Molecular detection of Enterocytozoon bieneusi in bats from Portugal.

Enterocytozoon bieneusi is a microsporidia commonly found in the gastrointestinal tract of humans and a wide range of other animals, constituting a major cause of microsporidiosis in humans. Although E. bieneusi has been detected in humans, domestic, and wild animals in Portugal, and its presence in bats has been linked to zoonotic characteristics, its occurrence in bats within the country has not been reported. In this study, we investigated the presence of E. bieneusi in 380 bat fecal samples collected in mainland Portugal through a nested PCR assay targeting the internal transcribed spacer region and the flanking small and large subunits of the ribosomal RNA. Enterocytozoon bieneusi was detected in one bat sample (i.e., 0.26%; Pipistrellus pipistrellus). Additionally, another sample tested positive for Enterocytozoon sp. Phylogenetic analysis of the obtained ITS sequence of E. bieneusi revealed clustering within the potentially zoonotic Group 1. This study represents the first report of E. bieneusi in a bat from Europe. Findings presented here contribute to an enhanced understanding of E. bieneusi epidemiology.

Enterocytozoon bieneusi is the most frequent cause of microsporidiosis in humans. In this study, E. bieneusi, belonging to a potentially zoonotic Group, was detected in 0.26% bat samples from Portugal, highlighting bats' potential role in transmitting this microsporidia to humans and other animals.

First report of Enterocytozoon bieneusi in Erinaceus europaeus and Hemiechinus auritus hedgehogs from recovery centers of Portugal.

Enterocytozoon bieneusi microsporidia are emerging pathogens infecting a wide range of vertebrate and invertebrate hosts, known to have zoonotic features since they infect both wild and domestic animals, and humans. Despite their significance, there is very limited epidemiological data on microsporidia in hedgehogs, especially European hedgehogs (Erinaceus europaeus) and long-eared hedgehogs (Hemiechinus auritus), the former known as synantropic hedgehogs, and the latter suited as pets. As such, the present study aimed to assess the presence of E. bieneusi in hedgehogs from Portugal. For this purpose, fecal samples from 110 hedgehogs of three species-E. europaeus (n = 106), H. auritus (n = 1), and Atelerix albiventris (n = 3)-were collected and tested for E. bieneusi by PCR targeting the internal transcribed spacer region and the flanking small and large subunits of the rRNA. We found an overall occurrence of 22.7% (25/110; 95% confidence interval [CI]: 15.28-31.70), with 22.6% (24/106; 95% [CI]: 15.08-31.79) in E. europaeus, 100% (1/1) in H. auritus, and 0% in A. albiventris. Interestingly, three novel genotypes were identified, all belonging to the potentially zoonotic Group 1. Our findings highlight the importance of hedgehogs as potential reservoirs for E. bieneusi and emphasize the need for further research to understand their role in transmission dynamics and assess the associated risks to public and veterinary health.

Synanthropic hedgehogs were tested for Enterocytozoon bieneusi, the main cause of human microsporidiosis. Results showed 22.7% of hedgehogs were shedding E. bieneusi spores, with three new genotypes from the zoonotic Group 1. Hedgehogs may transmit to humans/animals, warranting more research.

Search for new biomarkers of tolerance to Perkinsus olseni parasite infection in Ruditapes decussatus clams.

