Repositório RCAAP

The access to safe drinking-water is essential to assure health. It is a basic human right and a component of effective policy for health protection. Due to the increase levels of environmental pollution in some systems, it is crucial to assess water quality applying an integrative approach, combining environmental and human risk assessments, attending to surface sources pressure and the consequent drinking water security. Water treatment plants (WTPs) represent an important instrument to promote the quality of drinking water. Disinfection of water system is an essential strategy to protect human health from pathogens and prevent their regrowth during water distribution, but the reaction of disinfectant agents with organic matter can lead to the formation of disinfection by-products (DBPs). Given their widespread occurrence, potential human health impacts and (eco)toxicity associated with exposure to DBPs are of particular interest due to their potential carcinogenicity and vary non-carcinogenic effects, such as endocrine disruption. The work developed in the frame of this thesis had the ultimate aim to improve water safety plans and produce new knowledge towards a more comprehensive risk assessment, considering the occurrence and toxicity of target DBPs. In chapter I, a general description of the subject is made, considering aspects such as water treatment, the DBPs formation, main analytical methodologies to measure these compounds in drinking water, and their reported toxicity. In chapter II, with the aim of better characterizing the EPAL- Empresa Portuguesa das Águas Livres, S.A water supply system, we assessed the water quality parameters, correlating several microbiological and chemical parameters, some regulated under Portuguese /UE legal framework and others still not regulated but with environmental and human health relevance, during a 6-year period (2014-2019). For that purpose, two conventional WTPs with a technology largely implemented in many different regions, were assessed by an integrative approach as a proxy to improve processes understanding, treatment efficiency and global water quality. The outcomes of this first study contributed to the establishment of a framework to evaluate the quality of the water supply system, identifying relevant occurrence relationships, and providing meaningful information that decision makers can use for more effective water safety plans. In chapter III, we aim to characterize EPAL water supply system attending to some target UR-DBPs and given the limited validated analytical methods to detect and quantify UR DBPs, a new multi-residue gas chromatography coupled with mass spectrometry methodologies for the detection and quantification of 15 UR-DBPs in drinking water matrices was validated. The target compounds included DBPs from 4 chemically different classes, namely nitrosamines, haloketones, aldehydes and alcohols. The selected sample preparation methods included solid phase extraction (SPE), applied for nitrosamines group, and solid phase micro extraction (SPME), for the remaining DBPs. The developed analytical methods were applied to drinking water samples from different points of EPAL water supply system, including two conventional WTPs, geographically different, with distinct production capacities, periodically collected during a 4-month period. From the 15 UR-DBPs analyzed, 8 of them were detected in the analyzed drinking water samples, being the most representative DBPs belonging to aldehydes class. To the best of our knowledge, this study represents the first insights on the measurement of some of the target DBPs in drinking water matrices, considering one of the biggest Portuguese public distribution systems. In chapter IV, we review the recent literature on the effects of DBPs present in water for human consumption, mainly focusing in unregulated compounds and the putative underlying mode of action (MoA), linking the available data with adverse health outcomes. The main knowledge gaps in this field were identified, and future research priorities discussed. In chapter V, in order to improve hazard and risk assessment, 7 of the most prevalent UR DBPs were selected to be included in a toxicological approach. In this study, the developmental toxicity of the chemically-different UR-DBPs was evaluated using zebrafish (Danio rerio) embryo bioassay as a proxy animal model, and considering morphological abnormalities, % mortality and behavior endpoints. We demonstrate here toxicological effects on D. rerio development, at environmentally relevant levels, of 7 UR-DBPs. To gain additional insights into the biological functions and potential pathways disrupted by the exposure to 2 of the most toxic DBPs studied, 2-EH and NDMA, a comprehensive transcriptome assembly was also produced, and an exploratory analysis of the transcriptomic profiles were made. Before this work, no information was available regarding % of occurrence and toxicological data for some of the target DBPs analyzed. At the national context, it represents the first insights regarding the presence and characterization of target UR-DBPs, considering one of the largest Portuguese public water supply systems. In addition, an integrative analysis of the main WTPs of EPAL was made, contributing to more effective water safety plans.

