Heavy metal and metalloid concentrations in red deer (Cervus elaphus) and their human health implications from One Health perspective.

The red deer is an ungulate and large game species. The contamination of the ecosystems by metal(loid)s may lead to the exposure of animals (as well as humans) through water and food resources. The direct contact of hunters and wild animal meat consumers with deer carcasses may be a potential contaminant source. This study aimed to determine the metal(loid)s' concentrations in the liver and kidney of red deer from two regions of Portugal (Idanha-a-Nova and Lousã), and to relate these with histopathologic lesions. Thirteen young male deer were submitted to metal(loid) determination (As, Cd, Co, Cr, Cu, Pb, and Zn) by inductively coupled plasma mass spectrophotometry (ICP-MS) and histopathology examination. Renal Cd (8.072 ± 5.766 mg/kg dw) and hepatic Pb (3.824 ± 6.098 mg/kg dw) mean values were high, considering the maximum values for consumption established by the European Commission. The hepatic mean value of Cu was significantly higher in Idanha-a-Nova (150.059 ± 33.321 mg/kg dw), and it is at the Cu toxicity limit considered for ruminants (150 mg/kg). The pollution induced by Panasqueira mines (Castelo Branco) may be a possible explanation for some of the findings, especially the higher values of hepatic Cu and Pb found in Idanha-a-Nova deer. These results have high importance under a One Health perspective, since they have implications in public health, and pose at risk the imbalance of animal populations and ecosystems.

Leishmania Infection in Wild Lagomorphs and Domestic Dogs in North-East Spain.

Leishmania infantum is a zoonotic protozoan parasite distributed worldwide that is transmitted by phlebotomine sandflies. Dogs are the main reservoir for human infections. However, in recent years, the capacity of lagomorphs to contribute to Leishmania transmission has been confirmed. The present study aimed to assess Leishmania spp. exposure and infection in lagomorphs and sympatric domestic dogs in NE Spain. Sera from European hares, European rabbits, and rural dogs were tested for antibodies against L. infantum using an in-house indirect ELISA. PCR analysis targeting Leishmania spp. was performed in spleens from L. europaeus. Antibodies against Leishmania spp. were detected in all the species analyzed. Total sample prevalence was significantly higher in O. cuniculus (27.9%) than in L. europaeus (2.0%). Results of the PCR were all negative. The present study expands knowledge about Leishmania infections in free-ranging lagomorphs in the Iberian Peninsula, suggesting a more important role of O. cuniculus in the study area. Given the strong correlation between lagomorph densities and human leishmaniasis outbreaks in Spain, the high rabbit and human densities in NE Spain, and the high Leishmania spp. seroprevalence in rabbits, it becomes imperative to establish surveillance programs for lagomorphs in this region.

The first full study of heavy metal(loid)s in western-European hedgehogs (Erinaceus europaeus) from Portugal.

The western-European hedgehog (Erinaceus europaeus) is an insectivore with a wide distribution in Portugal and a potential tool for biomonitoring relevant One Health hazards, including heavy metal(loid)s' pollution. The aim of this study was to positively contribute to the current knowledge about the metal(loid) pollution in Portugal. Forty-six hedgehogs (from rescue centres; with known provenance) were necropsied. Sex, age category and weight were determined. Spines, liver and kidney were collected, and metalloid concentrations were determined by inductively coupled plasma mass spectrophotometry (ICP-MS). In general, results did not present alarming metal(loid) concentrations, with the exception of cadmium (Cd) (in the kidneys) and copper (Cu). Hedgehogs from Viana do Castelo and Viseu showed elevated concentrations of arsenic (As) and Castelo Branco presented concerning values of cadmium (Cd). Adult and heavier hedgehogs tended to present higher levels of metal(loid)s. Sex does not seem to significantly affect the metal(loid)s' concentrations. Further analysis would be needed to prioritize areas with detail and allow the application of the necessary mitigation strategies.

