The moderating roles of nostalgia activators, culture, and individual characteristics

Santini, F. D. O., Lim, W. M., Ladeira, W. J., Pinto, D. C., Herter, M. M., & Rasul, T. (2023). A meta‐analysis on the psychological and behavioral consequences of nostalgia: The moderating roles of nostalgia activators, culture, and individual characteristics. Psychology and Marketing, 40(10), 1899-1912. https://doi.org/10.1002/mar.21872While previous research suggests that nostalgia can stimulate future-oriented motivation and goal pursuit, the presence of conflicting findings complicate our understanding of the psychological and behavioral consequences of nostalgia. This study introduces and empirically validates a theoretical framework for nostalgia through a meta-analysis of 90 effect sizes and 9757 aggregate samples from 22 experimental studies. Our findings enrich understanding of the effects of nostalgia, showing that compared to a control, nostalgia enhances consumers' sense of pleasantness, self-continuity, attitudes, and behavioral intentions. We also identify several moderators that shape the effects of nostalgia, namely nostalgia activators, culture, and individual characteristics. Intriguingly, stronger behavioral intentions manifest in response to nostalgic events (rather than objects), realistic (over nonrealistic) scenarios, visual (as opposed to textual) priming, and female-majority (vs. male-majority) samples. However, we observe that long-term (vs. short-term) oriented cultures exhibit diminished nostalgia effects. Furthermore, we find no significant variations in responses based on age (older vs. younger), type of nostalgic experience (personal vs. collective), and sample type (students vs. nonstudents). Collectively, our meta-analysis reveals the intricate dynamics of nostalgia, emphasizing its profound impact on consumer behavior as shaped by moderators such as nostalgia activators, culture, and individual characteristics, thereby providing a multifaceted and nuanced understanding of nostalgia.publishersversionpublishersversionpublishe

Experimental feeding of Sergentomyia minuta on reptiles and mammals: comparison with Phlebotomus papatasi

Background: Sergentomyiaminuta (Diptera: Phlebotominae) is an abundant sand fly species in the Mediterranean basin and a proven vector of reptile parasite Leishmania (Sauroleishmania) tarentolae. Although it feeds preferentially on reptiles, blood meal analyses and detection of Leishmania (Leishmania) infantum DNA in wild-caught S.minuta suggest that occasional feeding may occur on mammals, including humans. Therefore, it is currently suspected as a potential vector of human pathogens. Methods: A recently established S.minuta colony was allowed to feed on three reptile species (i.e. lizard Podarcissiculus and geckos Tarentolamauritanica and Hemidactylusturcicus) and three mammal species (i.e. mouse, rabbit and human). Sand fly mortality and fecundity were studied in blood-fed females, and the results were compared with Phlebotomuspapatasi, vector of Leishmania (L.) major. Blood meal volumes were measured by haemoglobinometry. Results: Sergentomyiaminuta fed readily on three reptile species tested, neglected the mouse and the rabbit but took a blood meal on human. However, the percentage of females engorged on human volunteer was low in cage (3%) and feeding on human blood resulted in extended defecation times, higher post-feeding mortality and lower fecundity. The average volumes of blood ingested by females fed on human and gecko were 0.97 µl and 1.02 µl, respectively. Phlebotomuspapatasi females readily fed on mouse, rabbit and human volunteer; a lower percentage of females (23%) took blood meal on the T.mauritanica gecko; reptilian blood increased mortality post-feeding but did not affect P.papatasi fecundity. Conclusions: Anthropophilic behaviour of S.minuta was experimentally demonstrated; although sand fly females prefer reptiles as hosts, they were attracted to the human volunteer and took a relatively high volume of blood. Their feeding times were longer than in sand fly species regularly feeding on mammals and their physiological parameters suggest that S.minuta is not adapted well for digestion of mammalian blood. Nevertheless, the ability to bite humans highlights the necessity of further studies on S.minuta vector competence to elucidate its potential role in circulation of Leishmania and phleboviruses pathogenic to humans. Graphical abstract: [Figure not available: see fulltext.].publishersversionpublishe