African trypanosomiasis is an infectious disease caused by unicellular and extracellular parasites of the Trypanosomatidae family, including Trypanosoma brucei. Both Human and animal African trypanosomiasis are characterized by a pronounced loss of fat and skeletal muscle mass. T. brucei occupies both intravascular and extravascular compartments of the mammalian host. In the mouse model of infection, T. brucei colonizes the adipose tissue in disproportionately high numbers when compared with most other organs. The properties of the adipose tissue microenvironment, immunological and others, which enable such extensive colonization, are still incompletely understood. Moreover, the impact of adipose tissue colonization on the loss of fat mass observed during natural infections, and the impact of fat mass loss itself on disease pathology are unknown. Here, we show that during infection the adipose tissue acquires a marked transcriptomic immune signature. This correlated with the accumulation of both innate and adaptive immune cells consistent with a T helper (Th) 1 response. Moreover, as observed for the serum, we found increasingly higher titers of IgM and IgG specific for the parasite’s surface variant glycoprotein in the adipose tissue’s interstitial fluid. Similar to the blood, parasite clearance from the adipose tissue was dependent on T and B cells and the hallmark Th1 cytokine interferon gamma. Interestingly, while the complement system was dispensable to eliminate circulating parasites, mice lacking the complement component 3 showed reduced elimination of parasites from the adipose tissue. This suggests that the complement system may play an important role in the clearance of highly parasitized extravascular compartments. In addition to studying the immunological surveillance of the adipose tissue, we also studied how adipocyte metabolism is modulated by a T. brucei infection. Here, we show that during infection, white adipose tissue depots progressively reduce in size and that this reduction correlates with loss of adipocyte lipid droplet volume (i.e. fat containing organelle). In turn, this loss of adipose tissue weight is preceded by an increase in ex vivo adipocyte lipolysis. This increase in lipolysis proved to be central in loss of fat mass, as adipocyte-specific adipose tissue triglyceride lipase deficient (AdipoqCre/+-Atglfl/fl) mice, which have reduced lipolysis, were more resistant to loss of total fat mass and kept higher adipocyte lipid droplet volumes than wild-type (WT) littermate controls (Atglfl/fl). Moreover, adipocyte lipolysis showed host-protective effects, as AdipoqCre/+- Atglfl/fl mice succumbed earlier to infection than Atglfl/fl controls. In addition to increased survival, our data suggest that increased lipolysis improves local parasite burden control. On one hand, AdipoqCre/+-Atglfl/fl mice present transiently higher adipose tissue parasite burdens than Atglfl/fl controls. On the other hand, chemical induction of adipocyte lipolysis in an in vitro adipocyte-T. brucei co-culture system significantly reduces parasite growth. Lastly, we shed light on the upstream signals that induce adipocyte lipolysis. Here, we found that T. brucei promotes lipolysis in in vitro grown co-cultured adipocytes. In addition to parasite cues, we found that T and B cells are central in the induction of fat mass loss and that serum factor produce by these cells are able to induce adipocyte lipolysis. Overall, in this work we show that during a T. brucei infection a trypanocydal immune response is mounted in the adipose tissue and that as infection progresses a loss of fat mass occurs through host-protective ATGL-dependent adipocyte lipolysis

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O cancro é uma das principais causas de morte em todo o mundo, com o número de mortes a aumentar à medida que a população mundial aumenta e envelhece. No entanto, tem havido progresso na luta contra esta doença. Um dos caminhos tem sido o melhor entendimento da patogénese e base molecular do cancro, com a identificação de biomarcadores que são usados em diferentes cenários clínicos - estimar o risco de desenvolver cancro, rastreio, ajudar no diagnóstico diferencial, determinação de prognóstico, ou prever resposta à terapia - e que ajudam a melhorar a sobrevivência. Os tumores das glândulas salivares são raros. Dada a sua raridade, os clínicos pouco frequentemente lidam com este tipo de neoplasias e a maioria são tratados em centros especializados. Estes tumores têm um bom prognóstico quando estão limitados à glândula salivar. No entanto, quando o tratamento “standard” cirurgia ± radioterapia adjuvante não é possível – se o tumor é inoperável, há doença loco-regional ou à distância – as opções terapêuticas são muito limitadas e o prognóstico é reservado. O adenoma e o adenocarcinoma de células basais, com a sua morfologia basalóide característica (i.e., constituídos por células de citoplasma escasso e núcleo uniforme redondo a oval), são dois dos tumores salivares menos frequentes. A sua origem exacta é desconhecida e só recentemente foram descritas alterações moleculares recorrentes, com a identificada de mutações no gene CTNNB1 em adenomas. Neste trabalho descrevemos a experiência de 50 anos duma Instituição, um hospital terciário oncológico, em lidar com adenomas e adenocarcinomas de células basais. Na nossa série, os adenomas tiveram um óptimo prognóstico. Os adenocarcinomas, por seu lado, embora normalmente tidos como neoplasias de “baixo risco”, estiveram associados a metástases e a mortes dos pacientes num número considerável de casos. Este prognóstico pior do que o esperado reforça a necessidade de encontrar opções terapêuticas. De relevo e com este objectivo, identificámos pela primeira vez nestes tumores mutações frequentes e em diferentes domínios no gene CYLD, contribuindo para um melhor entendimento da sua patogénese genética. Esta informação pode ser usada para identificar os tumores, contribuindo para um diagnóstico precoce e selecção atempada dos pacientes para terapêutica apropriada. Para além disso, cria oportunidades de investigação ou para o desenvolvimento de ensaios clínicos dirigidos a alvos terapêuticos específicos.

