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Wild leporids have been gaining interest and prominence in the scientific and social community worldwide. While endangered of extinction in its native territory, the Iberian Peninsula, where it has a key role in the Mediterranean ecosystems, the European rabbit (Oryctolagus cuniculus) is considered a plague in Australia, due to the great economic and ecological consequences of its presence in the territories. The impact of viral diseases on the Leporidae family’ members, namely on the European rabbit, has been largely recognized worldwide since the early 50s, due to the emergence of myxomatosis and, from the mid-80s onwards, due to the emergence of rabbit haemorrhagic disease virus 1 and 2. More recently, in 2018, a recombinant myxoma virus emerged with the ability to infect and cause severe disease in the Iberian hare (Lepus ganatensis). Also, a new gammaherpesvirus was described in Iberian hares, associated with myxoma virus infections. In this chapter, we revise the main viral infectious treats to the native leporids of the Iberian Peninsula. The recovery of the European rabbit populations, as well as of other leporid species around the world, is currently a major challenge for the scientific and social communities and policy-makers. If we fail, the ripple effects on the trophic web will be so dramatic that are likely to be unrecoverable.

Research Areas: Cell Biology Commercial hare and rabbit immortalized cell lines are extremely limited regarding the many species within the lagomorpha order. To overcome this limitation, researchers and technicians must establish primary cell cultures derived from biopsies or embryos. Among all cell types, fibroblasts are plastic and resilient cells, highly convenient for clinical and fundamental research but also for diagnosis, particularly for viral isolation. Here, we describe a fast and cheap method to produce primary fibroblast cell cultures from leporid species, using dispase II, a protease that allows dermal–epidermal separation, followed by a simple enzymatic digestion with trypsin. This method allows for the establishment of an in vitro cell culture system with an excellent viability yield and purity level higher than 85% and enables the maintenance and even immortalization of leporid fibroblastic cells derived from tissues already differentiated. info:eu-repo/semantics/publishedVersion

Background West Nile virus (WNV) is a mosquito-borne pathogen with a globally expanding geographical range. Many European countries regularly report outbreaks affecting both human and equine populations, with an increase in epidemic activity in the past decade. Portugal, on the far west of Europe, has not yet experienced large outbreaks of WNV, despite its suitable Mediterranean climate. Objectives To provide a comprehensive, data-driven perspective on the past epidemiology, surveillance and transmission suitability of WNV in Portugal. Methods Literature and database reviews were performed, covering the period between 1966 and 2020 on existing evidence for WNV circulation. Climatic data was used to estimate the theoretical transmission potential of WNV, and to analyse long-term climate trends between 1981 and 2019. Results Existing data show that WNV-capable mosquito species are spread across the country, while WNV molecular and serological evidence is vastly concentrated in the south. Our analyses suggest that two different climate types, characteristic of the north and south of Portugal, contribute to quantitative differences in WNV theoretical transmission potential. This supports the north-south divide observed from previous molecular and serological evidence. Due to long-term changes in local climate, suitability for WNV transmission is shown to have been slowly increasing in the country. Discussion Data and theoretical analyses support the notion that the south of Portugal is more suitable for WNV transmission, while not rejecting the possibility of transmission in the north. Mosquito monitoring has been effective, but surveillance of key sentinel species remains passive, creating important knowledge gaps for epidemic control and public health. To date, evidence supports WNV circulation, but it remains unclear whether it is endemic or sporadically transmitted. This study supports a timely change towards WNV active surveillance, to avoid and mitigate potential public health emergencies as those recently observed in other European countries.

Aves polyomavirus 1 (APyV) is the causative agent of acute disease in birds and causes high mortality rates in nestlings. Infections have been reported worldwide in a significant number of caged bird species, such as parrots, caiques, budgerigars, lovebirds, and macaws. However, the number of complete viral sequences available in public databases is scarce, especially those with associated clinical histories. In this study, the clinical, pathological, epidemiological, and molecular characteristics of 2 APyV strains detected in Portugal are described. In the autumn of 2015, a 2-month-old female green-winged macaw ( Ara chloroptera ) from a small breeder died with signs of dehydration, weight loss, and depression, raising the suspicion of polyomavirus infection. Histopathological analysis revealed lesions compatible with APyV infection, and the presence of polyomavirus in several organs was confirmed by polymerase chain reaction. From the cohabitants tested ( n = 14), 1 eclectus parrot ( Eclectus roratus ), which was more than 1-year-old, was also APyV DNA-positive. The full genomic sequences of the 2 strains were obtained and found identical, suggesting a single introduction in the premises and the occurrence of subsequent infections by the same virus. When compared with sequences of other APyVs available in public databases, high nucleotide similarity percentages were obtained, confirming the close genetic relationship among polyomaviruses worldwide. Interestingly, strain APV7, detected in a white-bellied parrot in 2008 in Japan, was the closest strain to those isolated in this report. Attempts to isolate the virus in eggs and cell lines failed. Phylogenetic analysis was performed by the Bayesian method to determine the phylogenetic classification of the macaw and parrot strains. Both clustered into group V, together with other strains from different bird species with no host or spatial-temporal relationships being observed.