Caracterização de vírus específicos de insetos: análise da sua diversidade genética e relação com outros vírus

Os arbovírus são responsáveis por doenças com impacto significativo na saúde humana, sendo transmitidos a humanos e outros animais por insetos e outros vetores invertebrados. Entre estes últimos, os mosquitos representam um dos mais importantes vetores conhecidos por servirem de vetores a arbovírus patogénicos para os humanos, de que são exemplos os vírus da dengue e Zika. Por muito tempo, a pesquisa de vírus foi impulsionada pelo impacto que estes agentes impõem à saúde humana/animal/plantas, mas os desenvolvimentos nas últimas décadas nos domínios das tecnologias de sequenciação de alto rendimento e bioinformática permitiram melhorias nas estratégias de descoberta de vírus, o que, por sua vez, levou a um aumento no número de vírus peculiares que vieram a ser descobertos em rastreios virológicos, alguns com replicação restrita em células de vertebrados. Acredita-se que esses vírus, designados vírus específicos de insetos, tenham impacto nulo ou baixo na saúde animal e, provavelmente por isso mesmo, tenham permanecido à sombra de arbovírus patogénicos. No entanto, nas últimas décadas eles tornaram-se no foco da nossa atenção, não apenas pela sua extensa diversidade e estratégias incomuns de replicação restrita nalguns hospedeiros, mas também pelo seu potencial de interferir na replicação de arbovírus. Desde então, diversos vírus específicos de insetos foram descobertos em várias famílias de vírus, com vírus específicos de mosquitos associados especialmente às famílias Flaviviridae, Mesoniviridae e Parvoviridae. Neste projeto procurou-se detetar e analisar novas sequências de vírus específicos de insetos dessas três famílias virais em mosquitos coletados em Portugal, Angola e Moçambique. A diversidade genética, reconstrução filogenética e avaliações filodinâmicas foram então executadas, usando tanto sequências genómicas geradas de novo, bem como de sequências disponíveis em bases de dados públicas. Novas sequências de flavivírus específicos de insetos ditos "clássicos" (cISF) foram detetadas em pools de mosquitos das três regiões geográficas, e diferentes sub-linhagens de cISF foram caracterizadas. A sua análise filodinâmica sugeriu que a dispersão de cISF no espaço e no tempo deverá ser recente e bastante dinâmica. Por outro lado, embora dados insuficientes não tenham permitido uma análise filodinâmica completa com base em sequências de mesonivírus, foi realizada uma extensa revisão taxonómica, que incluiu a análise de sequências semelhantes a mesonivírus (meson-like viruses) recentemente detetadas em outros organismos que não mosquitos. Por fim, também procurámos analisar entre os parvovírus de invertebrados aqueles que têm sido incluídos no género Brevihamaparvovirus, cuja distribuição até ao momento parece ser restrita a algumas espécies de mosquitos. Os seus genomas parecem evoluir sob forte seleção negativa e também são caracterizados por baixa entropia, tal como foi igualmente observado para flavivírus e mesonivírus. Também realizámos uma revisão taxonómica do táxon (o primeiro para brevihamaparvovírus) e efetuámos a sua primeira reconstrução filodinâmica.Arboviruses are responsible for impactful viral diseases and are transmitted to humans and other animals by insect and other arthropod vectors. Among the latter, mosquitoes pose as one of the most important hematophagous vectors known to carry pathogenic arboviruses for humans, like dengue and Zika. For a long while, virus research was driven by the negative impact viruses impose on human/animal/plant health, but recent developments in next-generation sequencing and bioinformatics have allowed for improvements on virus discovery strategies. In turn, these have led to an increase in the number of peculiar viruses detected in viral screening-based studies, some of which display restricted replication in vertebrate cells. These viruses, designated insect-specific viruses, are thought to have none-to-low impact on animal health and have endured in the shadow of pathogenic arboviruses. However, in the last decades they have become the focus of our attention, not only due to their extensive diversity and unusual host-restriction strategies, but also because of their potential to interfere with the replication of arboviruses. Insect-specific viruses have since been discovered in multiple virus families, with mosquito-specific viruses especially associated with the Flaviviridae, Mesoniviridae and Parvoviridae families. In this project we sought to detect and analyze new insect-specific virus sequences from these three viral families in mosquitoes collected in Portugal, Angola and Mozambique. Genetic diversity, phylogenetic reconstruction and phylodynamic assessments were then executed, using both new generated sequences and sequences available in public databases. New classical insect-specific flavivirus (cISF) sequences were detected in mosquito pools from these three geographic regions, and different sub-lineages inside the cISF cluster were characterized. Phylodynamics analyses suggested that cISF dispersion over space and time could be recent and quite dynamic. On the other hand, while insufficient data did not allow for a full phylodynamic analysis based on mesonivirus sequences, an extensive taxonomy revision was performed, that also included the analysis of sequences similar to mesoniviruses (meson-like viruses) recently detected in organisms other than mosquitoes. Finally, we also sought to analyze among the parvoviruses of invertebrates those included in the Brevihamaparvovirus genus, that have been restricted (so far) to a few mosquito species. Their genomes seem to evolve under strong purifying selection and are also characterized by low entropy, as also observed for flaviviruses and mesoniviruses. We also performed a taxonomic revision of the taxon (the first ever for brevihamaparvoviruses), and attempted their first ever phylodynamic reconstruction