O presente relatório de prática de ensino supervisionada incide sobre o tema A participação portuguesa na Primeira Guerra Mundial: um exemplo prático, tema inserido no subdomínio das Aprendizagens Essenciais de História do 9º ano de escolaridade Hegemonia e declínio da influência europeia, subdomínio este integrado no domínio A Europa e o Mundo no Limiar do Século XX. Esta proposta didática vem no decorrer da prática de ensino supervisionada na Escola Básica Dr. António Augusto Louro durante o ano lectivo de 2022/2023, tendo sido aplicada em contexto de sala de aula. Com o tema A participação portuguesa na Primeira Guerra Mundial: um exemplo prático, pretendemos evidenciar a participação portuguesa na Grande Guerra, mormente nas trincheiras europeias, abordando o quotidiano dos soldados portugueses. Para tal, iremos observar a acção do Corpo Expedicionário Português no teatro de guerra europeu, com especial atenção para a forma de como os soldados fizeram face às condições duríssimas que a Primeira Guerra Mundial impôs. Para além da acção bélica inerente ao conflito, os expedicionários portugueses viram-se posicionados num sector marcado por fracas condições de higiene e pela severidade climatérica. Não obstante, tentaram distrair-se de variadas formas: frequentavam estaminets para beber a misteriosa cerveja, jogavam cartas, conversavam entre si, cantavam e jogavam futebol, um jogo desconhecido para a maioria dos trinchas. Os mais instruídos liam jornais ou escreviam cartas à família ou à mulher francesa cujo namoro ainda não se encontrava consumado. Durante a leccionação dos conteúdos programáticos, procurámos aplicar algumas teorias de aprendizagens abordadas ao longo do Mestrado em Ensino de História. Neste sentido, adoptámos a teoria da Aprendizagem Significativa, preconizada por David Ausubel, nomeadamente através de um método expositivo com perguntas dirigidas aos discentes e do uso de mapas conceptuais. Noutras estratégias didáticas, como a análise de documentos a partir da consulta do manual, a resolução de exercícios, a realização de fichas de avaliação com consulta do manual e trabalhos de pesquisa, foram aplicados os princípios ligados ao Ensino por Descoberta, desenvolvido por Jerome Bruner.

