Occurrence of microplastics and metals in European seabass produced in different aquaculture systems: Implications for human exposure, risk, and food safety.

Microplastics (MPs) are emerging contaminants of increasing concern as they may cause adverse effects and carry other contaminants, which may potentially compromise human health. Despite occurring in aquatic ecosystems worldwide, the knowledge about MP presence in different aquaculture systems and their potential impact on seafood products is still limited. This study aimed to determine the levels of MPs in water, feed, and European seabass (Dicentrarchus labrax) from three relevant aquaculture systems and estimate human exposure to MPs and metals through seabass consumption. The recirculating aquaculture system (RAS) had the highest MP occurrence in water and feed. MP levels in seabass followed the aquaculture system's levels in water and feed, with RAS-farmed fish presenting the highest MP load, both in the fish gastrointestinal tract (GIT) and muscle, followed by pond-, and cage-farmed fish. MPs' characteristics across aquaculture systems and fish samples remained consistent, with the predominant recovered particles falling within the MP size range. The particles were visually characterized and chemically identified by micro-Fourier Transform Infrared Spectroscopy (µFTIR). Most of these particles were fibres composed of man-made cellulose and PET. MP levels in GIT were significantly higher than in muscle for pond- and RAS-farmed fish, MPs' bioconcentration factors >1 indicated bioconcentration in farmed seabass. Metal concentrations in fish muscle were below permissible limits, posing low intake risks for consumers according to the available health-based guidance values and estimated dietary scenarios.

Enhancing the authenticity of animal by-products: harmonization of DNA extraction methods from novel ingredients.

Introduction: The increasing global pressure to explore alternative protein sources derived from animal by-products has opened-up opportunities, but it has also created the need to assess their compliance with labelling statements, to ensure consumer's trust in the composition of both feed and food products. Assessing the authenticity of highly processed animal by-products, particularly within the rapidly expanding Halal food market, presents a significant challenge due to the lack of robust and standardized methodologies. However, the success of DNA based authenticity system is highly dependent on the extracted DNA quantity, quality, and purity ratios from heterogeneous matrices. Material and methods: In this work, nine DNA extraction methods were tested on selected processed animal by-products with high-value and interest for the feed industry: meals from poultry meat, blood and feather, and hydrolysates from swine meat and bone, fish, and black soldier fly. The proposed DNA extraction methods are developed to specifically target swine-specific mitochondrial region, as a case study. Results and discussion: Both the conventional CTAB method and the commercial kits, specifically Invisorb® Spin Tissue Mini and NucleoSpin™ Food, demonstrated superior extraction efficiency and quality ratios. Nevertheless, commercial kits enabled faster detection in comparison to the conventional methods. The absence of swine DNA was successfully validated and confirmed in all animal meals and hydrolysates that did not contain swine in their composition beforehand, demonstrating their compliance with the Halal market requirements.

Escherichia coli Phylogenetic and Antimicrobial Pattern as an Indicator of Anthropogenic Impact on Threatened Freshwater Mussels.

Freshwater bivalves are widely used as accumulation indicators and monitoring tools for assessing contaminant effects on different levels of biological integration. This pilot study aimed to explore the phylogenetic diversity of Escherichia coli isolated from freshwater mussels (Margaritifera margaritifera and Potomida littoralis) and characterize their phenotypes and antibiotic resistance profiles. Samples were collected in the Rabaçal and Tua Rivers, in the Douro basin, Portugal-two sites representing different levels of anthropogenic contamination. Antimicrobial susceptibility testing was performed via the disk diffusion method with 21 antibiotics. Results showed that 31% of isolates were multidrug-resistant (MDR). Thus, freshwater mussels provide an effective and time-integrated approach for identifying/quantifying fecal indicators, including MDR bacteria. PCR-based assays were designed for assessing phylogenetic E. coli groups. Among the E. coli isolates, the highest prevalence (44%) was observed in group D or E, followed by group E or Clade I (25%), group A (19%), and group B1 (13%). E. coli isolated from M. margaritifera predominantly exhibited a higher prevalence of phylogroups D or E, whereas E. coli from P. littoralis showed associations with phylogroups E or clade I, B1, A, and D or E. Our results provide new insights into the phylogenetic diversity of E. coli in freshwater bivalves. Additionally, the findings highlight the possible linkage of phylogroups with the host species, the geographical location in the water stream, and human activity. Using E. coli as a bioindicator isolated from freshwater mussels helps us grasp how human activities affect the environment. This study has important implications for those interested in safeguarding water resources, especially in tackling antibiotic resistance in aquatic ecosystems.

