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The equity and efficiency principle in the financing system of the NHS in Portugal.

The public financing systems of primary health care services and hospital services in Portugal have traditionally been separated. In the United Kingdom where a similar separation exists, inefficient transfers of services take place from the hospital services to the primary care services. In this article a new way of financing of the NHS is proposed which will try to combine the efficiency principle existing in the financing of hospital services with an equity principle reflected in a ceiling on target expenditure for each district.

Economics, Hospital↗

Study of three AMPFLPs (D1S80, 3'ApoB and YNZ22) in the population of the north of Portugal.

Allele and genotype frequencies for D1S80, 3'ApoB and YNZ22 loci have been determined in a population sample of the North of Portugal using the polymerase chain reaction (PCR) amplification and nonradioactive detection. The distribution of genotypes in the three polymorphisms studied is in agreement with expected values according to the Hardy-Weinberg equilibrium. The combined chance of exclusion for the three systems is 0.96, and the combined power of discrimination is 0.99.

Alleles↗

Tuberculosis surveillance and evaluation system in Portugal.

Tuberculosis is an important public health problem in Portugal, so a new and improved computerized programme for the surveillance and evaluation of tuberculosis data was set up to obtain more precise information of relevant epidemiological indices, besides helping clinicians and regional coordinators to evaluate the local situation, such as clinical features, results of treatment and surveillance of failures and abscondings. Data stored on the computer is described in detail. The evolution of the incidence of tuberculosis is presented, together with more detailed figures for 1990. These figures were obtained with the former, cruder process. The present system is now being deployed and checked, after 1 year's trial and evaluation by some of its users. Its results will be presented in a future publication.

Databases, Factual↗

Serological survey of Leishmania infection in dogs from the municipality of Peso da Régua (Alto Douro, Portugal) using the direct agglutination test (DAT) and fast agglutination screening test (FAST).

Leishmaniasis caused by Leishmania infantum is a prevalent disease in dogs and humans. A serological survey of Leishmania infection in dogs was carried out in the endemic region of Alto Douro (north Portugal). Two hundred and ninety-four dogs from the municipality of Peso da Régua were examined for clinical signs of canine leishmaniasis (CanL), and sera samples were evaluated by the direct agglutination test (DAT) and the fast agglutination screening test (FAST). The sero-prevalence of infection was 20.4%, after screening the study population by FAST and subsequent confirmation by DAT. The overall prevalence of disease was 3.1%. Only 15.0% of the sero-positive dogs had clinical signs of CanL. A high degree of agreement (88.4%; kappa value = 0.71) was found between DAT and FAST. This study further demonstrates that FAST can be used as a simple, rapid and sensitive screening test for canine Leishmania infection in areas of high endemicity and, together with DAT, is a valuable tool in the assessment of CanL.

Agglutination Tests↗

Oxidative stress biomarkers in two resident species, mullet (Mugil cephalus) and flounder (Platichthys flesus), from a polluted site in River Douro Estuary, Portugal.

Exposure of marine animals to certain pollutants can enhance reactive oxygen species (ROS) production with subsequent damage to macromolecules and alterations in oxidant defences levels. Aimed at correlating the tissue concentration of certain contaminants (PCBs, DDT) with antioxidant defence levels and oxidative damages, two fish species with different life strategies (mullet, Mugil cephalus, and flounder, Platichthys flesus) were collected in the Douro Estuary (NW Portugal). After capture, the fish were left to depurate for 1 month in clean seawater. The levels of the two antioxidant enzyme activities revealed that they are species-dependent with mullet's livers showing higher superoxide dismutase (SOD) (13.2+/-0.5 U/mg protein) and catalase (CAT) (15.5+/-1.0 mmol/min/mg protein) activities than flounder (SOD: 7.9+/-0.9 U/mg protein; CAT: 11.1+/-0.8 mmol/min/mg protein). After 1 month in captivity the antioxidant enzymes activities in liver decreased in mullets, while for flounders the responses were not consistent because during the experimental period flounders did not ate and responses of antioxidant enzymes and oxidative damages were dependent on the fasting condition. The liver oxidative damages were evaluated by estimating oxidised lipids and proteins. Both species showed similar levels for these two parameters. The hepatic lipid peroxidation in flounder increased after 1 month in captivity, while in mullet an increase was observed only in summer and autumn. The oxidised protein content increased for both species after the depuration period. This study reveals differences between species under oxidative stress when exposed to pollutants. In a clean environment, the mullet's primary antioxidant defences decreased indicating that the animals living in Douro estuary were facing an oxidative stress. The data indicate that, namely in mullet, the presence of pollutants induce oxidative stress responses.