O presente Relatório da Prática de Ensino Supervisionada foi realizado no âmbito do Mestrado em Ensino de Artes Visuais no 3.º ciclo do Ensino Básico e no Ensino Secundário e reúne as informações e investigações relativas à intervenção didática desenvolvida na Escola Secundária José Gomes Ferreira, em Lisboa, na disciplina de Oficina de Multimédia B, numa turma do 12.º ano do curso Científico-Humanístico de Artes Visuais. Com base na análise do contexto educativo, escolar e curricular, bem como das teorias da psicologia da educação, da prática artística e das suas pedagogias específicas, desenvolveu-se o presente projeto pedagógico que visa abrir horizontes e promover a exploração de diferentes meios válidos de criação no campo das artes visuais, de modo a impulsionar as aprendizagens. Esta unidade didática intitula-se “Mundo Surreal na P’Arte Digital” e representa a necessidade de investigar e compreender as potencialidades da exploração de meios digitais na criação artística e a sua importância no desenvolvimento das capacidades criativas, intelectuais e digitais dos alunos. A criação deste projeto teve em consideração diversos objetivos. Primeiramente, procurou-se que os alunos explorassem e dominassem as ferramentas de criação num software de edição de imagens bidimensionais, permitindo uma adaptação ao meio digital. Posteriormente, pretendeu-se que aprendessem a realizar várias fases de um projeto multimédia, usufruindo das potencialidades do meio digital na sua criação artística. Inicialmente, adotou-se a estratégia de definir um exercício de exploração das ferramentas, no qual os alunos trabalharam em pares. Isto permitiu que dominassem as ferramentas por meio da prática e da colaboração, além de estimular o surgimento de ideias para o projeto final por meio de debates entre os colegas e da partilha de conhecimentos prévios. Num segundo momento, foi-lhes lançado o desafio de, individualmente, realizar um projeto de criação de uma montagem surreal tirando partido das diferentes possibilidades do meio digital. Os resultados finais foram bastante positivos, tendo sido alcançadas aprendizagens significativas que foram de encontro com os objetivos traçados para esta unidade didática.

Background and Aims The beneficial effect of Hepatitis C virus (HCV) eradication by direct antiviral agents (DAAs) on liver fibrosis is well defined. Despite this, the impact of viral eradication in both hepatic and extra-hepatic metabolic features is underreached. This systematic review aimed to synthesize the evidence on the impact of HCV eradication by DAAs on liver steatosis, carotid atherosclerosis, glucidic impairment, dyslipidaemia, and weight gain. Methods A systematic search of the existing literature (up to December 2022) identified 97 original studies that fulfilled the inclusion criteria. Results Whereas total cholesterol and low-density lipoprotein (LDL) seem to increase after viral eradication, the cardiovascular damage expressed as carotid plaques and intima-media thickness seems to improve. Otherwise, the effect on liver steatosis, glucidic homeostasis, and weight seems to be strictly dependent on the presence of baseline metabolic disorders. Conclusion Despite high heterogeneity and relatively short follow-up of included studies, we can conclude that the presence of metabolic risk factors should be strictly evaluated due to their impact on liver steatosis, glucidic and lipid homeostasis, and on weight gain to better identify patients at risk of liver disease progression despite the virus eradication.

The Care of Late-Stage Parkinsonism (CLaSP) study is a longitudinal, multicentre, prospective cohort study to assess the needs and provision of care for people with late-stage Parkinson's disease and their caregivers in six European countries. As a cross-sectional study within the CLaSP study, 509 people with Parkinson's disease completed the "Schedule-for-Meaning-in-Life-Evaluation" (SMiLE) questionnaire. We compared the results to those of a representative sample of healthy participants (n = 856). People with late-stage Parkinson's disease reported family, partnership and spirituality as the greatest areas of importance. Overall, they had lower SMiLE indices compared to healthy participants. People with late-stage Parkinson's disease rated the importance of core meaning in life areas (namely family, social relations and health) as significantly lower than the representative cohort and they also rated satisfaction as significantly lower in most areas. In conclusion, people with late-stage Parkinson's disease do have areas where they can find meaning, such as family, partnership and spirituality. However, they indicate a lack of fulfilment of their individual MiL, reflected by low satisfaction rates in the majority of meaning in life categories. The need for spiritual support for people with Parkinson's disease indicates the important role of chaplains to help people with Parkinson's disease maintain meaning in life.

O presente relatório de prática de ensino supervisionada, tem por objetivo demonstrar o nosso percurso enquanto professores estagiários na Escola Secundária de Pinhal Novo, no âmbito do Mestrado em Ensino da História no 3º Ciclo do Ensino Básico e no Ensino Secundário. Neste relatório, tentamos explicar de que modo a Escola, enquanto espaço primordial de primeiro grande contacto comunitário, favorece o estabelecimento de vivências mais próximas de uma Cidadania ativa. Considerando isto, posteriormente, explicamos de que modo a disciplina de História, pelas suas características metodológicas e pelas temáticas que estuda, permite o desenvolvimento de uma consciência cívica, relevando-se assim a sua importância social. Contudo, acreditamos que para seja possível a História (e outras disciplinas), auxiliarem na digna formação dos indivíduos da nossa sociedade, é necessário haver reformas e mudanças estruturais. Para isso, os professores têm o direito de poder exercer a sua profissão num contexto educativo que possibilite a plena concretização deste intento, até para ser possível haver uma certa coerência. Assim sendo, a proposta que aqui se apresenta comprova de que forma, nas aulas de História, neste caso de 9º ano, os alunos através do estudo de um conjunto de temáticas e do aprofundamento de algumas competências, desenvolvem uma consciência cívica, com um enfoque na sua vertente política e social.