Genetic Ablation of Inositol 1,4,5-Trisphosphate Receptor Type 2 (IP3R2) Fails to Modify Disease Progression in a Mouse Model of Spinocerebellar Ataxia Type 3.

Spinocerebellar ataxia type 3 (SCA3) is a rare neurodegenerative disease caused by an abnormal polyglutamine expansion within the ataxin-3 protein (ATXN3). This leads to neurodegeneration of specific brain and spinal cord regions, resulting in a progressive loss of motor function. Despite neuronal death, non-neuronal cells, including astrocytes, are also involved in SCA3 pathogenesis. Astrogliosis is a common pathological feature in SCA3 patients and animal models of the disease. However, the contribution of astrocytes to SCA3 is not clearly defined. Inositol 1,4,5-trisphosphate receptor type 2 (IP3R2) is the predominant IP3R in mediating astrocyte somatic calcium signals, and genetically ablation of IP3R2 has been widely used to study astrocyte function. Here, we aimed to investigate the relevance of IP3R2 in the onset and progression of SCA3. For this, we tested whether IP3R2 depletion and the consecutive suppression of global astrocytic calcium signalling would lead to marked changes in the behavioral phenotype of a SCA3 mouse model, the CMVMJD135 transgenic line. This was achieved by crossing IP3R2 null mice with the CMVMJD135 mouse model and performing a longitudinal behavioral characterization of these mice using well-established motor-related function tests. Our results demonstrate that IP3R2 deletion in astrocytes does not modify SCA3 progression.

Microplastics in water, feed and tissues of European seabass reared in a recirculation aquaculture system (RAS).

Plastic particles (PLs) are ubiquitous in aquatic ecosystems, and aquaculture production is susceptible to contamination from external or endogenous sources. This study investigated PL presence in water, fish feed and body sites of 55 European seabass produced in a recirculating aquaculture system (RAS). Fish morphometric parameters and health status biomarkers were determined. A total of 372 PLs were recovered from water (37.2 PL/L), 118 PLs from feed (3.9 PL/g), and 422 from seabass (0.7 PL/g fish; all body sites analysed). All 55 specimens had PLs in at least two of the four body sites analysed. Concentrations were higher in the gastrointestinal tract (GIT; 1.0 PL/g) and gills (0.8 PL/g) than in the liver (0.8 PL/g) and muscle (0.4 PL/g). PL concentration in GIT was significantly higher than in muscle. Black, blue, and transparent fibres made of man-made cellulose/rayon and polyethylene terephthalate were the most common PLs in water and seabass, while black fragments of phenoxy resin were the most common in feed. The levels of polymers linked to RAS components (polyethylene, polypropylene, and polyvinyl chloride) were low, suggesting a limited contribution to the overall PL levels found in water and/or fish. The mean PL size recovered from GIT (930 µm) and gills (1047 µm) was significantly larger than those found in the liver (647 µm) and dorsal muscle (425 µm). Considering all body sites, PLs bioconcentrated in seabass (BCFFish >1), but their bioaccumulation did not occur (BAFFish <1). No significant differences were observed in oxidative stress biomarkers between fish with low (<7) and high (≥7) PL numbers. These findings suggest that fish produced in RAS are mainly exposed to MPs through water and feed. Further monitoring under commercial conditions and risk assessment are warranted to identify potential threats to fish and human health and define mitigating measures.

Astrocyte structural heterogeneity in the mouse hippocampus.