West Nile fever (WNF) is an emergent disease in Europe, under surveillance in the European Union. Following a 5-year period of apparent silence (autumn 2010 to summer 2015), West Nile virus (WNV) reemerged in the South of Portugal, in July 2015. Here we present data from the onset, geographic location within mainland Portugal, and outcome of clinical cases of WNV infection in horses in 2015 and 2016. During the transmission seasons of 2015 and 2016, twenty-seven horses, most symptomatic (n = 20) were found positive to IgM, pr-E immunoglobulins and VNT, leading to the subsequent report to Animal Disease Notification System of the European Commission (ADNS) by the Portuguese National Authority for Animal Health. Outbreaks occurred in the middle summer (August) and early/mid autumn (October/November) of 2015 and 2016, in the southern regions of the country (Alentejo and Algarve). Compared with the previous WNV transmission seasons of 2004 and 2010, a higher number of cases were reported in 2015 and 2016. The results of our study contribute to increase information concerning the geographic areas affected and time period for WNV transmission risk in Portugal.

Avian influenza (AI) is an infectious viral disease usually asymptomatic in wild birds that can spread to domestic poultry and cause large-scale outbreaks. Some of the AI viruses (AIV) can cross the species barrier and induce fatal disease to humans, a matter of great concern worldwide. Early detection of AIV is of major importance for disease control, since prompt implementation of adequate measures can prevent spread of the virus and therefore further outbreaks. This paper reports the development and validation of a blocking ELISA using a monoclonal antibody against a conserved structural protein for the serologic diagnosis of all AI virus subtypes from domestic bird species, allowing the quick, easily automated and low-cost screening of a high number of farms. The test will be of great value not only for surveillance, but also for monitoring the efficiency of vaccination programs. Cut-off values were established in 20% of inhibition for turkey sera and in 50% of inhibition for chicken and duck sera. Estimations of 100% specificity and 100% sensitivity were obtained based on the results of known AI positive (n=130) and negative (n=208) sera, including serum samples from birds infected with other common avian viral pathogens (n=7). ROC analysis showed an area under curve (AUC) of 1.0 for chicken, duck and turkey sera, indicating that the blocking ELISA was able to perfectly discriminate between negative and positive samples of any of the poultry species tested. Inter- and intra-assay coefficients of variation were above the acceptance threshold. Furthermore, the ELISA titers were similar to the known hemagglutination inhibition titers of three positive reference sera indicating sensitivity comparable with the golden standard HI method. The method here described is an economically attractive alternative to the commercial ELISAs currently available.

The opposition between intrinsic (I) and extrinsic (E) religious motivation was operationalised by Allport and Ross through a Religious Orientation Scale (ROS). However, empirical studies using the ROS or other derived instruments have not always been able to confirm this clear opposition. This study presents the results of the Portuguese version of the Age-Universal I/E Scale and of a Current Religious Practice Scale in four convenience community samples of the Portuguese population (global N = 932). Exploratory factor analysis enables the definition of three factors. Factor 1 groups I and Extrinsic Personal (Ep) items and is similar to the first factor obtained in studies on European non-Protestant populations. However, confirmatory analysis suggests that the I and Ep dimensions can be distinguished from each other, although they are very closely related. The difference between these two dimensions was clearer in the subgroup of practising Catholics.