An experiment on primary education in Angola

How should parents be mobilized for education in Africa? We implemented a large-scale field experiment in Angolan primary schools, including three treatments: an information campaign at home, simple parents’ meetings at school, and a combination of both. Our measures of parental mobilization include beneficial practices at home, contacts with teachers, and participation in school institutions. We find that the information increased parents’ involvement at home but had no impact on engagement at school, while the meetings had the opposite effects. After mobilizing parents, only the combined treatment improved management practices and facilities in schools, teachers’ attitudes, and parents’ satisfaction.preprintinpres

a forced-choice identification study with Hungarian native speakers

UIDB/03213/2020 UIDP/03213/2020 2020.05740.BDAuditory perceptual experiments with Portuguese L2 are scarce, especially within the scope of the European variety (EP). In this study, we aim at observing the assimilation of the EP vowels [ɐ] and [ɨ] by Hungarian native speakers, and their perceptual learning after some contact with Portuguese. A multiple forced-choice identification experiment was run with two groups of Hungarian speakers – a group with and a group without previous contact with EP. The results show that the categorization fell into the closest phonetic categories of the L1. The results also indicate a learning effect for [ɨ], with a recategorization path from [y] to [ø]. As for [ɐ], no major differences were observed between groups. This result is consistent with difficulties observed in the classroom environment, where [ɐ] is particularly challenging to Hungarian learners.authorsversioninpres

Nise da Silveira and the revolutionary use of art1 as Occupational Therapy in the psychiatric context

UIDB/04666/2020 UIDP/04666/2020Este artículo propone una revisión de la biografía y la obra de Nise da Silveira, psiquiatra brasileña que realizó una labor revolucionaria en el tratamiento de las personas con enfermedades mentales. Sigue una metodología cualitativa con base narrativa y documental donde se destaca la importancia de su acción innovadora, que rescata la Terapia Ocupacional con herramientas artísticas en las que se privilegia la libre expresión y el contacto afectivo como alternativa a los tratamientos convencionales de la época, como el electrochoque y la lobotomía. Se relacionan, desde una perspectiva comparativa, conceptos de la psicología analítica de Jung –como los mitos, arquetipos y el inconsciente colectivo– y la experiencia terapéutica psiquiátrica de Nise. La presentación de casos prácticos de algunos pacientes permite concluir que los símbolos y arquetipos revelados en las imágenes artísticas creadas en la Terapia Ocupacional tienen un impacto estructurante y curativo que se sigue, hoy en día, en diversos contextos terapéuticos. This paper aims to review the biography and work of Nise da Silveira, a Brazilian psychiatrist who carried out revolutionary work in the treatment of the mentally ill. This paper follows a qualitative methodology with a narrative and documentary base, highlighting the importance of her innovative action that rescued Occupational Therapy with artistic tools, in which free expression and affective contact are favored as an alternative to the conventional treatments of the time, such as electroshock and lobotomy. From a comparative perspective, Jung’s analytical psychology concepts such as myths, archetypes and the collective unconscious are related to Nise’s psychiatric therapeuti experience. The presentation of some patients case studies leads to the conclusion that the symbols and archetypes revealed in the artistic images created in occupational therapy have a structuring and healing impact, which is still followed today in various therapeutic contexts.publishersversionpublishe