Despite the success of antiretroviral cocktails, a cure for HIV-1 remains elusive. This is mainly due to the existence of persistent cellular reservoirs infected with non-transcriptional, latent HIV-1. An effective treatment against HIV-1 would target both active and latent HIV-1-infected cells, and eliminate them without harming non-infected cells. In order to achieve this, we have developed a therapeutic plasmid with a Tat/Rev-dependent toxin expression. This vector contains a HIV-1 transcriptional activator to specifically activate latent infected cells. We confirmed the expression of all vector components necessary to achieve our goal. We then assessed the potential of this therapeutic plasmid to specifically eliminate infected cells by transduction of a T-lymphoma cell line that has a HIV-1 latent provirus integrated in its genome. We demonstrated that our therapeutic plasmid is not only capable of reactivating latent HIV-1 expression by itself, but it also can induce cell death efficiently and specifically in HIV-1 infected cells. Cell death was 60% higher in infected cells than in non-infected cells, indicating that our therapeutic vector was able to fulfill the objective proposed in this work. Therefore, the data presented here represents a promising approach for the development of a gene therapy that could become a viable and successful treatment against HIV-1.

O objeto desta tese são os limites que incidem sobre a divulgação nominal da remuneração dos servidores pela Administração Pública, motivada pela necessidade de fomentar a transparência administrativa, diante do direito à proteção de dados pessoais. Para tanto, serão apresentados os princípios da publicidade (no Brasil) e da administração aberta (em Portugal), conjugados com o direito de acesso à informação pública, o princípio da transparência administrativa e os mecanismos de accountability, como fatores efetivadores da participação popular do processo de controle dos atos Administração Pública, em especial, sobre a gestão dos gastos públicos, que tem sido um dos principais argumentos utilizados pelos defensores da divulgação nominal da remuneração dos servidores pela Administração Pública, sob a justificativa de se evitar a corrupção. Por outro lado, será fixada a importância do direito à reserva da vida privada, como fator de proteção do núcleo pessoal dos indivíduos e sua relação com o direito à proteção de dados pessoais. Neste contexto, será realizada a análise analítico-comparativa da legislação portuguesa e brasileira relevantes, respetivamente a Lei de Acesso aos Documentos Administrativos – LADA e o Regulamento Geral sobre a Proteção de Dados – RGPD e a Lei de Acesso à Informação – LAI e a Lei Geral de Proteção de Dados – LGPD. Além disso, serão debatidos os pontos relativos à tensão e suposto conflito entre os princípios e direitos em questão, sob o crivo dos critérios de proporcionalidade

To understand how galaxies evolve, it is crucial to track their morphological changes and identify the underlying mechanisms that drive them. Of particular interest to parameterize the disk, is the scale height of the vertical distribution of objects, above and below the Galactic plane, describes the thickness of the disk. Although the study of thickness of the disk can be made with different objects (e.g stars), open clusters are often used as disk probes due to the precision with which their distances and ages can be determined, making them key astrophysical objects for studying the morphological changes and evolution of our Galaxy. The scale height of the spatial distribution of open clusters in the Milky Way exhibits a well known increase with age. This increase is usually attributed, in a vague way, to disc heating mechanisms similar to those that act on individual stars or that the disk was thicker in the past. In this work, we address the evolution of the scale height of open clusters from a different angle, as an effect of the disruption of clusters due to disc phenomena such as encounters with giant molecular clouds. We present a dynamical model that follows the orbits of open clusters and includes their disruption due to interactions with the disc and mass loss due to stellar evolution and evaporation. The results show that the proposed model is a viable mechanism to explain the evolution of the scale height of open clusters. Additionally, the model is also able to predict the total number of open clusters that survive with age, indicating that the timescale of the disruption is being successfully reproduced. We identify additional features that can be added in the future to improve the model, namely mass dependent disruption by giant molecular clouds and initial cluster velocities.