The grooved carpet shell (Ruditapes decussatus) is a clam species with high economic and social importance in several European and Mediterranean countries. Production of this species suffered a decline caused by biotic (parasite infection) and abiotic factors (environmental factors, stress, poor management methods and intensive culture of the introduced species Ruditapes philippinarum). The protozoan parasite Perkinsus olseni is also responsible for the decline of production, being nowadays one of the major issues for clam culture. Molecular biomarkers that might represent tolerance of R. decussatus to P. olseni have already been uncovered, shedding light in a possible production improvement by selecting those clams with a strongest immune response. In the present study, new tolerance biomarkers to P. olseni infection in R. decussatus were identified. The haemolymph proteomic profiles of naturally non/low-infected (tolerant) and highly-infected (susceptible) clams by the parasite across several heavy affected areas of Europe were characterized through a shotgun proteomics approach. Also, the mechanisms that might be involved in the responses against the disease in chronic infections were explored. Proteins related to energy restoration and balance, metabolic regulation, energy accumulation, ROS production, lysosomal activity, amino acid synthesis, proteolytic activity, iron regulation, iron withholding, and immune response modulation were significantly regulated in susceptible clams. In the tolerant group, proteins related to phagocytosis regulation, control of cell growth and proliferation, gonadal maturation, regulation of apoptosis, growth modulation, response to oxidative stress, iron regulation, shell development and metabolic regulation were significantly expressed. In summary, the protein expression profile of tolerant individuals suggests that an efficient pathogen elimination mechanism coupled to a better metabolic regulation leads to a tolerance to the parasite infection by limiting the spread through the tissues.

Effect of the alveolate parasite Perkinsus olseni infection on sexual maturation and spawning efficiency of the clam Ruditapes decussatus.

The effect of Perkinsus olseni infection on the reproduction ability of clams has been underestimated so far. Although some studies found evidence of reduction of egg production and delay in gonad maturation after infection, the total effect of the infection is still unclear. In this study, Ruditapes decussatus clams from a naïve population were injected with two different doses of P. olseni parasites, a low dose leading to a light infection and a high dose leading to a heavy infection. Clams were maintained during 2 months for maturation, and at the end of the experiment, the spawning was induced, the number of larvae release and mortality were evaluated. During the maturation period, infection level, gonadal stage, condition index, gross biochemical composition and oxidative status of progenitors were evaluated at days 0, 30 and 60 post-injection. The effects of P. olseni infection on clams showed alterations on biochemical parameters, namely lipid peroxidation, a significant mortality and a delayed gonad maturation, with a greater effect in the highly infected individuals. The reproductive capacity of the clams was impaired in both infected groups showing a lower production and a higher mortality rate of larvae. Finally, this study indicates that the production of natural beds with a high prevalence of P. olseni could be compromised by a deregulation of the natural reproduction cycle and a decrease in larvae production by infected animals, probably due to a combination of lower egg production and lower lipid reserves in larvae from infected clams.

Evaluation of the Ruditapes decussatus immune response after differential injected doses of Perkinsus olseni.

Ruditapes decussatus is a highly appreciated clam species, with a high demand, especially in European countries. However, natural populations of this species have been affected by over-exploitation, abiotic factors and also by diseases mainly caused by bacteria, virus and protozoans. Within the latter group of pathogens, Perkinsus spp. has been attracting attention for causing relevant mortality rates on bivalves around the world. From all species of this genus, Perkinsus olseni is one of the most impactful for R. decussatus, as it may cause chronic infection with fatal consequences under adverse conditions. Therefore, in this study, naïve clams were injected with two doses of P. olseni in order to better understand the response of R. decussatus against this parasite. So, one of the main goals of this experiment was to evaluate the differential response of the clams at gene expression level after injection and also to monitor the possible mortalities related to parasite. Results indicated that the methods applied for injection and detection of parasite load are effective. Furthermore, altogether, R. decussatus gene expression studies indicate differences on expression of some of the addressed genes among distinct levels of infection over time, which require further studies to confirm if these genes could be used as molecular markers of P. olseni infection. According to the results obtained, Adiponectin-C1q seems to be a promising gene to be used as molecular marker of chronic infection in clams haemolymph.

The impact of the COVID-19 pandemic on young children and their caregivers.

BACKGROUND: The COVID-19 pandemic has adversely impacted child development and the well-being of caregivers, and such evidence ought to be used to inform public policy decisions. This study investigated the impact of COVID-19 on children's behaviours and their caregivers' needs. METHODS: A cross-sectional study was conducted with 153 caregivers of children (from 0 to 5 years old) from three public daycare centres in Brazil. The Nurturing Care Framework of the World Health Organization was used to guide the assessment of caregivers' needs. Online data collection using a questionnaire was conducted from June to July 2020. RESULTS: The COVID-19 pandemic increased stressors such as low family income, unemployment, sadness, depression and anxiety of caregivers. Their most commonly reported needs were related to offering age-appropriate playful activities (49.7%), organizing the care routine of children at home (41.8%) and educating children when they do something wrong (39.9%). Additionally, the results showed that misbehaviour, aggressiveness and agitation occurred more frequently among preschoolers than infants or toddlers (p ≤ 0.05). CONCLUSION: During the COVID-19 pandemic, public policies should provide mental health support to caregivers, as well as information about security, safety and early learning opportunities for childcare at home.