Animals↗

Unusually common cystic fibrosis mutation in Portugal encodes a misprocessed protein.

A561E, a novel cystic fibrosis (CF) associated mutation in the first nucleotide binding domain of CFTR, is the second most common CF mutation in Portugal. Properties of the A561E-CFTR protein were studied by immunoblotting, pulse-chase, immunocytochemistry, and MQAE halide-efflux assay in stably transfected BHK cells. Altogether, results presented here suggest that A561E causes protein mislocalization in the endoplasmic reticulum where the mutant protein must be trapped by the quality control mechanism. We conclude that A561E originates a protein trafficking defect, thus belonging to class II of CFTR mutations. As it is the case for F508del-CFTR (the most common CF mutant), low temperature treatment partially rescues a functional A561E-CFTR channel, suggesting that substitution of glutamic acid for alanine at position 561 does not completely abolish CFTR function. Pharmacological strategies previously reported for treatment of CF patients with the F508del mutation could thus be also effective in CF patients bearing the A561E mutation.

Amino Acid Sequence↗

Analysis of serpentinophytes from north-east of Portugal for trace metal accumulation--relevance to the management of mine environment.

In north-east of Portugal, the serpentinized area is about 8000 ha with a characteristic geology and flora. The serpentine plant community and respective soils were analyzed to examine the trace metal budget in different tissues of the plants exhibiting resistance to trace metals. One hundred and thirty five plant species belonging to 39 families and respective soils have been analyzed for total Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn. Substantial amounts of Ni, Cr, Co and Mn were detected in plant tissues which are listed below: NI: Alyssum serpyllifolium (38105); Bromus hordeaceus (1467); Linaria spartea (492); Plantago radicata (140); Lavandula stoechas (118) and Cistus salvifolius (114); CR: L. spartea (706.7); Ulmus procera (173.4); A. serpyllifolium (129.3); Cistus ladanifer (40.8); L. stoechas (29.5); P. radicata (27.81); Setariopsis verticillata (25.7); Plantago lanceolata (24); Digitalis purpurea (23.4); Logfia minima (23.1); Arenaria querioides (23); Hieracium peleteranum (22.7); Arenaria montana (14.5); CO: A. serpyllifolium (145.1); L. spartea (63.2); P. radicata (10.4); H. peleteranum (7.3); Lepidium heterophyllum (6.9); A. querioides (6.6); C. salvifolius (6.5); C. ladanifer (6.3); L. stoechas (6.1); Anthyllis lotoides (6.1); L. minima (6.1); Euphorbia falcata (5.7) and B. hordeaceus (5.6); MN: A. serpyllifolium (830); L. spartea (339); L. stoechas (187.1); L. minima (182.7); Castanea sativa (125); Spergula pentandra (124); P. radicata (119); Cytisus striatus (115.4); Quercus pyrenaica (110); Teucrium scorodonia (109.4); Fraxinus vulgaris (109); Anthyllis sampaiana (108); Quercus ilex (108). The significance of serpentine flora, need for conservation of these fragile and environmentally invaluable plant resources for possible use for in situ remediation of metalliferous substrates are presented in this paper.

Environment↗

Evaluation of the toxicity of two soils from Jales Mine (Portugal) using aquatic bioassays.

Soil contamination can be one path for streams and groundwater contamination. As a complement of chemical analysis and total contaminants determination, bioassays can provide information on the bioavailable fraction of chemical compounds, focusing on the retention and habitat function of soils. In this study the evaluation of the toxicity of two soils from the abandoned Jales Mine (Portugal) regarded both functions. The buffer capacity of soils was tested with bioassays carried out using the cladoceran Daphnia magna and the marine bacteria Vibrio fischeri. The habitat function of soils was evaluated with the reproduction bioassay with the collembolan Folsomia candida. The Microtox solid-phase test was performed with V. fischeri using soil as test medium, and soil elutriates were extracted to perform the Microtox basic test, and an immobilization and reproduction bioassay with D. magna. The marine bacteria showed high sensitivity to the soil with low heavy metal content (JNC soil) and to JNC soil elutriates, while the soil with highest heavy metal content (JC soil) or soil elutriates exposure did not cause any toxic effect. In the bioassays with D. magna, organisms showed sensitivity to JNC and also to JC soil elutriates. Both mobilization and reproduction features were inhibited. The bioassay with F. candida did not reflect any influence of the contaminants on their reproduction. Although JNC soil presented lower heavy metal contents, elutriates showed different patterns of contamination when compared to JC soil and elutriates, which indicates different retention and buffer capacities between soils. Results obtained in this study underlined the sensitivity and importance of soil elutriate bioassays with aquatic organisms in the evaluation strategy in soil ERA processes.