É estudada a prevalência de perturbações depressivas numa amostra de mulheres com idades compreendidas entre os 35 e os 65 anos, utentes dos cuidados de saúde primários. Todas as mulheres (n = 179) responderam à versão portuguesa da Center for Epidemiologic Studies Depression Scale (CES-D) e, as que aceitaram (n = 127), participaram numa entrevista clínica estruturada adaptada do módulo de avaliação das perturbações do humor da Structured Clinical Interwiew for DSM-IV Axis I Disorders (SCID-I), a fim de avaliar a presença de várias formas de depressão segundo os critérios do DSM-IV. Verificou-se que 13% destas mulheres satisfaziam os critérios para o diagnóstico de episódio depressivo major, 19% satisfaziam os critérios para perturbação distímica e 5,5% satisfaziam os critérios propostos para episódio depressivo minor. São analisadas as relações da idade, nível de instrução, estatuto conjugal, ter filhos, ter filhos em casa, ter filhos a cargo e outras variáveis sociodemográficas com a ocorrência de perturbações depressivas. DOI: http://dx.doi.org/10.17575/rpsicol.v19i1/2.397

Porcine circovirus type 2 (PCV2) is a spherical and non-enveloped virus belonging to the genus Circovirus of the Circoviridae family with a single stranded circular DNA genome. This virus, already detected worldwide, has been associated to several diseases and was implicated as the etiological agent of a disease named postweaning multisystemic wasting syndrome. Several methods have been described for the detection of PCV2, being real-time PCR the most simple and reliable. As far as we know, all the real-time PCR systems described until now are based on ORF2 gene, that exhibit the highest variability. This paper reports the development and validation of a real-time PCR targeted to ORF1 and based on a TaqMan probe for the detection of porcine circovirus type 2 DNA in swine samples. Due to the lack of PCV1 samples, the ability of the test to discriminate between PCV1 and PCV2 positive samples was evaluated in silico. Estimations of 100% specificity and 100% sensitivity were obtained based on the qPCR results with panel of 81 swine samples (known PCV2-positive (n = 50); known PCV2-negative (n = 17); samples positive to other common swine viral pathogens (n = 13) and one sample from a BFDV-positive parrot (n = 1)). Intra- and inter-assay coefficients of variation obtained with three positive samples of different viral charges in five replicates or in five independent assays were below the acceptance threshold. The limit of detection determined with a recombinant plasmid containing the amplicon, led to conclude that this assay can detect at least three plasmid copies.

We report the detection of rabbit haemorrhagic disease virus 2 (RHDV2) in the Madeira archipelago, Portugal. Viral circulation was confirmed by RT-qPCR and vp60 sequencing. Epidemiological data revealed the outbreak initiated in October 2016 in Porto Santo affecting wild and domestic rabbits. It was then detected three months later on the island of Madeira. Five haplotypes were identified and a genetic overall similarity of 99.54 to 99.89% was observed between the two viral populations. Unique single nucleotide polymorphisms were recognised in the Madeira archipelago strains, two of which resulting in amino acid substitutions at positions 480 and 570 in the VP60 protein. Phylogenetic investigation by Maximum Likelihood showed all the vp60 sequences from the Madeira archipelago group together with high bootstraps. The analysis also showed that the Madeira archipelago strains are closely related to the strains detected in the south of mainland Portugal in 2016, suggesting a possible introduction from the mainland. The epidemiological data and high genetic similarity indicate a common source for the Porto Santo and Madeira RHDV2 outbreaks. Human activity related to hunting was most probably at the origin of the Madeira outbreak.

The genetic relationships between 10 rabbit hemorrhagic disease strains collected in Portugal between 2006 and 2013, originated in the mainland and Azorean islands, were investigated based on the vp60 gene variability. A genetic diversity ranging from 2% to 13% was determined among the 10-vp60 complete sequences revealing a significant level of genetic heterogeneity between same strains. Phylogenetic Bayesian analysis showed that the Portuguese RHDV strains fell within different genogroups, namely G1, G5 and G6. Interestingly, all strains obtained from Azores, where RHDV was first detected in 1988, belong to G5 genogroup. G5 strains, that were not identified in the continent so far, seem to be the dominant group in these Atlantic islands. G1-related strains belonging to the Iberian group 3 (n=3) and G6 (RHDVa) strains (n=2) were identified among the samples originated in mainland which were collected between 2006 and 2008. Although the presence of G1 and G6 in Portugal had been shown before, our data refines the time of circulation of these strains until at least 2008. In summary, this study revises the epidemiological information of RHDV in Portugal since it reports for the first time the presence of G5 strains in Azores and demonstrates the circulation of G1 and G6 strains in mainland Portugal until the late 2000s.