The complexity of having a child with a congenital heart defect in a developing country: A qualitative study of parental needs.

PURPOSE: To explore parental needs related to their experiences of living with a child with congenital heart defect (CHD) since the diagnosis. DESIGN AND METHODS: An interpretative qualitative study developed with nine parents of children between the ages of five months and 11 years diagnosed with CHD. Interviews were conducted at an ambulatory pediatric cardiology centre. Data were analyzed using inductive thematic analysis. The consolidated criteria for reporting qualitative research (COREQ) was followed for quality reporting. This research was approved by a research committee. RESULTS: One central theme emerged, namely 'A desire to feel safe in dealing with the demands of CHD,' along with two main themes. The first is 'Looking for effective relations with healthcare professionals and health care systems' which encompasses three types of need: (1) need for continuous, clear and accurate information; (2) need for resolution and the support of services such as the public health care system and social services; (3) need for trust in health care professionals. The second theme is 'Looking for balance in daily life' with two main needs: (1) maintain family functioning and (2) learn to deal with the child and CHD. CONCLUSION: The main parental needs are related to their interactions with healthcare professionals and healthcare systems, highlighted by a need for information and trust relationships to feel safe in their daily lives. PRACTICE IMPLICATIONS: Our results imply rethinking the nurse presence in ambulatory care, implementation of a family-centered care approach and addressing the diverse and multifaceted experiences and needs of parents and children with CHD in different health care contexts.

E-cadherin clone 36 nuclear staining dictates adverse disease outcome in lobular breast cancer patients.

Breast cancer is a heterogeneous disease and additional biomarkers for individually predicting patient outcomes are needed. Aberrant membrane E-cadherin immunoexpression has been demonstrated in lobular breast cancer. Also, E-cadherin nuclear staining has been reported, associating with prognosis in various tumors. Here, we explore whether membrane or nuclear staining of E-cadherin has the potential to dictate prognosis of patients with lobular breast cancer. We selected a cohort of 285 consecutively diagnosed lobular breast cancer patients and performed immunohistochemistry for E-cadherin (clones 36, EP700Y, and NCH38) and P-cadherin (clone 56C1) in representative formalin-fixed paraffin-embedded blocks. All patients were female, HER2-negative and surgically treated in a single institution. Survival curves were computed by Kaplan-Meier analysis. Hazard ratios and respective 95% confidence intervals were estimated using Cox regression models. Statistical significance was set at p < 0.05. Nuclear staining for E-cadherin clone 36 was frequent (35%), contrarily to other antibodies tested. Negative correlation was found between nuclear and membrane E-cadherin clone 36 immunostaining (rs = -0.30, p < 0.001), whereas positive correlation was found between membrane immunoexpression of E-cadherin clone 36 and P-cadherin (rs = 0.31, p < 0.001). Patients with any evidence of E-cadherin clone 36 nuclear immunostaining disclosed significantly worse overall survival, disease-specific-survival and disease/progression-free survival (hazard ratio = 2.059, 95% confidence interval 1.313-3.230; hazard ratio = 1.980, 95% confidence interval 1.121-3.495; and hazard ratio = 2.341, 95% confidence interval 1.403-3.905, respectively). Differences in survival were more remarkable when considering nuclear E-cadherin immunoexpression in ≥50% tumor cells. Poorer survival was maintained in multivariable analysis, after adjusting for age, menopausal and PR status, treatment course, vascular invasion, tumor grade and stage. Our results support the use of antibodies against the cytoplasmic domain of E-cadherin, such as clone 36, which may reveal nuclear immunostaining and indicate more aggressive clinical course in patients with lobular breast cancer. We hypothesize that E-cadherin is cleaved and translocated to nucleus functioning as transcription factor.