Aliivibrio fischeri↗

Sources of phenolic compounds in two catchments of southern Portugal - effect of season, land use and soil type.

Water quality monitoring in reservoirs used for human water consumption, carried out by the Alentejo Regional Authorities of the Environment (south Portugal), revealed seasonal peaks of phenolic compounds above the water-quality legislation. The main objectives of this work were to identify the main phenolic compounds present in water and soil leachates, and to determine the sources of the seasonal concentrations of phenolic compounds in two catchments with different land use patterns: Roxo and Santa Clara catchments. The main phenolic compound detected was 2,4-dinitrophenol (2,4-DNP), both in stream water and soil leachates, with concentrations higher in Roxo catchment. Roxo catchment represents a larger agricultural area than Santa Clara, and it is likely that the origin of the 2,4-DNP is associated with the use of pesticides. A peak of 2,4-DNP concentrations was observed in stream water of both catchments during February, when farmers plough their fields and apply pesticides. The 2,4-DNP peak was probably caused by a precipitation event shortly after the application of pesticides, increasing their transfer from land surfaces to adjacent streams. The leaching behaviour of 2,4-DNP was strongly dependent on the type of soil and pH. In soils with high clay content and low pH, 2,4-DNP was easily adsorbed, and its runoff from the soil to adjacent streams was reduced. Ribeira de Santa Vitória, from Roxo catchment, was the only stream showing a high abundance of vegetation, and the lowest concentrations of 2,4-DNP in water. Plants may play a role in removing contaminants from stream water.

2,4-Dinitrophenol↗

Monitoring of ambient air PCDD/F levels in Portugal.

As part of a monitoring program conducted by IDAD--Institute for Environment and Development and supported by regional municipal solid waste (MSW) management authorities, dioxin concentrations in ambient air were measured in three regions of Portugal: Porto, Lisbon and Madeira. These independent studies were performed with the intention of providing data as a basis for the evaluation of potential impacts of the operation of recently built MSW incinerators. Thus, 170 samples were collected in nine different sites from January 1999 till present. The measured levels revealed an extremely variable content of PCDDs/PCDFs depending both on the area and the season of the year. Samples taken in Porto and Lisbon reveal a similar homologue structure even if concentrations measured in the Porto region are significantly greater. Data from Madeira is characteristic of a remote site with some of the congeners concentrations below the detection limit.

Air Pollutants↗

Enzymatic biomarkers in the crab Carcinus maenas from the Minho River estuary (NW Portugal) exposed to zinc and mercury.

Zinc and mercury are common contaminants of estuaries and coastal ecosystems where they may induce adverse effects on the biota. Carcinus maenas is a key-species in several European estuaries, living in close association with the sediment where a considerable number of environmental contaminants, including zinc and mercury may accumulate. In the present study, the acute effects of zinc and mercury on C. maenas from the Minho River Estuary (NW Portugal) were investigated by using the activity of the enzymes cholinesterase (ChE), lactate dehydrogenase (LDH) and glutathione S-transferases (GST) as effects' criteria. Crabs were exposed for 96h to several concentrations of Zn(2+) (0, 1.84, 3.71, 7.39 and 14.79mg/l) or Hg(2+) (0, 0.09, 0.19, 0.37 and 0.74mg/l). The choice of Zn(2+) test concentrations was based on the LC50 value (14.86mg/l) determined in a first part of the study, while the choice of Hg(2+) concentrations was based on values from the literature. At the end of the bioassays, eye, muscle and hepatopancreas tissues were isolated and used for ChE, LDH and GST determinations, respectively. Zinc significantly inhibited ChE activity (p<0.05, EC50=14.68mg/l), caused significant alterations of GST activity (p<0.05) and induced LDH activity (p<0.05) at the exposure of 14.79mg/l. Mercury significantly inhibited ChE activity (p<0.001, LOEC=0.09mg/l, EC50=0.235mg/l) and increased both GST (p<0.05, LOEC=0.774mg/l) and LDH activities (p<0.05, LOEC=0.119mg/l). These results suggest that both metals interfere with cholinergic neurotransmission in C. maenas by inhibiting ChE activity. In addition, they also interfere with GST activity and this may reduce the capacity of detoxification of some chemicals and/or to increase the probability of oxidative stress to occur. Furthermore, both metals increase LDH activity, suggesting an interference with energy production pathways. Therefore, the presence of zinc and mercury in estuaries at concentrations in the mg/l or mug/l range, respectively, may have a negative impact on C. maenas.