Dynamics of HIV-1 transmission in Europe: a guidance for evidence-based prevention strategies

Introdução: Para controlar a pandemia de VIH a UNAIDS desenvolveu os objetivos 95- 95-95 para serem atingidos até 2030. A concretização destes objetivos pode ser dificultada pelo aparecimento de mutações de resistência devido ao uso intensivo da terapia antirretroviral ou também à existência de indivíduos com apresentação tardia (IAT) ao diagnóstico. Estes IAT não só impactam os resultados dos seus próprios tratamentos como são também uma ameaça para alcançar os objetivos da UNAIDS, uma vez que podem potenciar, de forma inconsciente, a transmissão do VIH. Objetivos: Primeiro, identificar as caraterísticas sociodemográficas e clínicas dos indivíduos infetados com VIH-1, bem como identificar os determinantes da apresentação tardia em Portugal e na Europa. Segundo, descrever os padrões de resistência transmitida (TDR) e de resistência adquirida (ADR) em indivíduos infetados com VIH-1 seguidos na Europa, comparar os seus padrões de resistência IAT e indivíduos com apresentação nãotardia (IANT) e analisar as mutações de resistência aos antirretrovirais nos diferentes subtipos de VIH-1. Para finalizar, descrever e caraterizar os clusters de transmissão (CT) de VIH-1 na Europa e comparar o papel dos IAT com os IANT nos CT do VIH-1. Metodologia: No primeiro estudo, a base de dados utilizada incluiu dados clínicos e sociodemográficos de indivíduos infetados com VIH-1 seguidos no Hospital Egas Moniz, Centro Hospitalar de Lisboa Ocidental (CHLO), Lisboa, Portugal, entre1984 e 2017. Nos outros estudos, utilizou-se a EuResist Integrated Database (EIDB) que inclui dados sociodemográficos, clínicos e genómicos de indivíduos infetados com VIH-1 seguidos na Europa (Portugal, Espanha, Alemanha, Luxemburgo, Rússia, Reino Unido e Itália) entre 1981 e 2019. Para a análise dos CTs, foram utilizadas informações de indivíduos infetados pelos subtipos mais prevalentes (B, A e G). Resultados: No primeiro estudo, 68,7% dos indivíduos infetados com VIH-1 eram homens com uma mediana de idade de 37 anos (IQR 30–47). 50,6% destes indivíduos tinham apresentação tardia (AT) e desses 61,9% tinham apresentação tardia com doença avançada. Os determinantes associados à AT foram idade ao diagnóstico superior a 30 anos e origem em países da África subsaariana. No segundo estudo, entre os indivíduos incluídos na análise a mediana de idade foi igual a 33 anos (IQR: 27,0–41,0) e 74,4% eram homens. 50,4% destes indivíduos tinham AT e os determinantes associados foram idade acima de 56 anos, heterossexuais, indivíduos com origem em países africanos e com carga viral abaixo de 4,1cópias/mL. No terceiro estudo, a mediana de idades obtida foi igual a 37 anos (IQR: 27,0–45,0) e 72,2% eram homens. 71,9% dos indivíduos tinham sido infetados pelo subtipo B e 54,8% foram classificados com AT. Para AT e apresentação não-tardia (ANT) a prevalência de TDR foi 12,3% e 12,6% respetivamente, e a de ADR foi de 69,9% e de 68,2% respetivamente. As mutações mais prevalentes observadas em IAT e IANT foram K103N/S, T215rev, T215FY, M184I/V, M41I/L, M46I/L e L90M. No quarto estudo, o subtipo mais prevalente nos indivíduos infetados com VIH-1 foi o subtipo B (84,7%) seguido do subtipo G (9,4%) e subtipo A (5,9%). A idade mediana foi de 33 anos (IQR: 26,0-41,0) e 75,5% eram homens. 51,4% dos x indivíduos infetados com VIH-1 tinham AT e 21,6% estavam dentro de CTs. As análises filogenéticas demonstraram que apenas 17,6% dos IAT estavam dentro de CTs comparados com 20,2% dos IANT. Para os subtipos A e B, verificou-se que os IAT dentro de CTs foram caracterizados por uma menor percentagem de homens e por uma maior percentagem de indivíduos mais velhos comparativamente aos IANT. Para os subtipos B e G, os IAT dentro de CTs apresentaram maior percentagem de tratados comparativamente com os IANT. No subtipo G, os IAT dentro de CTs, eram maioritariamente utilizadores de drogas intravenosas comparativamente com os IANT. Analisando o tamanho dos CTs, verificou-se que os IANT pertenciam a grandes CTs (>8 indivíduos) comparativamente aos IAT. Conclusão: A AT é considerada um dos grandes obstáculos para travar a epidemia do VIH e uma ameaça à transmissão do mesmo. Os nossos resultados apresentam as características sociodemográficas e clínicas dos IAT na Europa e indicam que estes não contribuem, de forma significativa, para