Heavy metals and metalloids in wild boars (Sus Scrofa) - a silent but serious public health hazard.

Wild boars (Sus scrofa) are part of the hunting economy and are highly consumed in the Iberian Peninsula, including in the Castile and Leon regions. As zoonotic diseases, chemical pollutants in wild boars' internal tissues should be interpreted as evidence of environmental contamination and a matter of concern for animal, human and ecosystem health; in other words, a One Health concern. Twenty-eight wild boars' livers and kidneys (n = 28) from Castile and Leon were submitted to metal(loid) determination (As, Cd, Co, Cr, Cu, Ni, Pb, and Zn) with inductively coupled plasma mass spectrophotometry (ICP-MS) and histopathological exam. Cd levels, especially in the kidneys (7.063 ± 7.271 mg/kg dw), were the most concerning results, considering the calculated maximum values for consumption (EC No. 915/2023) (2.491 mg/kg dw or 1.0 mg/kg ww). Wild boars with hydropic changes in the liver presented higher concentrations of Ni. Thus, the metal(loid) contamination of wild boar carcasses seems to be a "no trace" but very relevant problem that should raise awareness of a more accurate monitoring program and other strategies to avoid public health consequences.

A spontaneous ovarian teratoma in an FVB/n female mouse: Case report and literature review.

Background: Teratomas are rare types of germ cell neoplasms composed of various differentiated or undifferentiated tissues. Case Description: A 25-week-old female control FVB /n mouse in a 4-week toxicity study presented abdominal distension and poor body condition. It was euthanized, and the necropsy examination revealed a large mass connected to the tip of the right uterine horn, occupying the entire abdominal cavity. Microscopically, this mass showed areas of epidermal differentiation, with laminated keratin and sebaceous glands, differentiation into respiratory and digestive epithelium, cartilage, bone, and extensive areas of differentiation into the nervous tissue, being classified as an ovarian teratoma. Conclusion: As far as authors know, the occurrence of ovarian teratomas in the FVB/n mouse strain has never been previously described.

First morphological and molecular isolation of Talaromyces marneffei in beech marten (Martes foina) in Portugal.

Talaromyces marneffei is a zoonotic fungus that mostly infects immunocompromised individuals. For the first time, this fungus was isolated in an adult beech marten (Martes foina) hit by a car, found dead in Penamacor, Portugal. During the necropsy, different samples (skin, fur, lymph nodes, lung, spleen, kidneys, and brain) were collected and processed for microbiology (including mycology) and molecular biology. T. marneffei was identified through its mycological characteristics and confirmed by PCR in hair samples. No other lesions or alterations were reported, except a concomitant presence of M. avium subsp. paratuberculosis in lung, kidney and brain samples. To the authors' knowledge, this is the first description of this fungus beech marten, as well as the first case of co-infection with M. avium subsp. paratuberculosis in wildlife fauna. These results suggest a sylvatic life-cycle of T. marneffei, involving beech martens, in Portugal.

High Levels of Heavy Metal(loid)s Related to Biliary Hyperplasia in Hedgehogs (Erinaceus europaeus).

Heavy metal(loid) pollution of ecosystems is a current One Health problem. The liver is one of the most affected organs in cases of acute or chronic exposure to abnormal amounts of these substances, inducing histopathologic lesions. In order to assess the influence of heavy metal(loids), forty-five European hedgehogs (Erinaceus europaeus) were submitted to necropsy, and liver samples were collected for a routine histopathology exam and metal(loid)s determination (As, Cd, Co, Cr, Cu and Pb) by ICP-MS. Age was estimated during the necropsy exam. Biliary hyperplasia was the most frequent lesion observed (16/45; 35.56%). No statistically significant associations were found between biliary hyperplasia and age or sex. Metal(loid)s' concentrations were higher in animals with biliary hyperplasia (except for As). There was a statistically significant difference for both Cd and Co. For As, Cd and Co, cubs and juveniles animals showed significantly lower concentrations than elder individuals. Only for Pb were significant differences found between females and males. As described in the literature, exposure to metal(loid)s may be a cause of biliary hyperplasia, although further research (including the use of biochemical methods) is needed to support these results. To the authors' knowledge, this is the first report of this association in hedgehogs.