Pedobacter lusitanus sp. nov., isolated from sludge of a deactivated uranium mine.

Strain NL19T is a Gram-stain-negative, aerobic bacterium that was isolated from sludge of a deactivated uranium mine in Portugal. 16S rRNA gene sequence analysis revealed that strain NL19T is a member of the genus Pedobacter and closely related to the strains Pedobacter himalayensis MTCC 6384T, Pedobacter cryoconitis DSM 14825T, Pedobacter westerhofensis DSM 19036T and Pedobacterhartonius DSM 19033T. It had a DNA G+C content of 40.8 mol%, which agreed with the genus description. The main fatty acids included C16 : 1ω7c, C14 : 1ω5c, C4 : 0, iso-C17 : 0, iso-C17 : 0 3-OH, C16 : 0, anteiso-C15 : 0 and iso-C15 : 0 3-OH. The main lipids present were phospholipids (60 %) and sphingolipids (35 %). The most abundant phospholipids included phosphatidylethanolamine, phosphatidylinositol and phosphatidylcholine. Menaquinone-7 (MK-7) was the only isoprenoid quinone detected. DNA-DNA hybridization similarities between strain NL19T and Pedobacter himalayensis MTCC 6384T, Pedobacter cryoconitis DSM 14825T, Pedobacter westerhofensis DSM 19036T and Pedobacter hartonius DSM 19033T were 15.3 , 16.2 , 11.5 and 16.0 %, respectively. Strain NL19T can also be distinguished from these four species based on gyrB and intergenic transcribed spacers (ITS) sequences and by some phenotypic traits such as NaCl tolerance, pH, growth temperature and carbon source utilization. Strain NL19Trepresents a novel species of the genus Pedobacter, for which the name Pedobacter lusitanus sp. nov. is proposed. The type strain is NL19T (=LMG 29220T=CECT 9028T). An amended description of Pedobacter himalayensis is also included.

Self-efficacy scale for the establishment of good relationships with families in neonatal and pediatric hospital settings.

OBJECTIVE: The purpose of this study was to develop and test the psychometric properties of the Self-efficacy Scale for the Establishment of Good Relationships with Families in Neonatal and Pediatric Hospital Settings. METHOD: Methodological study grounded on self-efficacy theory was conducted in three phases: conceptual and operational definition (review of the literature and interviews with the target population), content validity (opinion of five experts e three clinical nurses), and exploratory factor analysis and internal consistency reliability (cross-sectional survey with a valid sample of 194 nurses). RESULTS: A ten-point Likert scale with 40-item was designed and one item was excluded after review by experts. Three factors emerged from the exploratory factor analysis. The Cronbach's alpha for all items was 0.983 with item-total correlations in the range 0.657 to 0.847. Cronbach's alpha value if item deleted were less than or equal to 0.983. CONCLUSION: The final version of the scale demonstrated psychometric adequacy. It is a useful tool to be administered in the clinical, educational and research nursing fields to measure nurses' self-efficacy beliefs concerning the establishment of good relationships with families. OBJETIVO: El propósito de este estudio fue desarrollar y probar las propiedades psicométricas de la Escala de Autoeficacia para el Establecimiento de Buenas Relaciones con las Familias en Ambientes Neonatales y Pediátricos. MÉTODO: Estudio metodológico fundamentado en la teoría de la auto-eficacia se realizó en tres fases: conceptual y definición operacional (revisión de la literatura y entrevistas con la población objetivo), la validez de contenido (opinión de cinco expertos y tres enfermeras clínicas), y el factor de análisis exploratorio e fiabilidad interna de consistencia (estudio transversal con una muestra válida de 194 enfermeras). RESULTADOS: La escala de Likert de diez puntos con 40 ítems fue diseñada y un elemento fue excluido después de la revisión por expertos. Hay tres factores que emergieron del análisis factorial exploratorio. El alfa de Cronbach para todas las partidas fue 0,983, con correlaciones ítem-total en el rango de 0,657 a 0.847. El Valor alfa de Cronbach excluyendo el ítem quedo inferior o igual a 0.983. CONCLUSIÓN: La versión final de la escala demostró psicometría adecuada. Se trata de una herramienta útil para ser administrada en los campos clínicos, educativos y de enfermería de investigación para medir las creencias de autoeficacia de enfermeros sobre el establecimiento de buenas relaciones con las familias.

