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Herd-level seroprevalence of fasciolosis in cattle in north central Portugal.

An epidemiological study of Fasciola hepatica in cattle was implemented in the north central region of Portugal. Both an enzyme-linked immunosorbent assay and an egg shedding quantification technique were used in the follow-up of seven herds. Two of these herds were negative and the other five were positive for F. hepatica. A herd cut-off of value of 0.425 optical density was calculated and herd sensitivity (HSe) and herd specificity (HSp) were defined. Three seroprevalence studies were also implemented in the region with stratification by county sub-regions for a period of 18 months. Overall mean herd prevalence in Vagos of 11, 23 and 48% was progressively found for the three studies, respectively.

Animals↗

Molecular and phylogenetic characterization of Theileria spp. parasites in autochthonous bovines (Mirandesa breed) in Portugal.

A survey was conducted during the months of April-June 2003 in the northeast Portugal (Bragança district) in order to characterize the hemoparasite population of an autochthonous Mirandesa breed of Bos taurus. The polymerase chain reaction (PCR) analysis of the bovine blood revealed that 3 out of 116 animals were infected with Theileria and/or Babesia parasites, while reverse line blot hybridisation (RLB) analysis showed that these animals were infected with Theileria buffeli/orientalis. Cloning and sequencing confirmed the RLB results. Database sequence searches combined with phylogenetic analysis of the partial 18S ribosomal RNA gene sequences obtained enabled us to place the parasites in question as members of the T. buffeli/orientalis group, confirming the PCR/RLB diagnosis.

Animals↗

Isolation of Besnoitia besnoiti from infected cattle in Portugal.

Besnoitia besnoiti, an obligate intracellular protozoan parasite belonging to the phylum apicomplexa, is the causative agent of bovine besnoitiosis. Besnoitiosis is responsible for significant losses in the cattle industry of Africa and Mediterranean countries due to the high morbidity rate, abortion and infertility in males. The acute stage of disease is associated with the proliferative forms (tachyzoites) and is characterized by fever, whimpery, general weakness and swelling of the superficial lymph nodes. During the following chronic stage, a huge number of cysts are formed mainly in the subcutaneous tissues. This process is non-reversible, and chronic besnoitiosis is characterized by hyper-sclerodermia, hyperkeratosis, alopecia and, in bulls, atrophy, sclerosis and focal necrosis that cause irreversible lesions in the testis. In this paper we report on the identification of large cysts in the skin of a cow and a bull in Portugal, which presented loss of hair and enlargement and pachydermis all over the body. The observation of a two-layered cyst wall within the host cell, the encapsulation of the host cell by a large outer cyst wall, and the subcutaneous localization of the cysts within the host, were characteristic for B. besnoiti. The parasites were isolated from the infected animals and successfully propagated in Vero cells without prior passages in laboratory animals. Morphological characterization of B. besnoiti tachyzoites and the amplification of the 149 bp segment from the internal transcribed spacer 1 (ITS1), aided with specific primers, confirmed the identification of B. besnoiti.

Animals↗

Effect of salinity and inorganic nitrogen concentrations on nitrification and denitrification rates in intertidal sediments and rocky biofilms of the Douro River estuary, Portugal.

The regulatory effects of salinity and inorganic nitrogen compounds on nitrification and denitrification were studied in intertidal sandy sediments and rocky biofilms in the Douro River estuary, Portugal, over a 12-month period. Nitrification and denitrification rates were measured in slurries of field samples and enrichment experiments using the difluoromethane and the acetylene inhibition techniques, respectively. Salinity did not regulate denitrification in either environment, suggesting that halotolerant bacteria dominated the denitrifier communities. However, nitrification rates were stimulated when salinity increased from 0 to 15 practical salinity units. NO3- addition experiments revealed that NO3- availability stimulates denitrification rates in sandy sediments, but not in rocky biofilms; however, in rocky biofilms a positive and linear relationship was observed between denitrification rates and water column NO3- concentrations (r=0.92) during the monthly surveys. The N2O:N2 ratios increased rapidly when NO3- increased from 63 to 363 microM; however, results from monthly surveys showed that environmental parameters other than NO3- availability may be important in controlling the variation in N2O production via denitrification. Ammonium additions to sandy sediments stimulated nitrification rates by 35% for the 20 microM NH4+ addition, but NH4+ appeared to inhibit nitrification at high concentration addition (200 microM NH4+). In contrast, rocky biofilm nitrification was stimulated by 65% when 200 microM NH4+ was added.