Astrocytes are integral components of brain circuits, where they sense, process, and respond to surrounding activity, maintaining homeostasis and regulating synaptic transmission, the sum of which results in behavior modulation. These interactions are possible due to their complex morphology, composed of a tree-like structure of processes to cover defined territories ramifying in a mesh-like system of fine leaflets unresolved by conventional optic microscopy. While recent reports devoted more attention to leaflets and their dynamic interactions with synapses, our knowledge about the tree-like "backbone" structure in physiological conditions is incomplete. Recent transcriptomic studies described astrocyte molecular diversity, suggesting structural heterogeneity in regions such as the hippocampus, which is crucial for cognitive and emotional behaviors. In this study, we carried out the structural analysis of astrocytes across the hippocampal subfields of Cornu Ammonis area 1 (CA1) and dentate gyrus in the dorsoventral axis. We found that astrocytes display heterogeneity across the hippocampal subfields, which is conserved along the dorsoventral axis. We further found that astrocytes appear to contribute in an exocytosis-dependent manner to a signaling loop that maintains the backbone structure. These findings reveal astrocyte heterogeneity in the hippocampus, which appears to follow layer-specific cues and depend on the neuro-glial environment.

Conocimientos, actitudes y prácticas enfermeras en materia de higiene bucal de los pacientes hospitalizados dependientes / Knowledge, attitudes and practices of nurses regarding oral hygiene of dependent inpatients

Objetivo: Valorar los conocimientos, actitudes y prácticas enfermeros en materia de higiene bucal en pacientes hospitalizados. Métodos: Estudio cuantitativo, descriptivo y transversal. Los datos se recogieron mediante un cuestionario autoadministrado, aplicado a 100 enfermeras de los servicios médicos de internación de dos hospitales del norte de Portugal, que evaluó tres dimensiones: conocimientos, actitudes y prácticas en materia de higiene bucal, que se resumieron en descripciones estadísticas que incluyen porcentajes, frecuencias, medias y desviaciones estándar, utilizando para el análisis de datos SPSS versión 23. Resultados: La puntuación media de conocimiento fue de 13,98 sobre 22, y la puntuación de las actitudes de los participantes hacia la higiene bucal fue de 48,35 sobre 60 puntos. Todos los participantes reconocen la importancia del cuidado de la higiene bucal para los pacientes hospitalizados, y el 96% asocian la falta de cuidado con la enfermedad sistémica. En cuanto a las prácticas, el 90% de los participantes evalúan la necesidad de cuidados de higiene bucal para las personas hospitalizadas en las primeras 24horas y el 61% documentan el resultado de esta evaluación. Conclusión: Los resultados muestran que, aunque los participantes son conscientes de la importancia de la salud bucal, los conocimientos y las prácticas no son consistentes. En cuanto a las prácticas de salud bucal, es urgente reducir la brecha entre la evidencia y la práctica y promover la estandarización del cuidado bucal.(AU)

Objective:Assess nurses’ knowledge, attitudes and practices towards oral hygiene of dependent inpatients. Methods: Quantitative, descriptive, and cross-sectional study. Data were collected through a self-administered questionnaire applied to 100 nurses from internal medicine wards of two hospitals in Northern Portugal, which assessed three dimensions: knowledge, attitudes, and practices regarding oral hygiene. Knowledge, attitudes, and practices in oral care were summarized in statistical descriptions including percentages, frequencies, means, and standard deviations using SPSS version 23 for data analysis. Results: The mean total knowledge score was 13.98 out of 22 and the participants’ mean score of the attitudes towards oral care was 48.35 out of 60 points. All participants acknowledge the importance of oral care for inpatients, with 96% associating poor oral hygiene with systemic disease. As for practices, 90% of participants assess the need for oral care of inpatients in the first 24hours, and 61% document the result of this assessment. Conclusion: The results show that although participants are aware of the importance of oral care, knowledge and practices are not consistent. Concerning oral health practices, it is urgent to narrow the gap between evidence and practice and promote oral care standardization.(AU)

Knowledge, attitudes and practices of nurses regarding oral hygiene of dependent inpatients.

