Remote mountain lakes as indicators of diffuse acidic and organic pollution in the Iberian peninsula (AL:PE 2 studies)

In the framework of the AL:PE 2 project, studies on acidification and organic pollution in mountain lakes have been conducted in several ranges in the Iberian peninsula: Pyrenees (Northeastern Spain), Sierra de Gredos (Central Spain), Sierra Nevada (Southern Spain) and Serra da Estrela (Central Portugal). These studies focused on water and sediment chemistry and organisms (benthic diatoms, Zooplankton, aquatic macroinvertebrates, and fish) as indicators of acidification. Organic micropollutants (PAH, PCB, DDE, hexachlorobenzene and others) in lake sediments and fish have been studied as tracers of atmospheric pollution. The Iberian peninsula lakes do not show severe anthropogenic acidification. pH values are in the range of sensitive lakes, but the levels of acidic pollutants are low. The status of the organisms surveyed agreed with this diagnosis. Pyrenean lakes showed the highest fluxes of organic pollutants related to fossil fuel combustion., higher pollutioninduced versus natural acidity ratios, and modeled alkalinity and pH declines.     

Application of relative quantification TaqMan real-time polymerase chain reaction technology for the identification and quantification of Thunnus alalunga and Thunnus albacares

A novel one-step methodology based on real-time Polymerase Chain Reaction (PCR) technology has been developed for the identification of two of the most valuable tuna species. Nowadays, species identification of seafood products has a major concern due to the importing to Europe of new species from other countries. To achieve this aim, two specific TaqMan systems were devised to identify Thunnus alalunga and Thunnus albacares. Another system specific to Scombroidei species was devised as a consensus system. In addition, a relative quantification methodology was carried out to quantify T. alalunga and T. albacares in mixtures after the relative amount of the target was compared with the consensus. This relative quantification methodology does not require a known amount of standard, allowing the analysis of many more samples together and saving costs and time. The utilization of real-time PCR does not require sample handling, preventing contamination and resulting in much faster and higher throughput results.     

Isolation and thermal characterization of an acidic isoperoxidase from turnip roots

An acidic peroxidase (pI approximately 2.5) was purified from turnip roots (TAP), and its thermal properties were evaluated. TAP is a monomeric protein having a molecular weight (MW) of 49 kDa and a carbohydrate content accounting for 18% of the MW. The yield of pure TAP was relatively high ( approximately 2 mg/kg of fresh roots), with a specific activity of 1810 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) units/mg at pH 6. The activity increased 4-fold at the optimum pH (4.0) to 7250 ABTS units/mg, higher than that of most peroxidases. TAP was heat stable; heat treatment of 25 min at 60 degrees C resulted in 90% initial activity retention, whereas an activity of 20% was retained after 25 min of heating at 80 degrees C. TAP regained 85% of its original activity within 90 min of incubation at 25 degrees C, following heat treatment at 70 degrees C for 25 min. Thermal inactivation caused noticeable changes in the heme environment as evaluated by circular dichroism and visible spectrophotometry. TAP was rapidly denatured by heating in the presence of 1.0 mM ethylene glycol bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid, but the Soret band and activity were fully recovered by adding an excess of Ca(2+). This is further evidence that Ca(2+) plays an important role in the stability of TAP. The high specific activity of TAP, together with its relatively high thermal stability, has high potential for applications in which a thermally stable enzyme is required.     

Soil physical properties and soybean (Glycine max, Merrill) root abundance in conventionally- and zero-tilled soils in the humid Pampas of Argentina

In the humid Pampas of Argentina soybean is cultivated in different soil types, which were changed from conventional- to zero tillage systems in the last decade. Little is known about the response of soybean roots to these different soil physical environments. Pasture, and conventionally- and zero-tilled field lots cropped to soybean (R1 and R2 ontogenic stages) were sampled in February–March 2001 in a sandy clay loam and two silty clay loam Mollisols, and in a clayey Vertisol. In the 0–0.05 m layer of conventionally- and zero-tilled lots soil organic carbon represented 53–72% of that in pasture lots, and showed an incipient recovery after 4–11 years of continuous zero tillage. Soil aggregate stability was 10.1–46.8% lower in conventionally-tilled than in pasture lots, and recovered completely in zero-tilled lots. Soil relative compaction ranged 60.8–83.6%, which was below the threshold limit for crop yields (>90%). In change, soil porosity >50 μm ranged 0.91–5.09% soil volume, well below the minimum critical limit for root aeration and elongation (>10%, v/v). The threshold of soil resistance (about 2–3 MPa) was only over passed in an induced plough pan in the conventionally-tilled Bragado soil (5.9 MPa), and in the conventionally- and zero-tilled Ramallo soils (3.7–4.2 MPa, respectively). However, neither the low macroporosity nor the high soil resistances impeded soybean roots growth in any site. According to a fitted polynomial function, root abundance was negatively related to clay content in the subsoil (R² = 0.84, P < 0.001). Soybean roots were only abundant in the subsoil of the sandy clay loam Mollisol, which had <350 g kg⁻¹ clay. Results show that subsoil properties, and not tillage systems, were the primary effect of root growth of soybean.     