Ecotoxicity and genotoxicity of a binary combination of triclosan and carbendazim to Daphnia magna.

In the environment, chemical substances appear as complex mixtures and consequently organisms are exposed to a variety of chemicals from different sources (e.g. wastewater treatment plants, agriculture runoffs). When studying chemical mixtures, there are two conceptual models usually used to predict toxicity: the Independent Action (IA) and Concentration Addition (CA) models. However, deviations from these reference models can occur as synergism or antagonism, dose ratio or dose level dependency. The aim of the present study was to investigate the effects of triclosan and carbendazim, and their binary mixture to Daphnia magna. With this purpose, immobilisation, feeding inhibition, and reproduction were assessed as main ecotoxicity endpoints. In addition, in vivo genotoxicity of both chemicals was investigated using the comet assay. In the single exposure, carbendazim was more toxic to D. magna than triclosan. When daphnids were exposed to both single compounds, DNA damage was observed. Concerning mixture exposures, different endpoints followed different patterns of response, from additivity: IA model (feeding inhibition and reproduction data), to deviations that indicate interaction between chemicals inside the organism: dose level dependency (immobilisation data) and dose ratio dependency (DNA damage). This study showed that additivity does not rule the dose-effect relation in chemical mixtures of carbendazim and triclosan and interactions between both chemicals might induce generally higher toxicity than predicted based on single chemical exposures.

A microcosm approach to evaluate the degradation of tributyltin (TBT) by Aeromonas molluscorum Av27 in estuarine sediments.

Tributyltin (TBT) is a biocide extremely toxic to a wide range of organisms, which has been used for decades in antifouling paints. Despite its global ban in 2008, TBT is still a problem of great concern due to the high levels trapped in sediments. Aeromonas molluscorum Av27 is a TBT degrading bacterium that was isolated from an estuarine system. We investigated the ability and the role of this bacterium on TBT degradation in this estuarine system, using a microcosm approach in order to mimic environmental conditions. The experiment was established and followed for 150 days. Simultaneously, changes in the indigenous bacterial community structure were also investigated. The results revealed a maximum TBT degradation rate of 28% accompanied by the detection of the degradation products over time. Additionally, it was observed that TBT degradation was significantly enhanced by the presence of Av27. In addition a significantly higher TBT degradation occurred when the concentration of Av27 was higher. TBT degradation affected the bacterial community composition as revealed by the changes in the prevalence of Proteobacteria subdivisions, namely the increase of Deltaproteobacteria and the onset of Epsilonproteobacteria. However, the addition of Av27 strain did not affect the dominant phylotypes. Total bacterial number, bacterial biomass productivity, 16S rRNA gene and denaturing gradient gel electrophoresis (DGGE) analyses also indicated alterations on the bacterial community structure over time, with bacteria non-tolerant to pollutants increasing their representativeness, as, for instance, the increase of the number of Alphaproteobacteria clones from 6% in the beginning to 12% at the end of the experiment. The work herein presented confirms the potential of Av27 strain to be used in the decontamination of TBT-polluted environments.

Nurses' attitudes regarding the importance of families in pediatric nursing care.

Affective, cognitive and behavioral components affect nurses´ attitudes to include families in the care processes. The purpose of this study was to investigate the attitudes of nurses about the importance of including families in nursing care. Data collection was performed in pediatric and maternal-child unit of a Brazilian university hospital. A sample of 50 nurses completed the Portuguese version of the instrument Families'Importance in Nursing Care-Nurses' Attitudes (FINC-NA). The results indicated that nurses have supportive attitudes regarding families participation in nursing care. Attitudes of lower support for involving families in nursing care were found among nurses with older age, more time in the profession and who had no previous contact with contents related to Family Nursing. The application of the instrument in other contexts of assistance may help to illuminate important aspects of the challenges to implementing a family-centered approach in clinical practice.