Empirical research on the relationship between menopause and depression has found contradictory results. Some authors suggest that depression is related to menopausal symptoms and not to menopausal status per se. Our aim is to study the relationship between menopausal status and depressive symptoms in a sample (n = 728) of middle-aged Portuguese women, taking into account the presence of menopausal symptoms. Depressive symptoms were measured with the Centre for Epidemiological Studies Depression Scale (CES-D). There is a relatively high significant correlation between depressive and menopausal symptomatology (rs = .47). The increase of depressive symptomatology in perimenopause is still significant, even when adjusting for the presence of menopausal symptomatology.

The line of research on middle-aged women's depression that we have been developing for over a decade has pointed to the frequency and severity of this disorder among Portuguese women, particularly less educated women. These women are more vulnerable and often receive a strictly medical response, which leaves little room for the elaboration of their complaints. We present a general overview of the results of this research line on depression in middle-aged women and the positive effect of a group intervention carried out in a health center where part of this research took place.

This report presents the results of a multiyear (2005 to 2009) study of avian influenza virus (AIV) occurrence in wild birds in Portugal. A total of 5691 samples from wild birds belonging to 13 different orders were examined. Ninety-three samples tested positive for AIV by matrix reverse transcriptase-polymerase chain reaction, giving a total prevalence of 1.63%. Twenty-one viruses were successfully cultured in embryonated chicken eggs, which represent a rate of viral infectivity of 22.6% in the samples. Nine subtypes of haemagglutinin (H1, H3 to H7, H9 to H11) and eight subtypes of neuraminidase (N1 to N4, N6 to N9) were identified in 20 different combinations. The most prevalent subtypes of haemagglutinin detected were H5, H1 and H4, while for neuraminidase subtypes N2 and N6 were the most common. The subtype combinations H4N6 and H1N1 were predominant (15.1%). All H5 and H7 viruses detected in the present study were low pathogenic for poultry as determined by the sequence of amino acids at the cleavage site of haemagglutinin. The full-length nucleotide sequences of five H5, one H7 and five N3 genes were analysed phylogenetically. The Bayesian analysis revealed that all but one of the strains analysed were closely related to isolates detected in the same period in North and Central European countries. Three H5N3 isolates, all from 2007, formed a separate cluster in both H5 and N3 phylogenetic trees. This study provides evidence that various subtypes of AIV, including subtypes H5 and H7, circulate in Portugal, which may pose a risk to industrial poultry.

The circulation of West Nile virus in Portugal was assessed by serological surveys conducted during 2004-2010 in horses and birds. The detection of WNV antibodies in both species in all the years covered by the study as well as the presence of anti-WNV IgM in symptomatic horses that had not traveled outside the country, support the notion that WNV circulates in Portugal.

Beak and feather disease virus (BFDV), a member of the genus Circovirus, was detected in six dead African grey parrots (Psittacus erithacus) in Portugal. The complete nucleotide sequences of these six BFDVs (PT05, PT08, PT08-2, PT08-3, PT09, and PT09-2) were determined and analyzed. The seven open reading frames (ORFs) described for other BFDVs were detected in all strains, except for PT05 and PT08, in which ORFs 4 and 7 are absent. Bayesian inference of phylogeny based on complete genomes of BFDVs isolated in Portugal and 32 other BFDVs found in other parts of the world revealed that PT05 is included in lineage IV, whereas the others form a new proposed genotype lineage IX. The nucleotide diversity ranged from 2% to 12% between the BFDV strains isolated in Portugal and other BFDVs found worldwide.

Vaccination programs for the control of bluetongue (BT) in ruminants have limitations due to difficulties in differentiating infected from vaccinated animals (DIVA). To overcome this problem a DIVA test that looks at a differential immune response to bluetongue virus (BTV) non-structural protein 3 (NS3) was developed. The NS3 encoding gene of strain BTV4/22045/PT04 was inserted into expression vector pET-28a and expressed in Escherichia coli strain JM109. Recombinant NS3 protein was used as an antigen in an indirect ELISA (NS3-ELISA) to measure the serologic response to NS3 protein in cattle and sheep. Following a cattle vaccination/challenge experiment with a bivalent inactivated BTV 2-4 vaccine, NS3 antibodies were detected at approximately 15 days post-infection in control unvaccinated animal, while vaccinated animals did not develop detectable NS3 antibodies and, with exception of one, remained negative even after virus challenge. The inactivated vaccine induced antibodies against the major core structural protein (VP7) of BTV as well as neutralizing antibodies in all animals. To evaluate the applicability of NS3-ELISA in field scenario, a total of 562 serum samples collected from uninfected, BTV-infected and vaccinated animals were tested for NS3 antibodies. Taken together, the results confirm that NS3 antibodies were induced to the greatest levels in animals infected with BTV in comparison to the levels induced in animals immunized with inactivated BTV vaccines, implying that antibody response to NS3 allows the differentiation between infected and vaccinated animals.