Bacteria, Aerobic↗

Monitoring of priority pesticides and other organic pollutants in river water from portugal by gas chromatography-mass spectrometry and liquid chromatography-atmospheric pressure chemical ionization mass spectrometry.

Gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC-APCI-MS) were optimized and applied for the trace-level determination of 42 priority pesticides and 33 priority organic pollutants from European Union Directive EC 76/464. First, off-line solid-phase extraction of 200 ml of river water using an OASIS solid-phase extraction cartridge, followed by GC-MS was used. Next, selected samples that were positive to GC-MS were analyzed by LC-APCI-MS in order to detect further polar byproducts or to improve the determination of previously detected polar analytes. The transformation products of triazine pesticides like deethylatrazine (DEA) and deisopropylatrazine (DIA) and compounds such as diuron and several chlorophenols were positively identified by LC-APCI-MS. The present methodology has also been used for searching for new analytes not included in the EC 76/464 list, like Irgarol, DEA and DIA. In addition it was applied to target pollutants in 43 river water samples from Portugal during a pilot survey from April to July 1999. Atrazine followed by simazine and 2,4,6-trichlorophenol were the most ubiquitous compounds detected in this area. The levels detected of the different compounds were in the range of: 0.01-2.73 microg/l, 0.05-0.74 microg/l, 0.02-1.65 microg/l, 0.02-5.43 microg/l, 0.01-0.40 microg/l, 0.01-0.26 microg/l, 0.02-0.61 microg/l, 0.01-3.90 microg/l, 0.01-1.24 microg/l, 0.02-2.3 microg/l, 0.01-0.13 microg/l and 0.01-0.5 microg/l for atrazine, simazine, terbuthylazine, alachlor, metolachlor, Irgarol, propanil; tributhylphosphate, diuron, 2,4,6-trichlorophenol, deisopropylatrazine and deethylatrazine, respectively.

Atmospheric Pressure↗

First confirmation of human diarrhoeic poisonings by okadaic acid esters after ingestion of razor clams (Solen marginatus) and green crabs (Carcinus maenas) in Aveiro lagoon, Portugal and detection of okadaic acid esters in phytoplankton.

A new outbreak of human diarrhoeic poisonings (DSP) with esters of okadaic acid (OA) was confirmed after ingestion of razor clams (Solen marginatus) harvested at Aveiro lagoon (NW Portugal) in the summer of 2001. Accumulation of marine toxins in second order consumers was investigated in the edible parts of a shellfish predator abundant at Aveiro lagoon, the green crab Carcinus maenas. Okadaic acid was found, also in a predominant esterified form. Levels in edible parts (comprising mainly viscera) surpassed 16microg/100g. We suggest that one patient may have developed profuse diarrhoea after ingestion of a large number of green crabs contaminated with okadaic acid esters. At least 32microg OA/100g were found in a remaining sample of its meal. Domoic acid was also found but under the allowable level in force in USA of 30microg/g crab viscera. In cooked crabs, significant losses of domoic acid were found and it is not suspected to have contributed to the poisoning event, although being a vector for this toxin. The low percentage of free okadaic acid found is in accordance with a predation predominantly on benthonic shellfish (razor clams, clams and common cockle) rather than on rock mussels. These last ones present usually higher percentages of free okadaic acid. Okadaic acid was confirmed with full-scan mass spectra either in plankton and mussel extracts. Okadaic acid esters were also found in plankton extracts. Percentages between 40-60% of esterified OA were found in samples freshly extracted. Ester's percentage diminished drastically if after sonication the extract was kept at room temperature. The major part of the esters was water-soluble.

Adult↗

Spatial and temporal evolution of the levels of tritium in the Tagus River in its passage through Caceres (Spain) and the Alentejo (Portugal).