[Use of Gadolinium in Follow-Up MRI of Multiple Sclerosis Patients: Current Recommendations]. / Utilização de Gadolínio nas Ressonâncias Magnéticas de Controlo em Doentes com Esclerose Múltipla: Recomendações Atuais.

Multiple sclerosis is the most frequent demyelinating disease of the central nervous system and is characterized by early onset and progressive disability. Magnetic resonance imaging, due to its high sensitivity and specificity in the detection of demyelinating lesions, is the most useful diagnostic test for this disease, with the administration of gadolinium-based contrast agents being an important contribution to imaging interpretation. Although contrast is essential for diagnostic purposes, its routine use in monitoring disease activity, response to treatment, and related complications is controversial. This article aims to collate current recommendations regarding the use of gadolinium in the imaging follow-up of multiple sclerosis and establish effective and safe guidelines for clinical practice. The literature review was conducted in PubMed, using the terms 'multiple sclerosis', 'magnetic resonance imaging' and 'gadolinium', or 'contrast media'. Articles published between January 2013 and January 2023 concerning the safety of gadolinium and the use of these contrast agents in follow-up scans of adult patients diagnosed with multiple sclerosis were selected. Although no biological or clinical consequences have been unequivocally attributed to the retention of gadolinium in the brain, which were mostly reported with linear agents, health authorities have been recommending the restriction of contrast to essential clinical circumstances. In multiple sclerosis, the detection of subclinical contrast-enhancing lesions with no corresponding new/ enlarging T2-WI lesions is rare and has a questionable impact on therapeutic decisions. On the other hand, gadolinium has a higher sensitivity in the differential diagnosis of relapses, in the detection of recent disease activity, before and after treatment initiation, and in patients with a large lesion burden or diffuse/confluent T2-WI lesions. Contrary to progressive multifocal leukoencephalopathy screening, monitoring of immune restitution inflammatory syndrome also benefits from the administration of gadolinium. It is feasible and safe to exclude gadolinium-based contrast agents from routine follow-up scans of multiple sclerosis, despite their additional contribution in specific clinical circumstances that should be acknowledged by the neurologist and neuroradiologist.

A esclerose múltipla é a doença desmielinizante do sistema nervoso central mais frequente, caracterizando-se pelo início precoce e incapacidade progressiva. A ressonância magnética, pela elevada sensibilidade e especificidade na deteção de lesões desmielinizantes, é o exame complementar mais útil nesta patologia, sendo a administração de meios de contraste com gadolínio um importante contributo na interpretação imagiológica. Embora o contraste seja imprescindível no âmbito do diagnóstico, a sua utilização por rotina na monitorização da atividade de doença, resposta ao tratamento e respetivas complicações é controversa. O objetivo deste artigo é reunir as recomendações atuais relativas à utilização do gadolínio no seguimento imagiológico da esclerose múltipla e definir um protocolo clínico efetivo e seguro. A revisão da literatura foi conduzida na PubMed, recorrendo aos termos 'esclerose múltipla', 'ressonância magnética' e 'gadolínio' ou 'meio de contraste'. Foram selecionados artigos publicados entre janeiro de 2013 e de 2023 relativos à segurança do gadolínio e à sua utilização na ressonância magnética de controlo dos doentes adultos com diagnóstico de esclerose múltipla. Apesar de nenhuma consequência biológica ou clínica ter sido inequivocamente atribuída à retenção cerebral do gadolínio, que foi reportada maioritariamente com agentes lineares, as autoridades de saúde têm vindo a recomendar a restrição do contraste a circunstâncias clínicas essenciais. Na esclerose múltipla, a deteção de lesões subclínicas com captação de gadolínio sem tradução em lesões novas/aumentadas nas sequências ponderadas em T2 ocorre raramente e com impacto na decisão terapêutica questionável. Por outro lado, o gadolínio assume uma sensibilidade superior no diagnóstico diferencial de surtos clínicos, na deteção de atividade inflamatória recente, antes e após o início de uma terapêutica e nos doentes com elevada carga lesional ou lesões difusas/confluentes nas sequências ponderadas em T2. Contrariamente ao rastreio da leucoencefalopatia multifocal progressiva, a monitorização da síndrome inflamatória de reconstituição imunológica beneficia também da inclusão do gadolínio. É exequível e segura a exclusão do gadolínio no seguimento imagiológico de rotina da esclerose múltipla, apesar do seu contributo adicional em circunstâncias clínicas específicas que devem ser do conhecimento articulado do neurologista e neurorradiologista.