Talaromyces marneffei isolated from nasal swabs in Portugal.

Mycobiota are essential to the health of any living being, creating a balanced and complex interaction between bacteria, the immune system, and the tissue cells of the host. Talaromyces marneffei (also known as Penicillium marneffei) is a dimorphic fungus, endemic in South Asia, which often causes a life-threatening systemic fungal infection (called penicilliosis), particularly in immunocompromised hosts. Nasal swabs from 73 healthy volunteers were analysed to characterize their mycobiota, through its cultural characteristics, morphology, and molecular methods (PCR). All volunteers were also asked to answer to an anonymous questionnaire. Three women were positive (and asymptomatic) for T. marneffei. One of them was reported to have lupus. This study contributes to improving our knowledge about human normal mycobiota, identifying mycotic agents that may cause complicated systemic infections (as T. marneffei), especially in immunosuppressed patients, as well as other possible risk factors of exposure or prognosis.

• Talaromyces marneffei is a zoonotic fungus that may be responsible for life-threatening systemic infections in immune-comprised patients. • Talaromyces marneffei was identified in nasal swabs from asymptomatic volunteers. • This suggests that this fungus may be part of the nasal normal mycobiota of some humans.

Do Urban Hedgehogs (Erinaceus europaeus) Represent a Relevant Source of Zoonotic Diseases?

Urban fauna is defined as animal species that can live in urban environments. Several species, including the western-European hedgehog (Erinaceus europaeus), have now been identified as part of this urban fauna, becoming permanent residents of parks and gardens in different cities across Europe. Due to the importance that this phenomenon represents for zoonotic disease surveillance, several authors have been conducting zoonotic agents' surveys on hedgehog. The aim of this study is to compare zoonotic diseases' prevalence in hedgehogs in urban environments with those from more rural areas. A systematic review with meta-analysis of twelve studied of zoonotic diseases' (in urban and rural areas of Europe) was therefore conducted for this purpose. Fifteen different zoonoses have been assessed in urban environments and six in rural areas. Anaplasma phagocytophilum was the most prevalent zoonotic agent found in urban habitats (96%). Dermatophytosis shows statistically significant differences between locations (p-value < 0.001), with a higher prevalence in urban Poland (55%; n = 182). Our results suggest further research and a standardized monitoring of different hedgehog populations are essential to understanding the epidemiology of several zoonotic pathogens in different habitat types (urban, rural, natural, industrial, etc.) and preventing possible disease outbreaks.

Practitioners' knowledge, acceptability and use of external memory aids with individuals with cognitive deficits: An exploratory study.

External memory aids (EMA) are within the most effective cognitive rehabilitation techniques, having demonstrated a positive impact in terms of memory functioning in individuals with multiple cognitive deficits. Despite its proven efficacy, there is yet poor dissemination of these techniques in clinical settings. The current study aims to evaluate the level of knowledge, degree of use, and usage expectations of EMAs by health practitioners, responsible to implement these techniques. A quantitative, descriptive, and cross-sectional study was developed, and 120 practitioners working with cognitively impaired patients participated in the study. One questionnaire was developed to assess participants' knowledge and use of EMAs. Results indicate that the level of global knowledge regarding EMAs is poor, despite superior to its actual use. The degree of perceived acceptance of these prosthetics by their users is positive and stronger than the likelihood of practitioners to use these tools with their patients. This study suggests that the actual implementation of EMAs as rehabilitation tools is still poor, despite the growing evidence-based research highlighting its effectiveness to compensate for cognitive deficits. Future studies should target the current factors that are influencing the underutilization of EMAs, to improve and optimize its dissemination and to benefit cognitively impaired patients.