This work is the result of a collaboration between Spanish and Portuguese laboratories. The specific objective was to quantify the time evolution during 1994, 1995 and 1996 of the radioecological impact of the liquid releases of 3H from the Almaraz Nuclear Power Plant (ANPP) in the section of the Tagus River corresponding to Cáceres province in Spain and the Alentejo region in Portugal. We found that the temporal evolution of the levels of tritium depends on the management of the water held in the cooling reservoir of the ANPP and the presence of the dams that exist along the river. This management regime has a 12-month period. Also the movement of the mass of tritiated water (HTO) downriver was much faster during 1996 than 1995 or 1994 due to the hydrological differences between those years and consequently to the different amounts of water transferred between the reservoirs of the dams. From the hypothesis that hydrodynamically it is impossible to differentiate tritiated water from non-tritiated water, a model was constructed that satisfactorily reproduces the temporal evolution of the 3H in the zone of the Tagus River in which the exchange of water takes place, with the cooling reservoir of the Almaraz Nuclear Power Plant.

Fresh Water↗

Cyanobacteria diversity and toxicity in a wastewater treatment plant (Portugal).

Cyanobacteria are common in eutrophic natural waters. Being favoured by warm, stable and nutrient-enriched waters they may constitute an important part of the phytoplankton community in Wastewater Treatment Plants (WWTP). The phytoplankton communities of two ponds (facultative and maturation) of the WWTP of Esmoriz (North Portugal) were studied, with particular importance given to cyanobacteria. Mouse bioassays were performed with cyanobacteria samples during some of the blooms and ELISA assays specific for hepatotoxic microcystins were carried out. During the study period (January-July 1999) cyanobacteria were frequently dominant in the ponds ranging from 15.2 to 99.8% of the total phytoplankton density. The main species were Planktothrix mougeotii, Microcystis aeruginosa and Pseudanabaena mucicola. Mouse bioassays were performed during Oscillatoria bloom period but the results were negative, in spite of the high cyanobacteria biomass. ELISA assays were performed for both ponds but only in the maturation pond positive values were found. Microcystin concentrations (as MCYST-LR equivalents) varied from 2.3 to 56.0 micrograms/l on the margin of the pond and between 1.7 and 4.6 micrograms/l in the outflow of this pond. These values indicate that WWTP may be a source of contamination of water bodies with cyanobacteria toxins.

Animals↗

Atmospheric impact assessment and monitoring of dioxin emissions of municipal solid waste incinerators in Portugal.

In Portugal two municipal solid waste (MSW) incinerators are under construction in the metropolitan areas of Lisbon and Porto. During the development of the Environmental Impact Assessment (EIA) studies of these units the authors found several limitations and uncertainties associated to the assessment of the environmental impact. Uncertainties were particularly felt on the assessment of the environmental impacts associated with the atmospheric emissions of PCDD and PCDF. At present an External Monitoring Plan is under design.

Air Pollution↗

Volatile organic compounds in rural atmospheres of central Portugal.

Atmospheric concentrations of volatile organic compounds were measured at two rural sites in central Portugal. The sites were chosen to be in line with the summer northwesterly sea breezes in order to study the evolution of the chemical composition of air masses during transport to inland areas. The most abundant non-oxygenated hydrocarbon in the ambient air was isoprene and the monoterpenes alpha-pinene, beta-pinene and 1,8 cineol. The maximum isoprene levels (6-7 ppb) were recorded at the most inland site, suggesting an enrichment of coastal air masses with biogenic emissions during transport over eucalyptus forests. Formaldehyde was the most prominent carbonyl compound in the atmosphere but acetaldehyde and acrolein were also abundant. Concentrations of carbonyl compounds had a tendency to be higher inland, particularly for glyoxal, methyl glyoxal, methyl vinyl ketone, metacrolein and pentanal. The observed increases indicate that carbonyls were produced by photochemical oxidation of biogenic hydrocarbons in aged air masses with coastal origin. Isoprene, monoterpenes and various carbonyls exhibited pronounced diurnal variations, which are explained on the basis of emissions from vegetation, oxidation pathways of biogenic hydrocarbons and meteorological conditions.

Agriculture↗

Short-term environmental impact of clam dredging in coastal waters (south of Portugal): chemical disturbance and subsequent recovery of seabed.