Detection of Enterocytozoon bieneusi in Non-Human Primates in Portuguese Zoos.

Enterocytozoon bieneusi, an intracellular eukaryote closely related to fungi, is recognized as a significant pathogen affecting humans, particularly those with compromised immune systems. While its transmission routes are still not fully elucidated, fecal-oral transmission remains the primary one. With a wide host range, the zoonotic potential of E. bieneusi is a concern, albeit direct evidence of animal-to-human transmission remains scarce. Genotyping based on the internal transcribed spacer (ITS) region facilitates the delineation of genetic diversity, with potentially zoonotic genotypes predominantly associated with Groups 1 and 2. Despite the broad spectrum of susceptible animal hosts, research into microsporidian infection among zoo animals remains limited. This study aimed to evaluate the occurrence of E. bieneusi infection across diverse captive animals, focusing on zoo settings in Portugal. Fecal samples were collected from a variety of animals, and molecular detection of E. bieneusi was conducted using nested PCR targeting the ITS region. Of 127 fecal samples, 1.57% (95% CI: 0.19-5.57) tested positive for E. bieneusi, with non-human primates (NHP's) exhibiting an 18.18% (95% CI: 2.28-51.78) occurrence. Phylogenetic analysis revealed clustering within Group 2 genotypes, indicating potential zoonotic implications. This study highlights the need for further research to understand the epidemiology of E. bieneusi in zoo environments and its potential transmission pathways to humans.

A Thorny Tale of Parasites: Screening for Enteric Protozoan Parasites in Hedgehogs from Portugal.

Enteric protozoan parasites, such as Blastocystis sp., Balantioides coli, Cryptosporidium spp., and Giardia duodenalis, may have implications for both animal and human health.Transmitted through the fecal-oral route, these parasites cause symptoms such as diarrhea, abdominal pain, and weight loss. This study investigated the presence of these enteric protozoan parasites and genetically characterized them in hedgehogs from Portugal. A total of 110 hedgehog stool samples were collected. Molecular detection methods showed an overall occurrence of protozoa in 1.82% (2/110 95% CI: 0.22-6.41) of hedgehogs, with Blastocystis being found in one hedgehog and Cryptosporidium being found in another. No evidence for the presence of B. coli or G. duodenalis was found. This study suggests that there is a need to stay aware of hedgehogs as potential hosts of enteric protozoa. Ongoing research and surveillance efforts are recommended to explore practical prevention and control strategies. The results contribute to the limited knowledge of these parasites in Portuguese hedgehog populations and underscore their potential relevance to both veterinary and public health.

Serological and Molecular Survey of Hepatitis E Virus in Small Ruminants from Central Portugal.