OBJECTIVE: Assess nurses' knowledge, attitudes and practices towards oral hygiene of dependent inpatients. METHODS: Quantitative, descriptive, and cross-sectional study. Data were collected through a self-administered questionnaire applied to 100 nurses from internal medicine wards of two hospitals in Northern Portugal, which assessed three dimensions: knowledge, attitudes, and practices regarding oral hygiene. Knowledge, attitudes, and practices in oral care were summarized in statistical descriptions including percentages, frequencies, means, and standard deviations using SPSS version 23 for data analysis. RESULTS: The mean total knowledge score was 13.98 out of 22 and the participants' mean score of the attitudes towards oral care was 48.35 out of 60 points. All participants acknowledge the importance of oral care for inpatients, with 96% associating poor oral hygiene with systemic disease. As for practices, 90% of participants assess the need for oral care of inpatients in the first 24 h, and 61% document the result of this assessment. CONCLUSION: The results show that although participants are aware of the importance of oral care, knowledge and practices are not consistent. Concerning oral health practices, it is urgent to narrow the gap between evidence and practice and promote oral care standardization.

Calidad de vida y riesgos psicosociales de las enfermeras implicadas en la lucha contra la pandemia de Covid-19 / Quality of life and psychosocial risks of nurses involved in the battle against the COVID-19 pandemic

Objetivo: El objetivo de la presente investigación es comprender el impacto del cuidado de la persona sospechosa o infectada por el SARS-CoV-2, en la calidad de vida y en los factores de riesgo psicosociales de los enfermeros e identificar asociaciones con sus características sociodemográficas. Metodología: Se realizó un estudio exploratorio-descriptivo a través de las escalas World Health Organization Quality of Life Instruments - Bref y Copenhagen Psychosocial Questionnaire, con un total de 127 respuestas válidas. Los datos fueron trabajados en el SPSS, con recurso a la estadística descriptiva y al Crosstabs. Resultados principales: Los valores medios de la calidad de vida no alcanzaron los 70 puntos y se identificaron 22 factores de riesgo psicosociales en grado intermedio y elevado. Algunas características sociodemográficas parecen presentar asociación con estes resultados. Conclusión principal: Los enfermeros de la muestra se vieron afectados de manera significativa por la atención en el contexto de la pandemia.(AU)

Objective: The present investigation aims to understand the impact of providing care to people with suspected or confirmed SARS-CoV-2 infection on nurses' quality of life and psychosocial risk factors and to identify associations with their sociodemographic characteristics. Methods: An exploratory-descriptive study was carried out through the World Health Organization Quality of Life Instruments - Bref and Copenhagen Psychosocial Questionnaire scales, with a total of 127 valid responses. Data were processed in SPSS, using descriptive statistics and crosstabs. Results: The mean values of quality of life did not reach 70 points and 22 psychosocial risk factors were identified in intermediate and high degree. Some sociodemographic characteristics seem to be associated with these results. Conclusion: The nurses of the sample were significantly affected by the provision of care in the context of the pandemic.(AU)

Tet3 Deletion in Adult Brain Neurons of Female Mice Results in Anxiety-like Behavior and Cognitive Impairments.

TET enzymes (TET1-3) are dioxygenases that oxidize 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC) and are involved in the DNA demethylation process. In line with the observed 5hmC abundance in the brain, Tet genes are highly transcribed, with Tet3 being the predominant member. We have previously shown that Tet3 conditional deletion in the brain of male mice was associated with anxiety-like behavior and impairment in hippocampal-dependent spatial orientation. In the current study, we addressed the role of Tet3 in female mice and its impact on behavior, using in vivo conditional and inducible deletion from post-mitotic neurons. Our results indicate that conditional and inducible deletion of Tet3 in female mice increases anxiety-like behavior and impairs both spatial orientation and short-term memory. At the molecular level, we identified upregulation of immediate-early genes, particularly Npas4, in both the dorsal and ventral hippocampus and in the prefrontal cortex. This study shows that deletion of Tet3 in female mice differentially affects behavioral dimensions as opposed to Tet3 deletion in males, highlighting the importance of studying both sexes in behavioral studies. Moreover, it contributes to expand the knowledge on the role of epigenetic regulators in brain function and behavioral outcome.

Anogenital condylomata acuminata in young children: not always result of sexual transmission.