a transmissão dos VIH-1. Os resultados encontrados podem contribuir para o desenvolvimento de medidas de prevenção e para uma melhor compreensão das mutações de resistência e falhas terapêuticas nesta população de indivíduos.Background: To control the HIV pandemic, the UNAIDS set the 95-95-95 targets to be reached by 2030. These targets can be more difficult to achieve, whether due to the appearance of drug resistance mutations regarding the increasing use of antiretroviral therapy (ART) or due to individuals who present late at diagnosis (late presenters-LP). These individuals can not only impact treatment outcomes, but also threat UNAIDS goals, as well as potentiate the spread of HIV. Aims: First, to identify clinical and sociodemographic characteristics of HIV-1 infected patients, as well as to identify determinants of late presentation in Portugal and in Europe. Second, to describe the patterns of transmitted drug resistance (TDR) and acquired drug resistance (ADR) in HIV-1 infected patients followed in Europe (Portugal, Spain, Germany, Luxembourg, United Kingdom, Russia and Italy), to compare its patterns in late presenters (LP) vs non-late presenters (NLP), and to analyze the most prevalent drug resistance mutations among HIV-1 subtypes. And finally, to describe and characterize HIV-1 transmission clusters in Europe and to compare the role of LP vs NLP populations on HIV-1 transmission clusters (TC). Methods: For the first study, the database included clinical and sociodemographic information from HIV-1-infected patients followed in Hospital Egas Moniz, Centro Hospitalar de Lisboa Ocidental (CHLO), Lisbon, Portugal, between 1984 and 2017. For the other studies, the EuResist Integrated Database (EIDB) included socio-demographic, clinical, and genomic information from HIV-1 infected patients followed between 1981 and 2019. For the analysis of TC, information from patients infected with the most prevalent subtypes B, A and G was analyzed. Results: In the first study, 68.7% of patients were males and the median age was 37 years (IQR 30–47). 50.6% patients were LP and, of those, 61.9% were late presenters with advanced disease (LPAD). The determinants associated with LP were age at diagnosis higher than 30 years and origin from sub-Saharan Africa. In the second study, among the HIV-1 infected patients included in the analysis, the median age was 33 (IQR: 27.0–41.0) years and 74.4% were males. 50.4% were late presenters and the determinants associated with late presentation were older patients (>56), heterosexuals, patients originated from Africa and patients presenting with log VL >4.1. In the third study, the median age of HIV-1 infected individuals was 37 (IQR: 27.0–45.0) years old and 72.6% were males. 71.9% of patients were infected by subtype B and 54.8% of patients were classified as LP. For LP and NLP, the TDR prevalence was 12.3% and 12.6%, respectively, while ADR, was 69.9% and 68.2%, respectively. The most prevalent TDR drug resistance mutations, in both LP and NLP, were K103N/S, T215rev, T215FY, M184I/V, M41I/L, M46I/L, and L90M. In the fourth study, the most prevalent subtype among those infected with HIV-1 was subtype B (84.7%), followed by subtype G (9.4%) and subtype A (5.9%). The median age was 33 (IQR: 26.0-41.0) years old and 75.5% of patients were males. 51.4% of patients were classified as LP and 21.6% of patients were inside TCs. xii Phylogenetic analyses showed that only 17.6% of LPs were inside clusters compared to 20.2% of NLPs. For subtypes A and B, we found that LP inside clusters were less frequently males and were older than NLPs. For subtypes B and G, LP inside clusters were more frequently treated than NLP. In subtype G, LP inside clusters more frequently had IDU transmission route than NLP. Finally, when analyzing cluster size, we found that NLP more frequently belonged to large clusters (>8 patients) when compared to LP. Conclusion: Late presentation is a major obstacle to halt the HIV epidemic and could be a threat to HIV-1 transmission. Our results characterize the socio-demographic and clinical characteristics of LPs in Europe and, all together, indicate that LPs are not important contributors to forward HIV-1 transmission. These results help to direct prevention measures for this population and to better understand drug resistance mutations and therapeutic failure in this population of patients

