stations from the network of Santiago provided air pollution data. The PM2.5 24-hr average ranged from 10 to 111 μg/m³ during September to April (warm months) and from 10 to 156 μg/m³ during May to August (cold months). Other contaminants (ozone (O₃), nitrogen dioxide (NO₂), and sulfur dioxide (SO₂)) were, in general, low during the study period. The increase in REVs was significantly related to PM₁₀ and PM_2.5 ambient levels, with the relationship between PM_2.5 levels and the number of REVs the stronger. During the cold months, an increase of 45 ìg/m³ in the PM_2.5 24-hr average was related to a 2.7% increase in the number of REVs (95% CI, 1.1–4.4%) with a two-day lag, and to an increase of 6.7% (95% CI, 1.7–12.0%) in the number of visits for pneumonia with a three-day lag. SO₂ and NO₂ were also related to REVs. We conclude that urban air pollutant mixture, particularly fine particulates, adversely affect the respiratory health of children residing in Santiago.     

Liquid chromatography-hydride generation-atomic absorption spectrometry for the speciation of tin in seafoods

Liquid chromatography with hydride generation atomic absorption spectrometry as the detection system was applied to the separation and determination of inorganic tin, tributyltin, dibutyltin, monobutyltin, diphenyltin and monophenyltin. A reversed phase C18 column and a methanol/water/acetic acid (70:27:3, v/v/v) mixture containing 0.05%(v/v) triethylamine and 0.1%(w/v) tropolone as the mobile phase (isocratic elution) were used. Extraction of organotins from the samples was carried out using methanol containing 0.05%(w/v) tropolone, a process that was repeated twice. The supernatants were shaken with water and dichloromethane in a separating funnel and the organic phase was collected and evaporated to dryness. When the method was applied to the speciation of tin in fresh and canned mussels, no organotins above the detection limits were identified in any of the samples, inorganic tin being the only species detected. The reliability of the procedure was checked by analyzing the total tin content of the samples by electrothermal atomic absorption spectrometry and by speciation of tin in a certified reference material, mussel tissue (CRM 477). The method can be used for environmental monitoring of organotins contaminated samples.     

Evaluation of porous cup soil‐water samplers under laboratory conditions: Comparison of ceramic and PTFE cups

Three different types of soil solution samplers (two ceramic cups and PTFE) were tested in the laboratory to validate their use for collecting soil‐water samples and for monitoring the chemical composition of soil solutions. Interactions between porous materials and chemical solutions were examined under different experimental conditions for several major cations (Ca²⁺, Mg²⁺, Na⁺ and K⁺) and anions (HCO^‐ ₃, NO^‐ ₃, SO₄ ^2‐ and Cl^‐) and for several minor ions (NH⁺ ₄, NO^‐ ₄, PO₄ ^3‐and B).

The results show that ceramic cups are not inert for bicarbonate, calcium and phosphate ions. Their use is only valid for a limited number of ions such as chloride and nitrate which, however, are the most studied ions in field experiments. PTFE cups have less restrictions although their use in measuring sulphates, phosphates and ammonium presents some problems.

Choosing the appropriate type of cup depends on the objectives. In any case, laboratory experiments must be performed before installation.     

Geographic patterns of natal dispersal in barn swallows <Emphasis Type="Italic">Hirundo rustica</Emphasis> from Denmark and Spain

Dispersal is an important factor that determines the degree of gene flow and, hence, the degree of differentiation among populations. Using two long-term datasets on natal philopatry and short-distance dispersal in barn swallows Hirundo rustica from Denmark and Spain, we evaluated the fitness costs and benefits and test a number of predictions about the functional significance of dispersal. The proportion of philopatric individuals was more than six times larger in Spain than in Denmark, with a higher rate of philopatry in males than in females. Dispersal propensity decreased in both populations during the course of the study. Males from the more philopatric Spanish population lived longer when philopatric rather than dispersing while that was not the case for either sex of the less philopatric Danish population. There were large differences in dispersal propensity among cohorts and breeding sites, suggesting that sites differed in their suitability as sites for immigrants. We found no evidence consistent with the mate competition hypothesis suggesting that males in better condition or with larger condition-dependent secondary sexual characters were more likely to be philopatric. These findings suggest that there is a high degree of intraspecific variation in dispersal propensity between populations, probably relating to local differences in costs and benefits of philopatry and dispersal.     

Sublethal toxicity of the Prestige oil spill on yellow-legged gulls

The Prestige oil spill in November 2002 is considered the biggest large-scale catastrophe of its type in Europe, thousands of seabirds dying in the subsequent months. Here, the total concentration of 16 polycyclic aromatic hydrocarbons (TPAH) was measured in the blood cell fraction of adult and chick yellow-legged gulls (Larus michahellis) from unoiled and oiled coastal areas in North Western Spain. In addition, hematocrit, plasma metabolites, electrolytes and enzymes, as well as body mass were determined in the same individuals. Our results strongly suggest the presence of health damages of sublethal nature in adult gulls breeding in oiled colonies 17 months after the Prestige oil spill. This is supported by the following evidences: (1) gulls sampled in unoiled and oiled colonies differed in blood TPAH levels, (2) gulls sampled in unoiled and oiled colonies differed in several blood parameters indicative of physiological disorders, and (3) TPAH in blood was significantly related to several of these parameters. Differences in the level of asparatate aminotransferase (AST), gamma-glutamyl transferase (GGT), total protein, glucose and inorganic phosphorus suggest damages on some vital organs (i.e. liver and kidney) in adult birds from oiled areas. Meanwhile, chicks presented weaker effects than adults, showing only between-area differences in hematocrit. Since TPAH levels in blood did not differ between both age-groups, the stronger effects on adults should be due to their longer exposure to these pollutants and/or to severe exposure in the months following the spill. The presence of PAHs in chicks indicates that these pollutants were incorporated into the food chain because nestlings would have been only exposed to contaminated organisms in the diet (e.g. fishes and crustaceans). Our findings support the view that PAHs may deeply alter the physiology of seabirds, and emphasize the necessity of quantifying the circulating levels of these compounds in order to evaluate the sublethal effects associated to large oil spills.