FOLLOWING on from our letter published earlier this year ( VR , January 31, 2015, vol 176, p 130), which reported the detection of rabbit haemorrhagic disease virus variant 2 (RHDV-2) in wild rabbits from four islands of the western and central group of the Azorean archipelago, we would like to communicate the diagnosis of RHDV-2 in domestic rabbits from Terceira Island. Two adult rabbits were diagnosed with RHDV-2 (Duarte and others 2015). They originated from the localities of Quatro-Ribeiras and Sao Bento, located on the north and south coasts of Terceira Island. The deaths …

Rabbit haemorrhagic disease virus 2 (RHDV2) is widespread in several countries of Western Europe, but it has not been introduced to other continents. However, between late 2014 and early 2015, the presence of RHDV2 was confirmed outside of the European continent, in the Azores, initially in the islands of Graciosa, Flores, S. Jorge and Terceira. In this study we report the subsequent detection of RHDV2 in wild rabbits from the islands of Faial, St. Maria and S. Miguel, and display the necropsy and microscopic examination data obtained, which showed lesions similar to those induced by classical strains of RHDV, with severe affection of lungs and liver. We also disclose the result of a genetic investigation carried out with RHDV2 positive samples from wild rabbits found dead in the seven islands. Partial vp60 sequences were amplified from 27 tissue samples. Nucleotide analysis showed that the Azorean strains are closely related to each other, sharing a high genetic identity (>99.15%). None of the obtained sequences were identical to any RHDV2 sequence publically known, hampering a clue for the source of the outbreaks. However, Bayesian and maximum likelihood phylogenetic analyses disclosed that Azorean strains are more closely related to a few strains from Southern Portugal than with any others presently known. In the analysed region comprising the terminal 942 nucleotides of the vp60 gene, four new single nucleotide polymorphisms (SNP) were identified. Based on the present data, these four SNPs, which are unique in the strains from Azores, may constitute putative molecular geographic markers for Azorean RHDV2 strains, if they persist in the future. One of these variations is a non-synonymous substitution that involves the replacement of one amino acid in a hypervariable region of the capsid protein.

A specific real time RT-PCR for the detection of RHDV2 was developed and validated using RHDV and RHDV2 RNA preparations from positive field samples. The system was designed to amplify a 127 nucleotide-long RNA region located within the vp60 gene, based on the alignment of six sequences originated in Portugal, obtained in our laboratory, and 11 sequences from France and Italy. The primers and probe target sequences are highly conserved in the vast majority of the RHDV2 sequences presently known. In the sequences showing variability, only one mismatch is found per strain, usually outlying the 3' end of the primer or probe hybridization sequences. The specificity of the method was demonstrated in vitro with a panel of common rabbit pathogens. Standardization was performed with RNA transcripts obtained from a recombinant plasmid harboring the target sequence. The method was able to detected nine RNA molecules with an efficiency of 99.4% and a R(2) value of 1. Repeatability and reproducibility of the method were very high, with coefficients of variation lower than 2.40%. The assay was proven a valuable tool to diagnose most of RDVH2 circulating strains, and may be also useful to monitor viral loads, and consequently, disease progression and vaccination efficacy.