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How are scientists coping with misinformation and disinformation? Focusing on the triangle scientists/mis-disinformation/behaviour, this study aims to systematically review the literature to answer three research questions: What are the main approaches described in the literature concerning scientists’ behaviour towards mis-disinformation? Which techniques or strategies are discussed to tackle information disorder? Is there a research gap in including scientists as subjects of research projects concerning information disorder tackling strategies? Following PRISMA 2020 statement, a checklist and flow diagram for reporting systematic reviews, a set of 14 documents was analysed. Findings revealed that the literature might be interpreted following Wilson and Maceviciute’s model as creation, acceptance and dissemination categories. Crossing over these categories, we advanced three standing points to analyse scientists’ positions towards mis-disinformation: inside, inside-out and outside-in. The stage ‘Creation/facilitation’ was the least present in our sample, but ‘Use/rejection/acceptance’ and ‘Dissemination’ were depicted in the literature retrieved. Most of the literature approaches were about inside-out perspectives, meaning that the topic is mainly studied concerning communication issues. Regarding the strategies against the information disorder, findings suggest that preventive and reactive strategies are simultaneously used. A strong appeal to a multidisciplinary effort against mis-disinformation is widely present, but there is a gap in including scientists as subjects of research projects.

Seed-based revegetation is fundamental in global restoration efforts. When seed storage is mandatory, knowledge of storage conditions that reduce ageing is essential for effective conservation of viable seeds. However, inappropriate storage is still frequent, particularly in local restoration projects involving native species. We investigated how different storage conditions (temperature and relative humidity) affected seed viability of the Mediterranean shrubs Arbutus unedo and Myrtus communis. We aimed to identify which conditions reduced seed deterioration and to provide practitioners with practical storage guidelines to overcome the natural fluctuations in seed production of these two species. Seeds equilibrated at 15%, 30% and 60% relative humidity (RH) and sealed in airtight containers were stored at 5 °C or ambient temperature over one year. Additional seed lots were kept under ambient conditions, according to the usual local practice (open storage). Germination tests were performed prior to and after storage. Significant seed deterioration (80–100%) occurred in seeds equilibrated at 60% RH and stored at ambient temperature, and in seeds kept in open storage. The remaining treatments maintained high seed viability. Short seed longevity under ambient conditions was demonstrated for both species. To store seeds over one year, practitioners may either (i) equilibrate seeds up to 60% RH and store them in a refrigerator (5 °C), or (ii) equilibrate seeds up to 30% RH (preferably 15% RH for A. unedo) and store them at ambient temperature. All seeds should be sealed into airtight containers immediately after equilibrium. The effectiveness of the 5 °C-60% RH treatment demonstrated that procedures less demanding than international standards for short-term storage can still ensure seed viability. While contributing to the understanding of species responses to storage conditions, our findings can be applied to both species by local practitioners across the Mediterranean. Moreover, our approach can be adapted and extended to other restoration projects and native species, supporting specific improvements to seed storage protocols, thereby enhancing the cost-effectiveness and the capacity of restoration efforts.

This study aimed to characterize the antimicrobial resistance (AMR) and virulence profiles of 67 Escherichia coli isolates obtained from faecal samples of 77 wild mammals from 19 different species, admitted in two rescue and rehabilitation centers in Costa Rica. It was possible to classify 48% (n = 32) of the isolates as multidrug-resistant, and while the highest resistance levels were found towards commonly prescribed antimicrobials, resistance to fluoroquinolones and third generation cephalosporins were also observed. Isolates obtained from samples of rehabilitated animals or animals treated with antibiotics were found to have significantly higher AMR levels, with the former also having a significant association with a multidrug-resistance profile. Additionally, the isolates displayed the capacity to produce α-haemolysins (n = 64, 96%), biofilms (n = 51, 76%) and protease (n = 21, 31%). Our results showed that AMR might be a widespread phenomenon within Costa Rican wildlife and that both free-ranging and rehabilitated wild mammals are potential carriers of bacteria with important resistance and virulence profiles. These results highlight the need to study potential sources of resistance determinants to wildlife, and to determine if wild animals can disseminate resistant bacteria in the environment, potentially posing a significant threat to public health and hindering the implementation of a “One Health” approach.