Rabbits as a keystone species in southern Europe

A keystone species is one that is crucial in maintaining the organization and diversity of their ecological communities. We tested the idea that rabbits act as a keystone species in southern Europe by exploring relationships between rabbit abundance and the abundance and diversity of raptor species. At 20 sites in southern Spain we assessed rabbit abundance through counts of animals along transects and assessed the number of raptor individuals and species through watches from vantage points. In a further 120 locations we also derived an index of rabbit abundance, from pellets and compared this to the presence or absence of the critically endangered Spanish imperial eagle. Rabbit abundance was positively associated with the number of observations of raptors, the number of raptor species and the number of species of conservation concern. Sites with the most rabbits had higher conservation value. Moreover, the presence of Spanish imperial eagle was strongly associated with sites where rabbits were at high density. We conclude that rabbits do act as a keystone species and we suggest that conservation efforts should focus on improving the status of this small-game species in southern Europe.     

Identification of goose,mule duck,chicken, turkey,and swine in foie gras by species-specific polymerase chain reaction

A specific Polymerase Chain Reaction (PCR) has been developed for the identification of goose (Anser anser), mule duck (Anas platyrhynchos x Cairina moschata), chicken (Gallus gallus), turkey (Meleagris gallopavo), and swine (Sus scrofa domesticus) in foie gras. A forward common primer was designed on a conserved DNA sequence in the mitochondrial 12S ribosomal RNA gene (rRNA), and reverse primers were designed to hybridize on species-specific DNA sequences of each species considered. The different sizes of the species-specific amplicons, separated by agarose gel electrophoresis, allowed clear identification of goose, mule duck, chicken, turkey, and swine in foie gras. Analysis of experimental mixtures demonstrated that the detection limit of the assay was approximately 1% for each species analyzed. This genetic marker can be very useful for the accurate identification of these species, avoiding mislabeling or fraudulent species substitution in foie gras.     

Incorporation of trace elements on iron-rich concretions around plant roots of tagus estuary salt marsh (portugal)

Objective

The concentration gradients of solid Al, Fe, Mn, Zn, Cd, Pb, Cr and Ni in freshly formed concretions and in older concretions preserved in the sediment were evaluated in a millimetre resolution scale. These results provide a better understanding of the complex biogeochemical processes in the root-sediment system and elucidate the potentialities of marshes in the restoration of contaminated aquatic systems.

Methods

Tens (soll das vielleicht tonnes/tons heißen) of freshly formed concretions and preserved concretions were collected in the Rosário salt marsh. Each tubular structure and involving sediments were scraped in concentric layers of 2-mm thickness to form two sets of composite samples. The elemental composition of concretion and sediment samples were determined by atomic absorption spectrometry.

Results and Discussion

The sediment involving the concretions contained ～0.5 mmol g^-1 Fe, but concentrations increased towards the root up to 3.4 mmol g^-1 in 3-mm interval. Manganese, Zn, Cd and Pb exhibited similar radial distribution in both preserved and freshly formed concretions indicating that these metals remain in those structures with the age. Chromium and Ni showed a different pattern being lower in concretions with respect to involving sediments, and consequently, concretions do not act as a barrier for these elements. The estimated amount of Zn (59 tons), Pb (5 tons) and Cd (0.4 tons) retained in the concretions all over the Rosario salt marsh highlights the importance of these structures in sequestering metals.

Conclusions and Outlook

The sequestering may be particularly relevant in salt marshes located near urban and industrial zones, being areas working as a natural remediation system and contributing for the detoxification of aquatic environments and the well-being of coastal resources.

     

Treatment of metastatic melanoma B16F10 by the flavonoids tangeretin, rutin, and diosmin

Melanoma is one of the most frequently metastasizing malignant neoplasias. This study examines an experimental model of pulmonary metastasis and the B16F10 cell subline, highly metastatic in the lung. Antimetastatic effects of the flavonoids tangeretin, rutin, and diosmin were analyzed, and at the same time an analysis of the metastatic activity of ethanol was performed, considered to be necessary because it is used as a vehicle for administering the flavonoids. Lentini's model, which complements the macroscopic evaluation of nodule numbers by using a stereoscopic microscope and image analysis at the microscopic level, was used. The greatest reduction in the number of metastatic nodules (52%) was obtained with diosmin; similarly, the percentages of implantation, growth index, and invasion index (79.40, 67.44, and 45.23%, respectively), were all compared with those of the ethanol group, considered to be an effective control group. Rutin- and tangeretin-treated groups also showed reductions of the same index compared with the ethanol group. It would seem that structural factors would better explain these results and the antimetastatic activity of each flavonoid and the respective metabolites.