Animals↗

Biochemical responses of the marine mussel Mytilus galloprovincialis to petrochemical environmental contamination along the North-western coast of Portugal.

Following the development of urban and industrial centres petrochemical products have become a widespread class of contaminants. The aim of this study was to investigate the effects of petrochemical contamination in wild populations of mussels (Mytilus galloprovincialis) along the NW Atlantic coast of Portugal by applying antioxidant and energetic metabolism parameters as biomarkers. For that, mussels were collected at five sampling sites presenting different petrochemical contamination levels. To evaluate the mussels' antioxidant status, enzymatic activities of catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, glutathione S-transferases, as well as glutathione redox status were evaluated in gills and digestive glands of mussels collected from the selected sites. Lipid peroxidation was determined in the same tissues to quantify cellular oxidative damage. Furthermore, to investigate how energetic processes may respond to these contaminants, the activity of NADP(+)-dependent isocitrate dehydrogenase was determined in mussels' digestive glands, and octopine dehydrogenase was determined in mussels' posterior adductor muscle. Furthermore, the concentrations of aliphatic hydrocarbons, unresolved complex mixture and polycyclic aromatic hydrocarbons (PAHs) were quantified in mussels' tissue, and abiotic parameters were quantified in water samples collected at each site. Several biomarkers showed statistically significant differences among sampling sites. The redundancy analysis (RDA) used to perform the integrated analysis of the data showed a clear separation of the sampling sites in three different assemblages, which are in agreement with the PAHs levels found in mussels tissues. In addition, the RDA indicated that some of the selected biomarkers may be influenced by abiotic parameters (e.g. salinity, pH, nitrates and ammonia). The approach selected for this study seems to be suitable for monitoring petrochemical contamination.

Animals↗

Spatial variation of mercury levels in nesting Bonelli's eagles from Southwest Portugal: effects of diet composition and prey contamination.

Mercury (Hg) was determined in adult Bonelli's eagles (Hieraaetus fasciatus) and their avian prey, from samples of feathers collected between 1992 and 2001 at the nesting sites of 21 pairs in Southwest Portugal. Eagle Hg levels showed great variation, reflecting primarily differences in diet composition and food chain biomagnification. Concentrations were positively correlated with the dietary proportion of insectivorous and omnivorous birds (e.g. egrets, corvids and thrushes), with very low levels for pairs feeding mainly on herbivores (e.g. rabbits, pigeons and partridges). Differences in prey contamination among breeding territories added to dietary effects in determining variation of Hg levels in eagles, shaping a spatial pattern that was largely consistent with a source of contamination in a coal-burning power-plant lying upwind of the study area. Despite this presumed contamination, Hg levels seemed to be of little concern to this eagle population, though there might be subtle deleterious effects on the reproductive output of a few pairs. This study emphasizes the need to account for dietary effects when biomonitoring Hg contamination using birds of prey.

Animals↗

Survey of organotin compounds in rivers and coastal environments in Portugal 1999-2000.

In the period from April 1999 to May 2000, organotin pollution, namely butyl and phenyltins, was investigated in coastal and continental waters (46 stations), estuarine sediments (15 stations) and mussels (Mytilus galloprovincialis) (13 stations) throughout Portugal. Sampling points were chosen in areas of specific industrial, agricultural and harbor activities. Butyltins (BTs) were the only tin species identified of which tributyltin (TBT) was found in the whole area. Concentrations of TBT in river water ranged from 3 to 30 ng L(-1) (as Sn), marine sediment ranged from 4 to 12 microg kg(-1) (as Sn), whereas concentrations in mussel tissue ranged from 2.5 to 490 microg kg(-1) (as Sn). Given that some water samples appeared to be contaminated by higher monobutyltin (MBT) and dibutyltin (DBT) concentrations, the role of biological degradation and direct inputs from agricultural and industrial applications areas are discussed. The study compares depleted butyltin pollution in sediments and mussels of the Portuguese coastline associated with antifouling paints with previously reported levels. Inputs in river waters are more related to (i) PVC leaching and (ii) industrial sources, in some cases discharged by municipal wastewaters.