Evolution of Models of Prostate Cancer: Their Contribution to Current Therapies.

Prostate cancer (PCa) is among the most frequent cancers worldwide. Nowadays, several therapeutic strategies are available for PCa treatment, namely chemotherapy, immunotherapy, radiotherapy, and hormonal therapy. Despite existing therapeutic approaches, in vitro and in vivo models are essential to better understand cancer development and search for more effective therapies, with a positive impact in cancer patient survival and quality of life. Among several models available, the rat model is the one most frequently used, since it shares anatomical, physiological, pathological, and behavioral features with humans. Animal models can be classified as: spontaneous, chemically-induced; hormonally-induced; implantation of cancer cell lines obtained from humans or from the same species, in the place of disease development or in a different place; and genetically-modified models. The chemically-induced models are among the most frequently used for PCa research. This manuscript provides a comprehensive overview of PCa models, presenting their application, advantages, and disadvantages, and their importance for the development of current therapies for prostate cancer.

Wild Boars (Sus scrofa, L. 1758) from Castile and Leon Region (Spain): A Histopathology Survey.

Wild boars are wild ungulates with a wide distribution in Europe, with a relevant role in wildlife and public health. In Spain, high (and sometimes artificial) densities of wild boars are responsible for several health problems. Regular surveys, with hunters' collaboration, are crucial to monitor these diseases. Histological analyses were performed for lung, liver, and kidneys from 72 wild boars (58 from Zamora, 16 from Palencia). Lungs were the most affected organs, mainly revealing parasitic pneumonia (34.7%). Hydropic, vacuolar, and other cellular changes (33.3%) and congestion (16.7%) were found in the liver, and only 30.6% of the wild boars presented no alterations in this organ. Regarding the kidney, non-purulent nephritis (22.2%) was the most common lesion. This study gives an overview of the health status of wild boar populations in Castile and León. Other laboratory analyses are needed to obtain definitive diagnoses of these lesions, reach other conclusions, or apply any mitigation strategies to protect animals' or consumers' health.

3D-Printed Teeth in Endodontics: Why, How, Problems and Future-A Narrative Review.

Three-dimensional printing offers possibilities for the development of new models in endodontics. Numerous studies have used 3D-printed teeth; however, protocols for the standardization of studies still need to be developed. Another problem with 3D-printed teeth is the different areas of literature requested to understand the processes. This review aims to gather evidence about 3D-printed teeth on the following aspects: (1) why they are advantageous; (2) how they are manufactured; (3) problems they present; and (4) future research topics. Natural teeth are still the standard practice in ex vivo studies and pre-clinical courses, but they have several drawbacks. Printed teeth may overcome all limitations of natural teeth. Printing technology relies on 3D data and post-processing tools to form a 3D model, ultimately generating a prototype using 3D printers. The major concerns with 3D-printed teeth are the resin hardness and printing accuracy of the canal anatomy. Guidance is presented for future studies to solve the problems of 3D-printed teeth and develop well-established protocols, for the standardization of methods to be achieved. In the future, 3D-printed teeth have the possibility to become the gold standard in ex vivo studies and endodontic training.

Effects of testosterone and exercise training on bone microstructure of rats.