Leptospira spp. in Portugal: Epidemiological and molecular study in geographic areas with different urbanization levels

A leptospirose é uma doença zoonótica infecciosa de importância global. Os roedores são os reservatórios mais comuns na disseminação de Leptospira spp.. Quando excretadas pela urina, as espiroquetas podem sobreviver por várias semanas ou meses, em condições favoráveis (solo húmido, água com pH neutro ou ligeiramente alcalino). Assim, a água e o solo contaminados tornam-se importantes veículos de transmissão para os humanos, os quais podem ser infetados pela penetração destes agentes infecciosos nas membranas mucosas intactas (nariz, boca, olhos), ou através da pele lesionada. Este estudo teve como objetivo avaliar a presença de Leptospira spp. em colecções de água doce, solos e roedores de três distritos de Portugal (Lisboa, Setúbal e Viseu). Foram recolhidas amostras de coleções de água doce distintas (superfície de lagos, lagoas, rios, riachos e fontes públicas) (ninverno=198; nverão=147), de solos (perto de coleções de água doce, latas de lixo ou áreas de vegetação) (n=104), em 15 cidades dos distritos de Lisboa, Setúbal e Viseu. Nos locais de colheita, foram avaliados alguns parâmetros químicos e físicos (por exemplo, pH, nitritos, temperatura), e registada a localização. Paralelamente às amostras ambientais, foram capturados roedores (N=35) na mesma região, para a análise molecular do sangue, urina e órgãos (rins, fígado, baço e pulmões), e cultura do tecido renal. Após a extração do DNA de todas as amostras, foram usados dois protocolos de nested-PCR com diferentes primers. Inicialmente, um protocolo de nested-PCR com primers universais do gene rrs (16S), para a detecção de Leptospira spp.. Cada amostra com amplificação de DNA de Leptospira spp. foi submetida a um segundo protocolo de nested-PCR, com primers específicos direcionados ao gene lipL32, para a deteção de espécies patogénicas de Leptospira. Um protocolo de PCR em tempo real usando primers “LipL32” foi também otimizado para a deteção de DNA de leptospiras patogénicas em amostras ambientais e de roedores (sangue total e baço), bem como para analisar a viabilidade das leptospiras em águas e solos. O protocolo de nested-PCR com primers universais permitiu a deteção de DNA leptospírico em 170 amostras de água (ninverno=116/198, 59%; nverão= 54/147, 37%) e em 88 amostras de solo (85%). Uma avaliação posterior com os primers "LipL32", permitiu a deteção de DNA de leptospiras patogénicas em 87 amostras de água (ninverno= 54/116, 47%; nverão= 33/54, 61%), em 11 amostras de solo (13%), e em 29 roedores capturados (83%). Os resultados da sequenciação demonstraram a presença de Leptospira interrogans e Leptospira borgpetersenii nas amostras ambientais, na urina e rins de roedores. Nenhum crescimento foi detetado nas culturas. Os testes de viabilidade realizados em certas amostras ambientais com DNA de leptospiras patogénicas, demonstraram que 44% (n= 4/9) continham leptospiras viáveis capazes de causar infeção em humanos.A amplificação de DNA leptospírico em mais da metade das amostras ambientais analisadas, e a evidência de que a maioria dos roedores capturados estão colonizados com leptospiras patogénicas, levanta uma importante evidência de saúde pública acerca do risco