Agriculture↗

Contribution of anthropogenic pollutants to the increase of tropospheric ozone levels in the Oporto Metropolitan Area, Portugal since the 19th century.

The main purpose of this study was to evaluate the contribution of anthropogenic pollutants to the increase of tropospheric ozone levels in the Oporto Metropolitan Area (Portugal) since the 19th century. The study was based on pre-industrial and recent data series, the results being analyzed according to the atmospheric chemistry. The treatment of ozone and meteorological data was performed by classical statistics and by time-series analysis. It was concluded that in the 19th century the ozone present in the troposphere was not of photochemical origin, being possible to consider the respective concentrations as reference values. For recent data a cycle of 8h for ozone concentrations could be related to traffic. Compared to the 19th century, the current concentrations were 147% higher (252% higher in May) due to the increased photochemical production associated with the increased anthropogenic emissions.

Air Pollutants↗

Ecological survey of Saccharomyces cerevisiae strains from vineyards in the Vinho Verde Region of Portugal.

One thousand six hundred and twenty yeast isolates were obtained from 54 spontaneous fermentations performed from grapes collected in 18 sampling sites of three vineyards (Vinho Verde Wine Region in northwest Portugal) during the 2001-2003 harvest seasons. All isolates were analyzed by mitochondrial DNA restriction fragment length polymorphism (mtDNA RFLP) and a pattern profile was verified for each isolate, resulting in a total of 297 different profiles, that all belonged to the species Saccharomyces cerevisiae. The strains corresponding to seventeen profiles showed a wider temporal and geographical distribution, being characterized by a generalized pattern of sporadic presence, absence and reappearance. One strain (ACP10) showed a more regional distribution with a perennial behavior. In different fermentations ACP10 was either dominant or not, showing that the final outcome of fermentation was dependent on the specific composition of the yeast community in the must. Few of the grape samples collected before harvest initiated a spontaneous fermentation, compared to the samples collected after harvest, in a time frame of about 2 weeks. The associated strains were also much more diversified: 267 patterns among 1260 isolates compared to 30 patterns among 360 isolates in the post- and pre-harvest samples, respectively. Fermenting yeast populations have never been characterized before in this region and the present work reports the presence of commercial yeast strains used by the wineries. The present study aims at the development of strategies for the preservation of biodiversity and genetic resources as a basis for further strain development.

Agriculture↗

Application of temperature gradient gel electrophoresis to the study of yeast diversity in the estuary of the Tagus river, Portugal.

Temperature gradient gel electrophoresis (TGGE) was employed for the assessment of yeast diversity in the estuary of the Tagus river (Portugal). The molecular detection of yeasts was carried out directly from water samples and, in parallel, a cultivation approach by means of an enrichment step was employed. A nested PCR was employed to obtain a fungal amplicon containing the D2 domain of the 26S rRNA gene. For identification the TGGE bands were extracted, re-amplified, and sequenced. Fourteen fungal taxa were detected and all except one were yeasts. Most yeast sequences corresponded to members of the Ascomycota and only three belonged to the Basidiomycota. Five yeasts (four ascomycetes and one basidiomycete) could not be identified to the species level due to the uniqueness of their sequences. The number of species detected after enrichment was higher than the number of taxa found using the direct detection method. This suggests that some yeast populations are present in densities that are below the detection threshold of the method. With respect to the analysis of the yeast community structure, our results indicate that the dominant populations belong to Debaryomyces hansenii, Rhodotorula mucilaginosa, Cryptococcus longus, and to an uncultured basidiomycetous yeast phylogenetically close to Cr. longus. The combined analysis of direct detection and cultivation approaches indicates a similar community structure at the two sampled sites since nine species were present at both localities.

Base Sequence↗

Allele frequencies of sixteen STRs in the population of Northern Portugal.

Allele frequencies of sixteen autossomal short tandem repeats (STRs), D3S1358, VWA, D16S539, D8S1179, D21S11, D18S51, TH01, FGA, D5S818, D13S317, D7S820, TPOX, CSF1PO, Penta D, Penta E (included in the PowerPlex 16 kit), and the SE33 (PowerPlex ES Monoplex System SE33) were determined in a sample of 200 healthy unrelated individuals from the north of Portugal.

DNA Fingerprinting↗