Evaluation of concentration efficiency of the Pseudomonas aeruginosa phage PP7 in various water matrixes by different methods

Enteric viruses monitoring in surface waters requires the concentration of viruses before detection assays. The aim of this study was to evaluate different methods in terms of recovery efficiencies of bacteriophage PP7 of Pseudomonas aeruginosa, measured by real-time PCR, using it as a viral control process in water analysis. Different nucleic acid extraction methods (silica–guanidinium thiocyanate, a commercial kit (Qiagen Viral RNA Kit) and phenol–chloroform with alcohol precipitation) exhibited very low recovery efficiencies (0.08–4.18 %), being the most efficient the commercial kit used for subsequent experiments. To evaluate the efficiency of three concentration methods, PBS (as model for clean water) and water samples from rivers were seeded to reach high (HC, 10⁶ pfu ml^?1) and low concentrations (LC, 10⁴ pfu ml^?1) of PP7. Tangential ultrafiltration proved to be more efficient (50.36?±?12.91, 17.21?±?9.22 and 12.58?±?2.35 % for HC in PBS and two river samples, respectively) than adsorption–elution with negatively charged membranes (1.00?±?1.34, 2.79?±?2.62 and 0.05?±?0.08 % for HC in PBS and two river samples, respectively) and polyethylene glycol precipitation (15.95?±?7.43, 4.01?±?1.12 and 3.91?±?0.54 %, for HC in PBS and two river samples, respectively), being 3.2–50.4 times more efficient than the others for PBS and 2.7–252 times for river samples. Efficiencies also depended on the initial virus concentration and aqueous matrixes composition. In consequence, the incorporation of an internal standard like PP7 along the process is useful as a control of the water concentration procedure, the nucleic acid extraction, the presence of inhibitors and the variability of the recovery among replicas, and for the calculation of the sample limit of detection. Thus, the use of a process control, as presented here, is crucial for the accurate quantification of viral contamination.     

Competitive kinetics of adsorption onto activated carbon for emerging contaminants with contrasting physicochemical properties

Environmental Science and Pollution Research - Activated carbon (AC) can be used for the removal of emerging contaminants (e.g., drugs) in water and wastewater treatment plants. In the present...     

Estimates of residual dairy manure nitrogen availability using various techniques

It is common practice to repeatedly apply dairy manure to the same fields. To accurately assess the total plant availability of manure nutrients, it is necessary to account for the nutrients remaining in soil from previous manure applications. A field experiment studying manure nitrogen (N) uptake by corn (Zea mays L.) was conducted from 1998 to 2003 on a Plano silt loam (fine-silty, mixed, mesic, Typic Argiudolls). Plots received two rates of semisolid manure either every year, every 2 yr, or every 3 yr to estimate first-, second-, and third-year dairy manure N residuals. Residual manure N availability was estimated from single and multiple manure applications using (i) the fertilizer N equivalence method, (ii) the apparent recovery (difference) method, (iii) relative effectiveness method, and (iv) recovery of (15)N-labeled manure. Second-year availabilities after a single manure application using the fertilizer equivalence, difference, and relative effectiveness methods were estimated to be 12, 8, and 4% of total manure N applications, respectively. Estimates of third-year availability by these methods were 3, 1, and 5%, respectively. Measurement of (15)N recovered from labeled manure was 6 and 2% in the second and third year, respectively. Fertilizer equivalence, difference, and relative effectiveness methods showed great year to year variability, reducing the confidence in the residual manure N availability estimates by these methods, but using (15)N-labeled manures reduced variability substantially. Based on this and other studies, we suggest that second- and third-year residual N availability from a single application of semisolid dairy manure would be 9 to 12%, and 3 to 5% of the original manure N application, respectively.     