Hepatitis E virus (HEV) is currently recognized as an emerging problem and a growing concern for public health in developed countries, with HEV infections mainly attributable to foodborne transmission of HEV-3. The zoonotic HEV genotype 3 infects a wide range of mammalian hosts, with swine considered as the primary host. This study investigates the occurrence of HEV among small ruminants in Portugal. The primary aim of the present research was to evaluate the circulation and the potential for HEV infection among sheep and goats. A total of 400 bile samples and 493 blood samples were collected from sheep and goats at a slaughterhouse in the center region of Portugal, between January 2022 and March 2023. The HEV RNA detection in bile samples was performed using a nested broad-spectrum RT-PCR targeting the ORF1 region. Serological analysis to detect anti-HEV antibodies was conducted using a commercial double-antigen sandwich multi-species ELISA. The HEV RNA was not detected in any bile samples using the nested broad-spectrum RT-PCR. Serological analysis revealed an overall HEV antibody seroprevalence of 2% (10/493, 95% CI: 0.98-3.70) among the small ruminants, namely 2.2% in goats and 2.0% in sheep. Curiously, no statistically significant association among the factors, age, sex and species and HEV seroprevalence was observed. Although HEV RNA was not detected in the bile of sheep and goats, this study the evidence of seroprevalence in these small ruminant species. Further research could provide additional insights into the factors influencing HEV transmission dynamics in small ruminants in Portugal and its potential implications for public health.

Molecular evidence of sporadic Coxiella burnetii excretion in sheep milk, central Portugal.

Coxiella burnetii is the etiologic agent of Q fever, a worldwide zoonosis. Cattle, sheep and goats are considered the main reservoirs of the disease. Transmission to humans occurs mainly through the inhalation of infectious aerosols from milk, faeces, urine, and birth products from infected ruminants. In this study, a 2-year longitudinal approach was performed to ascertain the excretion of C. burnetii in bulk tank milk samples of sheep from a mountain plateau in central Portugal, with sampling conducted during the years 2015 and 2016. From a total of 156 bulk tank milk samples tested by qPCR, only one showed to be positive for C. burnetii (1.28% [95%CI: 0.03-6.94]), from 2015, the first year of collection. Bidirectional sequencing and phylogenetic analysis of IS1111 transposase partial region confirmed the presence of C. burnetii DNA. The presence of C. burnetii in raw milk samples highlights the necessity for additional research to determine if raw milk is a potential source for human infection. Animal health surveillance and prevention measures against this zoonotic disease should be considered.

Aeromonas molluscorum Av27 is a potential tributyltin (TBT) bioremediator: phenotypic and genotypic characterization indicates its safe application.

Aeromonas molluscorum Av27 is an estuarine bacterium highly resistant to tributyltin (TBT). Also, the strain is able to degrade TBT into the less toxic compounds dibutyltin and monobutyltin. Therefore, this bacterium has potential to be employed in bioremediation processes. In this context, defining its biological safety is crucial. With that purpose a number of intrinsic characteristics, usually present/associated with virulent strains, were investigated. Few virulence factors were detected in strain Av27. For instance, a DNase gene is present, but it is not apparently expressed in vitro. Motility, adherence factor and phospholipase activity were also detected. Additionally, cytotoxicity to Vero cells was negative. Resistance to penicillin (10 µg ml(-1)), amoxicillin/clavulanic acid (30 µg ml(-1)) and cephalothin (30 µg ml(-1)) and also to the vibriostatic agent O/129 was observed. Five plasmids (4, 7, 10, 100 kb and one greater than 100 kb) were identified. No Class I and II integrons were detected. Study of the optimal growth conditions showed that Av27 easily adapts to different environmental conditions. Overall, the results suggest that A. molluscorum Av27 can be considered safe to use to bioremediate TBT in contaminated environments.

Medication reconciliation in pediatrics: a validation of instruments to prevent medication errors.

OBJECTIVES: to develop and validate the content of two instruments for promoting medication reconciliation for the transition of care of hospitalized children. METHODS: methodological study, conducted in five stages: scope review for conceptual structure; elaboration of the initial version; content validation with five specialists using the Delphi technique; reassessment; and construction of the final version of the instruments. A content validity index of at least 0.80 was adopted. RESULTS: three rounds of evaluation were carried out to reach the validity index of the proposed contents, whereas a new analysis of 50% of the 20 items of the instrument aimed at families, and 28.5% of the 21 items aimed at professionals was necessary. The instrument aimed at families reached an index of 0.93, and the instrument for professionals, 0.90. CONCLUSIONS: the proposed instruments were validated. It is now possible to proceed with practical implementation studies to identify their influence on safety during medication reconciliation at transition of care.