Condylomata acuminata lesions, commonly known as anogenital warts, are caused by human papillomavirus and manifest as flesh-coloured or hyperpigmented papules or plaques in the anogenital region. We report the case of a previously healthy young child that presented with anal condyloma lesions without other accompanying symptoms. His mother had similar lesions in the genital area. After careful investigation, no evidence of sexual abuse was found. Gradual clinical improvement was observed without treatment. This case highlights a disease that, when diagnosed in children, should alert the physician to the possibility of sexual abuse. However, other transmission routes, such as vertical transmission or autoinoculation and heteroinoculation during non-sexual contact, should also be considered.

Astrocyte-derived TNF and glutamate critically modulate microglia activation by methamphetamine.

Methamphetamine (Meth) is a powerful illicit psychostimulant, widely used for recreational purposes. Besides disrupting the monoaminergic system and promoting oxidative brain damage, Meth also causes neuroinflammation, contributing to synaptic dysfunction and behavioral deficits. Aberrant activation of microglia, the largest myeloid cell population in the brain, is a common feature in neurological disorders triggered by neuroinflammation. In this study, we investigated the mechanisms underlying the aberrant activation of microglia elicited by Meth in the adult mouse brain. We found that binge Meth exposure caused microgliosis and disrupted risk assessment behavior (a feature that usually occurs in individuals who abuse Meth), both of which required astrocyte-to-microglia crosstalk. Mechanistically, Meth triggered a detrimental increase of glutamate exocytosis from astrocytes (in a process dependent on TNF production and calcium mobilization), promoting microglial expansion and reactivity. Ablating TNF production, or suppressing astrocytic calcium mobilization, prevented Meth-elicited microglia reactivity and re-established risk assessment behavior as tested by elevated plus maze (EPM). Overall, our data indicate that glial crosstalk is critical to relay alterations caused by acute Meth exposure.

A Multidisciplinary Fingerprinting Approach for Authenticity and Geographical Traceability of Portuguese Wines.

The interest in developing reliable wine authenticity schemes is a hot-topic, especially for wines with recognized added-value. In order to accomplish this goal, two dimensions need to be considered: the grapevine variety determination and the geographical provenance. The aim of this study was to develop a multidisciplinary approach applicable to wines from the sub region Melgaço and Monção of the demarcated Vinho Verde region and from the demarcated Douro region. The proposed scheme consists on the use of DNA-based assays to detect Single Nucleotide Polymorphisms (SNPs) on three genes of the anthocyanin pathway (UFGT, F3H and LDOX) coupled with High-resolution melting (HRM) analysis aiming the varietal identification. The Alvarinho wines revealed to have the same haplotype using this marker set, demonstrating its applicability for genetic identification. In addition, to assess their geographical provenance, a multi-elemental approach using Sr and Pb isotopic ratios of wine, soil and bedrock samples was used. The isotopic data suggest a relation between Sr and Pb uptake by vine roots and soil's texture and clay content, rather than with the whole rock's isotopic ratios, but also highlights the potential of a discriminating method based on the combination of selected isotopic signatures.

Editorial Comments to the Special Issue: "Colletotrichum spp. on Fruit Crops-State of the Art, Perspectives and Drawbacks".

The year 2020 has been celebrated as the International Year of Plant Health by the United Nations, and it has been a unique opportunity to realise the vital role of producing while preserving our natural and cultural heritage-Sustainable Food and Agriculture [...].

Tet3 ablation in adult brain neurons increases anxiety-like behavior and regulates cognitive function in mice.

TET3 is a member of the ten-eleven translocation (TET) family of enzymes which oxidize 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC). Tet3 is highly expressed in the brain, where 5hmC levels are most abundant. In adult mice, we observed that TET3 is present in mature neurons and oligodendrocytes but is absent in astrocytes. To investigate the function of TET3 in adult postmitotic neurons, we crossed Tet3 floxed mice with a neuronal Cre-expressing mouse line, Camk2a-CreERT2, obtaining a Tet3 conditional KO (cKO) mouse line. Ablation of Tet3 in adult mature neurons resulted in increased anxiety-like behavior with concomitant hypercorticalism, and impaired hippocampal-dependent spatial orientation. Transcriptome and gene-specific expression analysis of the hippocampus showed dysregulation of genes involved in glucocorticoid signaling pathway (HPA axis) in the ventral hippocampus, whereas upregulation of immediate early genes was observed in both dorsal and ventral hippocampal areas. In addition, Tet3 cKO mice exhibit increased dendritic spine maturation in the ventral CA1 hippocampal subregion. Based on these observations, we suggest that TET3 is involved in molecular alterations that govern hippocampal-dependent functions. These results reveal a critical role for epigenetic modifications in modulating brain functions, opening new insights into the molecular basis of neurological disorders.