Sustainability: A Study of the Level of Ecological Behavior of Postgraduate Students in Brazil

The decline in the supply of natural resources is a cause for concern, especially given continued worldwide population growth. New habits and attitudes toward resource use are slowly being introduced into the everyday lives of persons residing in a number of countries. The term “ecological behavior” refers to acting in favor of the environment, whether these actions are voluntarily or not, or learned or not (Pato & Tamayo, 2006 ). The general objective of this study is to identify the environmental behavior—one of several terms used interchangeably with ecological behavior—of graduate students from a Brazilian federal institution of higher education. We chose this particular group because many of them already act, or will act, as professors. Thus, by virtue of their positions, they will function as opinion leaders who will influence the education of future professionals, establishing a chain through which shared values can gradually change the environmental behavior of society.     

Using lysosomal membrane stability of haemocytes in Ruditapes philippinarum as a biomarker of cellular stress to assess contamination by caffeine, ibuprofen, carbamazepine and novobiocin

Although pharmaceuticals have been detected in the environment only in the range from ng/L to g/L, it has been demonstrated that they can adversely affect the health status of aquatic organisms. Lysosomal membrane stability (LMS) has previously been applied as an indicator of cellular well-being to determine health status in bivalve mussels. The objective of this study is to evaluate LMS in Ruditapes philippinarum haemolymph using the neutral red retention assay (NRRA). Clams were exposed in laboratory conditions to caffeine (0.1, 5, 15, 50 μg/L), ibuprofen (0.1, 5, 10, 50 μg/L), carbamazepine and novobiocin (both at 0.1, 1, 10, 50 μg/L) for 35 days. Results show a dose-dependent effect of the pharmaceuticals. The neutral red retention time measured at the end of the bioassay was significantly reduced by 50% after exposure to environmental concentrations (p < 0.05) (caffeine = 15 μg/L; ibuprofen = 10 μg/L; carbamazepine = 1 μg/L and novobiocin = 1 μg/L), compared to controls. Clams exposed to these pharmaceuticals were considered to present a diminished health status (retention time < 45 min), significantly worse than controls (96 min) (p < 0.05). The predicted no environmental effect concentration (PNEC) results showed that these pharmaceuticals are very toxic at the environmental concentrations tested. Measurement of the alteration of LMS has been found to be a sensitive technique that enables evaluation of the health status of clams after exposure to pharmaceuticals under laboratory conditions, thus representing a robust Tier-1 screening biomarker.     

Levels of plasma B-esterases and glutathione-S-transferase activities in three South American toad species

We determined the normal levels of butyrylcholinesterase (BChE), carboxylesterase (CbE), and glutathione S-transferases (GST) activities in three South American toad species in order to establish reference values for field pesticide monitoring purposes. Interspecies variations in B-esterase and GST activities were examined according to body mass. In addition, comparative inhibition of BChE and CbE activities using malaoxon, and chemical reactivation of malaoxon-inhibited BChE activity using pyridine-2-aldoxime methochloride (2-PAM) were investigated. Bufo fernandezae had average activity values for BChE: 17.31 mmol min^?1 ml^?1; CbE: 621.49 nmol min^?1 ml^?1 and GST: 1.94 mmol min^?1 ml^?1 while B. arenarum enzymatic average activities were BChE: 9.51 mmol min^?1 ml^?1; CbE: 270.07 nmol min^?1 ml^?1, and GST: 1.59 mmol min^?1 ml^?1; finally Bufo schneideri had enzymatic mean values of BChE: 2.08 mmol min^?1 ml^?1; CbE: 301.95 nmol min^?1 ml^?1, and GST: 1.60 mmol min^?1 ml^?1. Moreover, we found an allometric relationship between plasma BChE and CbE activities and body size for the three toad species. We suggest that B. fernandezae would be the species with a higher tolerance capacity to organophosphorous insecticides compared to the other toad species, while B. schneideri may be the most vulnerable toad species to field pesticide exposure, although some other factors (e.g., brain AChE sensitivity or pesticide metabolism by phosphotriesterases) should be also taken into account. The malaoxon-inhibited BChE activity of the three toad species was reactivated in the presence of 2-PAM, and it is recommended as a specific and sensitive methodology in the assessment of field exposure to OP insecticides together to compare BChE activity levels between OP-exposed and nonexposed individuals.     