de transmissão para a população destas áreas geográficas.Leptospirosis is an infectious zoonotic disease of worldwide importance. Rodents are the most common reservoirs in Leptospira spp. dissemination. Spirochetes can survive for several weeks or months under favorable conditions (moist soil, water with neutral or slightly alkaline pH), when excreted in urine. Thus, contaminated water and soils become essential vehicles of transmission to humans, which can be infected by penetration of these infectious agents into intact mucous membranes (nose, mouth, eyes), or injured skin. This study aimed to evaluate the presence of Leptospira spp., freshwater collections, soils, and rodents from three districts of Portugal (Lisboa, Setúbal, and Viseu). Samples from distinct freshwater collections (surface of lakes, ponds, rivers, streams, and public fountains) (nwinter=198; nsummer=147), as well as soils (near freshwater collections, trashcans, or vegetation areas) (n=104), were collected in 15 cities from Lisboa, Setúbal and Viseu districts. Certain chemical and physical parameters (e.g., pH, nitrites, temperature) were evaluated at the collection sites, and the location was recorded. In parallel with environmental samples, rodents were trapped (N=35) in the same region for the molecular analysis of the blood, urine, and organs (kidneys, liver, spleen and lungs), and the culture of the kidneys macerate. After DNA extraction from all samples, two nested-PCR protocols with different primers were used. At first, a nested-PCR protocol with universal primers from the rrs (16S) gene for Leptospira spp. detection. Each sample with Leptospira spp. DNA amplification was submitted to a second nested-PCR protocol, with specific primers targeting lipL32 gene, for detection of pathogenic species of Leptospira. A real-time PCR protocol using “LipL32” primers was also optimized detecting pathogenic leptospiral DNA in environmental and rodent samples (whole blood and spleen), and to analyze leptospires viability in freshwaters and soils. The nested-PCR protocol using universal primers allowed the detection of leptospiral DNA in 170 water samples (nwinter=116/198, 59%; nsummer=54/147, 37%) and in 88 soil samples (85%). Further evaluation with the “LipL32” primers allowed the detection of pathogenic leptospiral DNA in 87 water samples (nwinter=54/116, 47%; nsummer=33/54, 61%), in 11 soil samples (13%), and in 29 rodents trapped (83%). Sequencing results demonstrated the presence of Leptospira interrogans and Leptospira borgpetersenii in the environmental samples, as well as in the rodent’s urine and kidneys. No culture growth was detected. The viability tests performed on certain environmental samples with pathogenic leptospiral DNA demonstrated that 44% (n=4/9) contained viable leptospires capable of causing human infection. The amplification of leptospiral DNA in more than half of the environmental samples analyzed, together with the evidence that most of the rodents trapped are colonized with pathogenic leptospires, raises a significant public health concern about the risk of transmission to the population from these geographic areas

Blanket Data Retention Struck Down at Last by the Portuguese Constitutional Court

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