The physical and chemical changes in sediment and near bottom water caused by clam dredging were examined during July and September 1999, at two locations Vilamoura (VL) and Armona (AR), south coast of Portugal. Sediment cores and near bottom water were collected simultaneously before dredging (control samples) and within short time intervals (min-h) after dredging. After dredging operations, microphytobenthos coming from the path were accumulated in the re-worked sediment (ridge). Chlorophyll a in superficial sediment increased from 1.2 microg x g(-1) before dredging to 1.7 microg x g(-1) after dredging and these higher values remained for a few hours. However, the expected increase of chlorophyll a in near bottom water due to re-suspension was not observed. After sediment disturbance an instantaneous sorption of phosphorus onto iron oxides occurred in the upper sediment layers (from 2 to 3 micromol x g(-1) before dredging to 4-5 micromol x g(-1) after dredging). A microcosm experiment showed that after sediment disturbance HPO(4)(2-) dissolved in pore water decreased from 40 to 10 microM being simultaneously sorbed onto iron oxides formed in the top layer of sediment. The ammonium, nitrates, organic nitrogen, phosphate and silicate dissolved in pore water decreased immediately after dredging activity and simultaneously an increase in near bottom water was sporadically observed. Generally, the re-establishment of seabed was reached within a short time (min-h), at both stations (VL and AR).

Animals↗

Plant community tolerant to trace elements growing on the degraded soils of São Domingos mine in the south east of Portugal: environmental implications.

The selection of trace element tolerant species is a key factor to the success of remediation of degraded mine soils. Mining activities generate a large amount of waste rocks and tailings, which get deposited at the surface. The degraded soils, the waste rocks and tailings are often very unstable and will become sources of pollution. The direct effects will be the loss of cultivated land, forest or grazing land, and the overall loss of production. The indirect effects will include air and water pollution and siltation of rivers. These will eventually lead to the loss of biodiversity, amenity and economic wealth. Restoration of a vegetation cover can fulfil the objectives of stabilization, pollution control, visual improvement and removal of threats to human beings. Thus, remediation of mine spoils/tailings and biogeochemical prospecting would rely on the appropriate selection of plant species. Plant community responds differently on their ability to uptake or exclude a variety of metals. In this work, plant species were sampled from all populations established in an abandoned copper mine of São Domingos, SE Portugal. Plants belonging to 24 species, 16 genera and 13 families were collected from the degraded copper mine of São Domingos. Plant samples were analysed for total Ag, As, Cu, Ni, Pb, and Zn. The highest concentrations of metals in soils dry matter were 11217.5 mg Pb kg(-1), 1829 mg Cu kg(-1), 1291 mg As kg(-1), 713.7 mg Zn kg(-1), 84.6 mg Cr kg(-1), 54.3 mg Co kg(-1), 52.9 mg Ni kg(-1) and 16.6 mg Ag kg(-1). With respect to plants, the higher concentrations of Pb and As were recorded in the semi-aquatic species Juncus conglomeratus with 84.8 and 23.5 mg kg(-1) dry weight (DW), Juncus efusus with 22.4 and 8.5 mg kg(-1) DW, and Scirpus holoschoenus with 51.7 and 8.0 mg kg(-1) DW, respectively. Thymus mastichina also showed high content of As in the aboveground parts, 13.6 mg kg(-1) DW. Overall, the results indicate accumulation of various metals by different plant species, with some of these metals being partitioned to the shoots. Environmental implications of these observations are discussed.

Copper↗

Burn patients in Portugal: analysis of 14,797 cases during 1993-1999.

The incidence and cause of burns in Portugal are unknown. The present study seeks to collect these data as a basis for future studies in prevention and development of treatment regimes. The data used in this work was obtained from the registries existing in The Direcção Geral de Saúde (General Health Administration) and relates to burn patients admitted to 91 Portuguese hospitals--that serve all the territory--during the period of 1993-1999. All patients selected had at least one diagnosis of burn (ICD-9: 94.###) among all the diagnoses motivating an admission. A total of 14,797 burn patients were obtained. Among all the patients admitted to hospitals, 8731 (59.0%) were male and 6066 (41.0%) were female, with a male/female ratio of 1.44:1. The mean length of in-hospital stay was 15.5+/-21.0 days. Throughout this period, 553 (3.7%) patients died in Portuguese hospitals as a result of burn injury. There were no significant differences in the number of deaths in each year, P=0.45, in contrast to the decrease of burn mortality rates reported in other studies. It is likely that the reason for this situation is the lack of investment in the last years in this area and the delay in opening new Burn Centres.