The reproduction of Enchytraeus sp.—technical improvement for the counting of juveniles

Soil-dwelling annelids of the genus Enchytraeus are used in ecotoxicology for testing of chemicals mixed in artificial soil or for testing of wastes and soils of unknown quality. ISO 16387 describes a method for determining the effects of substances or contaminated soils on survival and reproduction of Enchytraeus albidus or of the smaller species Enchytraeus buchholzi or Enchy-traeus crypticus. After the total test duration of 6 (or 4)?weeks, the juveniles hatched in the meantime are counted. There are several possible extraction techniques, which are always followed by counting the juveniles by hand, but none of them seems easy to handle. We proposed a new modification of the worm extraction method using flotation of fixed and stained juveniles followed by taking a photograph. The digital image of the juveniles is evaluated by computer processing. It makes the counting of juveniles much easier and less labor intensive.     

Baselines for land-use change in the tropics: application to avoided deforestation projects

Although forest conservation activities, particularly in the tropics, offer significant potential for mitigating carbon (C) emissions, these types of activities have faced obstacles in the policy arena caused by the difficulty in determining key elements of the project cycle, particularly the baseline. A baseline for forest conservation has two main components: the projected land-use change and the corresponding carbon stocks in applicable pools in vegetation and soil, with land-use change being the most difficult to address analytically. In this paper we focus on developing and comparing three models, ranging from relatively simple extrapolations of past trends in land use based on simple drivers such as population growth to more complex extrapolations of past trends using spatially explicit models of land-use change driven by biophysical and socioeconomic factors. The three models used for making baseline projections of tropical deforestation at the regional scale are: the Forest Area Change (FAC) model, the Land Use and Carbon Sequestration (LUCS) model, and the Geographical Modeling (GEOMOD) model. The models were used to project deforestation in six tropical regions that featured different ecological and socioeconomic conditions, population dynamics, and uses of the land: (1) northern Belize; (2) Santa Cruz State, Bolivia; (3) Paraná State, Brazil; (4) Campeche, Mexico; (5) Chiapas, Mexico; and (6) Michoacán, Mexico. A comparison of all model outputs across all six regions shows that each model produced quite different deforestation baselines. In general, the simplest FAC model, applied at the national administrative-unit scale, projected the highest amount of forest loss (four out of six regions) and the LUCS model the least amount of loss (four out of five regions). Based on simulations of GEOMOD, we found that readily observable physical and biological factors as well as distance to areas of past disturbance were each about twice as important as either sociological/demographic or economic/infrastructure factors (less observable) in explaining empirical land-use patterns. We propose from the lessons learned, a methodology comprised of three main steps and six tasks can be used to begin developing credible baselines. We also propose that the baselines be projected over a 10-year period because, although projections beyond 10 years are feasible, they are likely to be unrealistic for policy purposes. In the first step, an historic land-use change and deforestation estimate is made by determining the analytic domain (size of the region relative to the size of proposed project), obtaining historic data, analyzing candidate baseline drivers, and identifying three to four major drivers. In the second step, a baseline of where deforestation is likely to occur–a potential land-use change (PLUC) map—is produced using a spatial model such as GEOMOD that uses the key drivers from step one. Then rates of deforestation are projected over a 10-year baseline period based on one of the three models. Using the PLUC maps, projected rates of deforestation, and carbon stock estimates, baseline projections are developed that can be used for project GHG accounting and crediting purposes: The final step proposes that, at agreed interval (e.g., about 10 years), the baseline assumptions about baseline drivers be re-assessed. This step reviews the viability of the 10-year baseline in light of changes in one or more key baseline drivers (e.g., new roads, new communities, new protected area, etc.). The potential land-use change map and estimates of rates of deforestation could be re-done at the agreed interval, allowing the deforestation rates and changes in spatial drivers to be incorporated into a defense of the existing baseline, or the derivation of a new baseline projection.     

Response of leaf and fine roots proteomes of Salix viminalis L. to growth on Cr-rich tannery waste

Environmental Science and Pollution Research - The tannery industry is a major source of anthropogenic chromium (Cr) contamination due to the large amounts of solid waste produced and its...