RABBIT haemorrhagic disease virus (RHDV) causes a highly infectious and lethal disease in European rabbits ( Oryctolagus cuniculus ). In Portugal the disease was first reported on the island of Madeira in 1987 and, in the following year, in the Azores. The first report of the disease on the mainland occurred in 1989. Since then, the disease has become endemic throughout the country. The genetically distinct RHDV variant 2 (RHDV-2) was reported on the Portuguese mainland …

The complete genomic sequence of a sheep lentivirus isolate that presents a slow/low phenotype in vitro has been determined. The virus, designated P1OLV, was isolated from lung cells of a naturally infected sheep in Portugal. Three overlapping DNA fragments amplified by PCR, and encompassing the entire viral genome were cloned and sequenced. This isolate has an overall similarity of approximately 80% with the K1514 Maedi Visna virus (MVV) and approximately 70% with the caprine arthritis encephalitis virus (CAEV) Co strain. Phylogenetic analysis based on SU and RT nucleotide sequences grouped P1OLV with previously reported ovine MVV. To determine the virus replication rate, sheep choroid plexus (SCP) and lung cells, macrophages (MØ), and goat synovial membrane (GSM) cells were inoculated with either P1OLV or with the lytic North American strain WLC-1. Viral RNA in culture supernatants was measured by one-tube real time quantitative RT-PCR. Significant differences were observed between the replication rates of the two viruses, with WLC-1 growing rapidly and to high levels in all the cells tested, while P1OLV replicated more slowly and to lower levels inducing persistent infections in lung and SCP cells. The U3 region of the LTR of P1OLV lacks the sequence repeats that are present in the LTRs of WLC-1 and MVV prototype K1514 and that contain additional binding sites for the AML(vis) transcriptional factor. To evaluate the contribution of LTR in the virus replication rate in vitro, we measured the basal activity of the promoter from P1OLV and WLC-1 in a luciferase-driven gene expression assay and lower levels of expression were achieved for P1OLV. The genetic and biological properties of P1OLV will be useful for the study of virus transcriptional factors and genes that may be responsible for the slow/low phenotype.

Summary: The Center for Epidemiologic Studies Depression Scale (CES-D) is a depression scale designed to measure depressive symptomatology in the general population. The authors developed a Portuguese version of this scale. This article presents the results of the three studies carried out with this version. These results support the reliability and the validity of the scale. The CES-D proved to be sensitive to variations in the intensity of depressive symptomatology and can be used in epidemiological studies or as a screening instrument in a clinical setting. Sex does have some influence on the values of sensitivity and specificity but the educational level has a greater influence on scores: Subjects with a lower education level tend to obtain higher scores with the same level of depressive symptomatology as evaluated by the clinical interview.

Members of the family Adenoviridae are divided into five genera and infect a wide variety of vertebrates with a narrow host range, usually restricted to one species. Due to the high genetic diversity and distinct genomic organization, classification of adenoviruses is difficult to achieve and often performed by phylogenetic analysis. The most commonly used region for phylogenetic inference of adenoviruses is the DNA polymerase (AdPol) gene carried out at amino acid level. In this paper we investigated the suitability of the U exon to discriminate adenoviruses. The tree based on this genus-common feature, obtained with 23 short amino acid sequences, offered a clearest discrimination of the members of the adenovirus family (Adenoviridae) than the trees generated with the complete or partial polymerase protein sequences. Therefore, our results demonstrate that the U exon is an effective tool for a refined phylogenetic inference and genus classification of the Adenoviridae family.

A monoclonal antibody (MAb) blocking ELISA (Blck-ELISA) was developed to detect antibodies against Maedi-Visna virus (MVV) in sheep sera. The assay employs a MAb directed against the envelope protein p90 of the virus in a sandwich blocking procedure. To determine whether the MAb was a potential antibody for developing a Blck-ELISA, a collection of three hundred sera obtained from several sheep flocks known to be infected with MVV were used to examine the sensitivity of the Blck-ELISA. A total of 50 serum samples originating from a flock free of MVV were tested to assess the specificity of the assay. The results were compared with a commercial indirect ELISA (I-ELISA) and samples giving a conflicting or doubtful result were tested by immunoblot. The Blck-ELISA proved to be specific, sensitive and it showed high reproducibility and low variability.

This paper describes the history and current status of professional psychology in Portugal where a unique perspective combines training, research, and practical contributions from Europe and the Americas with their own history of psychological tradition and expertise. Training in professional psychology includes Social Psychology and Educational and Vocational Guidance specializations in addition to Clinical Psychology and Psychotherapy and Counseling for the professional degree, Licenciatura. Advanced degrees are offered in Environmental Psychology, Career Development, Social Cognition, and other areas, primarily for academic positions. Research in all of these areas is expected to have applied outcomes that contribute to individual well being and an improved quality of life for the entire population. The result has been a rapid development of an indigenous professional psychology to address mental health, social, and environmental concerns that compel psychological attention and resources worldwide as well as those problems of local and national origins.