Background and Aim: Male hypogonadism results from failure to produce physiological levels of testosterone. Testosterone in men is essential in masculine development, sperm production, and adult man's health. Osteoporosis is one of the consequences of hypogonadism. Regular physical exercise and exogenous testosterone administration are frequently used to prevent or treat this condition. This study aimed to understand the effects of lifelong exercise training and testosterone levels (isolated and together) in the main bone structure parameters. Materials and Methods: A total of 24 rats were used and randomly divided into four groups: Control group (CG; n=6), exercised group (EG, n=6), testosterone group (TG, n=6), and testosterone EG (TEG, n=6). A micro-computed tomography equipment was used to evaluate 15 bone parameters. Results: Both factors (exercise training and testosterone) seem to improve the bone resistance and microstructure, although in different bone characteristics. Testosterone influenced trabecular structure parameters, namely, connectivity density, trabecular number, and trabecular space. The exercise promoted alterations in bone structure as well, although, in most cases, in different bone structure parameters as bone mineral density and medullar mineral density. Conclusion: Overall, exercise and testosterone therapy seems to have a synergistic contribution to the general bone structure and resistance. Further studies are warranted, comparing different individual factors, as gender, lifestyle, or testosterone protocols, to constantly improve the medical management of hypogonadism (and osteoporosis).

Biomonitoring metals and metalloids in wild mammals: invasive versus non-invasive sampling.

Heavy metal and metalloid pollution is a matter of concern in animal, human and environmental health (One Health) and also in wildlife conservation worldwide. Studying wild mammals in toxicology has been contributing significantly to our knowledge, namely to find out the most critical regions, to understand bioaccumulation and biomagnification phenomena or to evaluate their toxic effects. However, not all the animal tissues and organs provide the same information or should be interpreted in the same way. The best sample to use will depend on the objectives and conditions of the study. This review aims to compare invasive and non-invasive samples to biomonitor heavy metals, providing a brief resume of their advantages, limitations and examples of use. Further research, using a wider range of mammalian species, is required to establish what information can be obtained in biomonitoring studies that use non-invasive samples (such as hair, faeces and parasites) and/or invasive samples (such as blood, liver, kidney, bone and other organs).

Animal Models in Pharmacology: A Brief History Awarding the Nobel Prizes for Physiology or Medicine.

BACKGROUND: The Nobel Prize of Physiology or Medicine (NPPM) has recognized the work of 222 scientists from different nationalities, from 1901 until 2020. From the total, 186 award researchers used animal models in their projects, and 21 were attributed to scientists and projects directly related to Pharmacology. In the most recent years, genetics is a dominant scientific area, while at the beginning of the 20th century, most of the studies were more related to anatomy, cytology, and physiology. SUMMARY: Mammalian models were used in 144 NPPM projects, being rodents the most used group of species. Moreover, 92 researchers included domestic species in their work. The criteria used to choose the species, the number of animals used and the experimental protocol is always debatable and dependent on the scientific area of the study; however, the 3R's principle can be applied to most scientific fields. Independently of the species, the animal model can be classified in different types and criteria, depending on their ecology, genetics, and mode of action. Key-Messages: The use of animal models in NPPM awarded projects, namely in Pharmacology, illustrates their importance, need and benefit to improve scientific knowledge and create solutions. In the future, with the contribute of technology, it might be possible to refine the use of animal models in pharmacology studies.

Gastrointestinal Parasitism in Przewalski Horses (Equus ferus przewalskii).

PURPOSE: Przewalski's horse (Equus ferus przewalskii) is a subspecies of Equus ferus classified as endangered by the International Union for Conservation of Nature (IUCN), whose gastrointestinal (GI) parasitism has been studied in different populations, in situ and ex situ, during the last decades. We aimed to summarize the information available on the subject. METHODS: In this review, we collected information from distinct published literature, interpreted it, compared it and related the Przewalski's horse biology and ecology with different findings. RESULTS: Many similarities can be found between the Przewalski's horse populations and the domestic horses regarding their gastrointestinal parasite community. However, not all the GI parasites described in the domestic horse have been reported in Przewalski's horse's surveys. The most prevalent parasites found in Przewalski's horses belong to the Strongylidae family, namely the Cyathostominae. CONCLUSIONS: More continuous monitoring studies should be carried on in different in situ and ex situ locations to consistently support the one-off surveys. It might be necessary to better correlate the parasitological findings of this subspecies with other animals that share the same habitat and ecosystem, for instance, intermediate and reservoir hosts, from different animal taxonomic groups.