Adolescent↗

STR data (AmpFlSTR profiler plus) from north Portugal.

Allele frequencies for the nine STRs included in the AmpFlSTR Profiler Plus kit (D3S1358, VWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820) were estimated from a sample of 365-427 unrelated individuals born in north Portugal.

Alleles↗

Hereditary ataxias and spastic paraplegias: methodological aspects of a prevalence study in Portugal.

A project for studying the prevalence of hereditary ataxias (HA) and familial spastic paraplegias (FSP) in Portugal was set up in 1993. The ascertainment of patients in previous prevalence studies relied mainly on the information of hospital admissions and out-patient contacts with the neurology and other related departments at central hospitals covering the whole region surveyed. Many patients might be overlooked if large populations were studied using this method, since registers at central hospitals are very incomplete and for most part not yet computerized. On the other hand HA and FSP are rare diseases appearing in family clusters, and it would be unreasonable to undertake a sample survey based upon a suitable frame of the Portuguese population. Therefore we decided to carry out a two-phase prevalence survey at district level, involving the collaboration of all physicians working in the district health institutions and the population, in the screening of eligible subjects in phase 1. All subjects screened as positive were examined by a neurologist in phase 2. This method provided a direct estimate of false positives and false negatives were all patients also examined in phase 2, who came to our knowledge using other sources of information. The prevalence of hereditary ataxias and spastic paraplegias in the pilot district was 6.4 per 100,000 inhabitants. The sensitivity of the screening procedure was 81.2% and the predictive value of a positive screening was 25%. Considering the geographically circumscribed district nature of the populations to be studied, the comprehensive sources of case identification used and the high adherence of the health professionals involved, we believe that this method can be widely used, particularly in countries with similar health care services.

Adult↗

Notes on a plant parasite fungus in Portugal: Gymnosporangium cornutum.

A rust fungus identified as Gymnosporangium cornutum was found on Sorbus aucuparia in Serra da Estrela (Manteigas), and the disease was severe at that location. Despite the abundance and worldwide occurrence of the genus Gymnosporangium, studies in Portugal are still limited.

Basidiomycota↗

Characterisation and classification of phylloplane yeasts from Portugal related to the genus Taphrina and description of five novel Lalaria species.

Taphrina Fries is a genus of dimorphic ascomycetes comprising more than 90 species distinguished by the specific infections they produce on different vascular plants. Their filamentous states are restricted to parasitised plant tissue whereas the yeast states are saprobic and can be grown on artificial media. The latter coincide with the anamorphic phases and have been given separate nomenclatural status by the erection of the genus Lalaria R.T. Moore. In its original circumscription Lalaria included only 23 yeast states of known species of Taphrina and its creation was then redundant. Here we describe five novel species in the genus Lalaria to accommodate a total of 44 yeast isolates obtained mainly from leaf surfaces (phylloplane) of different plants in Portugal: Lalaria arrabidae sp. nov. (one strain), L. carpini sp. nov. (one strain), L. inositophila sp. nov. (37 strains), L. kurtzmanii sp. nov. (one strain) and L. veronaerambellii sp. nov. (four strains). L. inositophila was notable for its widespread occurrence since it was recovered during two consecutive years from the leaves of miscellaneous plant species. In the absence of sexual states and of unequivocal associations to particular host plants, the taxonomic relationship of the novel species to the yeast states of Taphrina available from culture collections was verified by the comparative analysis of physiological and molecular characteristics. The latter included PCR fingerprinting using single primers for microsatellite regions, sequencing of the 5' end of the 26S rRNA (LSU) gene (D1/D2 domains) and of the ITS1 and ITS2 rDNA spacer regions, and DNA-DNA hybridisation experiments. An emended description of the genus Lalaria is provided.

Ascomycota↗