Pain complaints and depressive symptoms seem to be risk factors for future depressive episodes. We present a one-year follow-up study on Portuguese women and primary health care (initial sample: n = 503). We used the Center for Epidemiologic Studies Depression Scale (CES-D), a short questionnaire on pain, and clinical interviews. We used forward stepwise logistic regression analysis to define models that enable the prediction of developing a major depressive episode in the one-year follow-up. We concluded that nondepressed women with (a) high scores (≥28) at the CES-D, (b) a high severity of pain index, and (c) fatigue complaints at the baseline were particularly at risk.

West Nile virus (WNV) and Japanese encephalitis virus (JEV) are important mosquito-borne viruses of the Flaviviridae family, associated with encephalitis, mainly in humans and horses. WNV is also pathogen for many bird species. The incidence of human and animal WNV infections in Europe has risen, mostly in recent years, and JEV was detected in 2011 in mosquitoes collected in Italy and may emerge in Europe in the same way as other flaviviruses had emerged recently (USUTU and Bagaza virus) and should be regarded as a potential threat to public health. Prompt identification and discrimination between WNV and JEV provides critical epidemiological data for prevalence studies and public and animal health management policies. Here we describe a quantitative one-step duplex TaqMan RT-PCR, targeting non-structural protein 2A gene (NS2A-qRT-PCR), based on only one primer pair and two probes for differential diagnosis of WNV and JEV. Also this assay enables the detection of both WNV lineages (WNV-1 and WNV-2). To access the specificity of NS2A-qRT-PCR a panel of different arboviruses were used. The assay was shown to be specific for both WNV lineages (WNV-1 and WNV-2), WNV related Kunjin virus and JEV, since no cross-reactions were observed with other tested arboviruses. Sensitivity of the assay was determined using serial dilutions of in vitro-transcribed RNA from WNV and JEV. The duplex NS2A-qRT-PCR assay was shown to be very sensitive, being able to detect 10 copies of WNV and JEV RNA. This assay is a suitable tool for the diagnosis of WNV and JEV, and provides a valuable addition to the methods currently available for routine diagnosis of these zoonoses and for surveillance studies.

The myxoma virus (MYXV) causes severe infections in European rabbits that may reach mortality rates up to 100% depending on the viral strain. The typical symptoms and lesions induced by the virus are usually enough to permit the correct clinical diagnosis. However, in peracute forms the infection may be accompanied by unspecific symptoms. Sudden death may also occur without evident clinical signs of myxomatosis. Likewise, a clinical diagnosis of atypical forms of myxomatosis (amyxomatous) is often complicated and delayed due to the scarceness of skin lesions. As the disease control often depends on an early and unequivocal diagnosis of MYXV, laboratorial methods play a relevant role in the confirmation of MYXV infection. This study describes the development and validation of a novel, high accurate real time polymerase chain reaction assay (rtPCR) for the detection of MYXV. Primers were designed to amplify a 125-bp within the gene M000.5L/R, which is duplicated in the termini of the genome and is unique among Leporipoxvirus. The assay was negative for SFV and other poxviruses and was able to detect 2.6 copies of MYXV DNA proving the effectiveness, specificity and sensitivity of this diagnosis tool. The rtPCR has been applied successfully in INIAV laboratory for routine diagnosis of myxomatosis since 2005.

The exposure of wild carnivores to viral pathogens, with emphasis on parvovirus (CPV/FPLV), was assessed based on the molecular screening of tissue samples from 128 hunted or accidentally road-killed animals collected in Portugal from 2008 to 2011, including Egyptian mongoose (Herpestes ichneumon, n = 99), red fox (Vulpes vulpes, n = 19), stone marten (Martes foina, n = 3), common genet (Genetta genetta, n = 3) and Eurasian badger (Meles meles, n = 4). A high prevalence of parvovirus DNA (63%) was detected among all surveyed species, particularly in mongooses (58%) and red foxes (79%), along with the presence of CPV/FPLV circulating antibodies that were identified in 90% of a subset of parvovirus-DNA positive samples. Most specimens were extensively autolysed, restricting macro and microscopic investigations for lesion evaluation. Whenever possible to examine, signs of active disease were not present, supporting the hypothesis that the parvovirus vp2 gene fragments detected by real-time PCR possibly correspond to viral DNA reminiscent from previous infections. The molecular characterization of viruses, based on the analysis of the complete or partial sequence of the vp2 gene, allowed typifying three viral strains of mongoose and four red fox’s as feline panleukopenia virus (FPLV) and one stone marten’s as newCPV-2b type. The genetic similarity found between the FPLV viruses from free-ranging and captive wild species originated in Portugal and publicly available comparable sequences, suggests a closer genetic relatedness among FPLV circulating in Portugal. Although the clinical and epidemiological significance of infection could not be established, this study evidences that exposure of sympatric wild carnivores to parvovirus is common and geographically widespread, potentially carrying a risk to susceptible populations at the wildlife-domestic interface and to threatened species, such as the wildcat (Felis silvestris) and the critically endangered Iberian lynx (Lynx pardinus). published