IP3 R2 null mice display a normal acquisition of somatic and neurological development milestones.

Astrocytes are key players in the regulation of brain development and function. They sense and respond to the surrounding activity by elevating their intracellular calcium (Ca2+ ) levels. These astrocytic Ca2+ elevations emerge from different sources and display complex spatio-temporal properties. Ca2+ elevations are spatially distributed in global (soma and main processes) and/or focal regions (microdomains). The inositol 1,4,5-trisphosphate receptor type 2 knockout (IP3 R2 KO) mouse model lacks global Ca2+ elevations in astrocytes, and it has been used by different laboratories. However, the constitutive deletion of IP3 R2 during development may trigger compensating phenotypes, which could bias the results of experiments using developing or adult mice. To address this issue, we performed a detailed neurodevelopmental evaluation of male and female IP3 R2 KO mice, during the first 21 days of life, as well as an evaluation of motor function, strength and neurological reflexes in adult mice. Our results show that male and female IP3 R2 KO mice display a normal acquisition of developmental milestones, as compared with wild-type (WT) mice. We also show that IP3 R2 KO mice display normal motor coordination, strength and neurological reflexes in adulthood. To exclude a potential compensatory overexpression of other IP3 Rs, we quantified the relative mRNA levels of all 3 subtypes, in brain tissue. We found that, along with the complete deletion of Itpr2, there is no compensatory expression of Itpr1 or Itrp3. Overall, our results show that the IP3 R2 KO mouse is a reliable model to study the functional impact of global IP3 R2-dependent astrocytic Ca2+ elevations.

Real-time PCR assay for Colletotrichum acutatum sensu stricto quantification in olive fruit samples.

Olive anthracnose is caused by fungal species within the Colletotrichum acutatum, C. gloeosporioides and C. boninense complexes. Anthracnose causes severe pre- and post-harvest olive drupe fall. This study aimed to design a species-specific qPCR assay, based on klap1 gene, suitable for C. acutatum s.s. quantification in cv. Galega Vulgar fruit samples. The developed qPCR assay presented a detection limit of 10.14 fg/reaction, and a linear cycle threshold of R2 = 0.996. C. acutatum inoculum was detected in pulverized olive fruits, and in early infection stages, before symptom appearance, 16 h after inoculation (Ct values = 28.29 ± 1.1). In olive samples, the derived melting curve was specific presenting a single dissociation peak (Tmelting = 88.7 °C). The designed assay was effectively applied in C. acutatum detection and quantification using infected olive samples, with a LOD of 0.59 ng and a LOQ of 1.8 ng, allowing its application to orchard management.

Astrocyte-mediated switch in spike timing-dependent plasticity during hippocampal development.

Presynaptic spike timing-dependent long-term depression (t-LTD) at hippocampal CA3-CA1 synapses is evident until the 3rd postnatal week in mice, disappearing during the 4th week. At more mature stages, we found that the protocol that induced t-LTD induced t-LTP. We characterized this form of t-LTP and the mechanisms involved in its induction, as well as that driving this switch from t-LTD to t-LTP. We found that this t-LTP is expressed presynaptically at CA3-CA1 synapses, as witnessed by coefficient of variation, number of failures, paired-pulse ratio and miniature responses analysis. Additionally, this form of presynaptic t-LTP does not require NMDARs but the activation of mGluRs and the entry of Ca2+ into the postsynaptic neuron through L-type voltage-dependent Ca2+ channels and the release of Ca2+ from intracellular stores. Nitric oxide is also required as a messenger from the postsynaptic neuron. Crucially, the release of adenosine and glutamate by astrocytes is required for t-LTP induction and for the switch from t-LTD to t-LTP. Thus, we have discovered a developmental switch of synaptic transmission from t-LTD to t-LTP at hippocampal CA3-CA1 synapses in which astrocytes play a central role and revealed a form of presynaptic LTP and the rules for its induction.