The frequency of changes experienced by menopausal women is variable in different cultures. It is our purpose to identify the symptoms that Portuguese women consider to be related to menopause. We also wish to evaluate the relations between these symptoms and menopausal status, sociodemographic and lifestyle factors, and medication—menopausal hormone therapy (MHT) and psychotropics. In women with a low educational level, the frequency of symptoms and the consumption of psychotropic medication is higher, and the practice of exercise is lower, as is the use of MHT. Our results may be useful in providing guidance to health care professionals.

Porcine circovirus type 2 (PCV2) belongs to the genus Circovirus of the Circoviridae family and has been associated with a disease named postweaning multisystemic wasting syndrome (PMWS). In Portugal the disease caused severe problems mainly between 1995 and 2003, but is nowadays less evident probably as the result of vaccination programmes initiated in 2007. This study reports the molecular analysis of 22 PCV2 strains obtained from domestic pigs originated from different regions of Portugal. One hundred and seventy-seven samples from PCV2 suspected cases received between 2003 and 2010 were analysed, 79 (44.6%) of which tested PCV2 PCR-positive. The entire genomes of 22 viruses were amplified and sequenced. Two major groups were observed, according to the genome length and in single nucleotide polymorphisms. The nucleotide similarity found among the PCV2 strains ranged between 94% and 99%. The Bayesian inference of phylogeny based on the genome of the strains studied revealed the existence of two distinct genetic groups consistent with the nucleotide sequence observations. The majority of the strains branched in genotype PCV2b, while six strains with a common geographical origin obtained between 2007 and 2009, were included in genotype PCV2a. The analysis performed with cap and rep genes confirmed that cap is a reliable alternative to full-length genome based phylogenetic studies of PCV2.

Este estudo descreve a sintomatologia relacionada com a menopausa nos períodos pré‑menopausa, menopausa e pós‑menopausa, em relação com a sintomatologia depressiva, nível educacional, estatuto conjugal e outras variáveis sociodemográficas, numa amostra de 127 mulheres com idades compreendidas entre os 35 e os 65 anos, que compareceram a uma consulta com o médico de família num Centro de Saúde dos arredores de Lisboa. As mulheres responderam a um questionário com questões relativas ao estatuto percebido de menopausa e à presença de sintomas com ela relacionados, e participaram numa entrevista clínica estruturada adaptada do módulo de avaliação das perturbações do humor da Structured Clinical Interwiew for DSM‑IV Axis I Disorders (SCID‑I). Descreve‑se a frequência da sintomatologia relacionada com a menopausa e analisa‑se a sua variação em relação com o nível educacional e outras variáveis sociodemográficas. A relação entre o estatuto percebido de menopausa e a sintomatologia depressiva é também analisada, tomando em consideração o nível educacional. This study is based on the data collected in an investigation regarding the Primary Health Care patients. The results relating to a sample of 127 women with ages ranging between 35 and 65 years of age are analyzed. All these women answered a Questionnaire concerning the perceived menopause status and the menopause symptomatology and participated in a structured clinical interview, in which the presence of various forms of depression according to the DSM‑IV criteria was analyzed. They also answered a questionnaire concerning the perceived menopause status as well as symptoms or alterations related with menopause. The frequency of the symptomatology related with menopause is studied and its variation in relation to the level of education, and other sociodemographic variables is analyzed. The relationship between the perceived menopause status and the depressive symptomatology is also studied, taking into consideration the level of education.

Apresentação do número temático. DOI: http://dx.doi.org/10.17575/rpsicol.v19i1/2.395