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- Are reusable dry electrodes an alternative to gelled electrodes for canine surface electromyography?Publication . Ribeiro, Ana M.; Brás, I.; Caldeira, L.; Peham, C.; Silva, H. Plácido da; Requicha, João F.Despite its increasing use in veterinary rehabilitation, practical constraints—such as skin preparation and single-use electrodes—limit the wider adoption of surface electromyography (sEMG). Having conventional pre-gelled Ag/AgCl electrodes as reference, we made a pioneering comparison of the performance of reusable soft polymeric dry electrodes for recording paraspinal muscle activity in dogs during treadmill walking. Twelve clinically healthy Dachshunds from both genders were evaluated under two conditions, namely: (i) dry electrodes on untrimmed hair; and (ii) pre-gelled electrodes after trichotomy. Signals were acquired from the longissimus dorsi muscle at 1 kHz, processed with standardized filtering and rectification, and analyzed in both time and frequency domains. Dry electrodes yielded higher amplitude and Root Mean Square (RMS) values, but slightly lower power spectral density metrics when compared to pre-gelled electrodes. Nevertheless, frequency-domain results were broadly comparable between configurations. Dry electrodes reduce the preparation time, avoid hair clipping, and allow reusability without major signal degradation. While pre-gelled electrodes may still offer marginally superior stability during movement, our results suggest that soft polymeric dry electrodes present a feasible, less invasive, and more sustainable alternative for canine sEMG. These findings support further validation of dry electrodes in clinical populations, particularly for neuromuscular assessment in intervertebral disk disease.
- Assessing environmental and pandemic influences on mortality through seasonal time series modelsPublication . Simões, João; Bernardo, Alexandra; Gonçalves, Luísa Lima; Brito, JoséUnderstanding the interplay between air pollution, climate variables, and mortality is essential for developing evidence-based public health policies and mitigating environmental health risks. Seasonal mortality patterns, particularly winter peaks, are often associated with increased air pollution and low temperatures. However, understanding the combined associations of these factors, especially under global disruptions such as the COVID-19 pandemic, poses a challenge for public health research and policy. This study shows that the monthly number of deaths from all-natural causes in Portugal from 2010 to 2022 exhibits a pronounced seasonal pattern, with winter mortality about 19 % higher than the annual mean. In the same period, air pollutant concentrations show clear seasonality, with NO2 (+24 %) and PM10 (+12 %) peaking in winter, while minimum temperatures fall about 42 % below the mean. Conversely, ozone exposure (SOMO35) reaches highest levels in summer, reflecting distinct seasonal burden. Using Seasonal Autoregressive Integrated Moving Average models with Predictor Variables (SARIMAX), the analysis identifies complex and lagged associations: PM10 exhibits a delayed effect at four months, while NO2 is associated with immediate increases on mortality. Temperature acts as a nonlinear and oscillatory predictor, with both acute effects and delayed cold-related risks over several months. The findings indicate that pandemic-related mortality disrupted typical seasonal patterns and hindered the detectability of subtler environmental effects. Comparing models that include or exclude COVID-19 mortality reveals clearer environmental associations in the non-COVID model, highlighting the value of distinguishing acute shocks from long-term conditions. Overall, the results underscore the need for stricter air quality standards during colder months and for adaptive public health strategies that consider cumulative and lagged environmental effects. While this study focused on adults aged 30 years and older, the findings suggest that older adults, individuals with chronic respiratory or cardiovascular conditions, and those in disadvantaged settings may be more susceptible to environmental stressors.
- Besnoitia besnoiti bradyzoite stages induce suicidal- and rapid vital-NETosisPublication . Zhou, Ershun; Silva, Liliana M. R.; Conejeros, Iván; Velásquez, Zahady D.; Hirz, Manuela; Gärtner, Ulrich; Jacquiet, Philippe; Taubert, Anja; Hermosilla, CarlosBesnoitia besnoiti is an obligate intracellular apicomplexan protozoan parasite, which causes bovine besnoitiosis. Recently increased emergence within Europe was responsible for significant economic losses in the cattle industry due to the significant reduction of productivity. However, still limited knowledge exists on interactions between B. besnoiti and host innate immune system. Here, B. besnoiti bradyzoites were successfully isolated from tissue cysts located in skin biopsies of a naturally infected animal, and we aimed to investigate for the first time reactions of polymorphonuclear neutrophils (PMN) exposed to these vital bradyzoites. Freshly isolated bovine PMN were confronted to B. besnoiti bradyzoites. Scanning electron microscopy (s.e.m.)- and immunofluorescence microscopy-analyses demonstrated fine extracellular networks released by exposed bovine PMN resembling suicidal NETosis. Classical NETosis components were confirmed via co-localization of extracellular DNA decorated with histone 3 (H3) and neutrophil elastase (NE). Live cell imaging by 3D holotomographic microscopy (Nanolive®) unveiled rapid vital NETosis against this parasite. A significant increase of autophagosomes visualized by specific-LC3B antibodies and confocal microscopy was observed in B. besnoiti-stimulated bovine PMN when compared to non-stimulated group. As such, a significant positive correlation (r = 0.37; P = 0.042) was found between B. besnoiti-triggered suicidal NETosis and autophagy. These findings suggest that vital- as well as suicidal-NETosis might play a role in early innate host defence mechanisms against released B. besnoiti bradyzoites from tissue cysts, and possibly hampering further parasitic replication. Our data generate first hints on autophagy being associated with B. besnoiti bradyzoite-induced suicidal NETosis and highlighting for first time occurrence of parasite-mediated vital NETosis.
- Besnoitia besnoiti tachyzoite replication in bovine primary endothelial cells relies on host Niemann–Pick type C protein 1 for cholesterol acquisitionPublication . Larrazabal, Camilo; Hermosilla, Carlos; Taubert, Anja; Silva, Liliana M. R.Besnoitia besnoiti is a cyst-forming apicomplexan parasite and the causal agent of bovine besnoitiosis. During early phase of infection, tachyzoites replicate within host endothelial cells in a host cell cholesterol-dependent process. By applying U18666A treatments, we here evaluated the role of Niemann–Pick type C protein 1 (NPC1) in both, intracellular B. besnoiti replication and host cellular cholesterol distribution. Additionally, B. besnoiti-driven changes in NPC1 gene transcription were studied by qPCR. Overall, U18666A treatments significantly reduced B. besnoiti proliferation and induced cholesterol accumulation in host cytoplasmic dense vesicles. However, NPC1 gene transcription was not affected by B. besnoiti infection.
- Black-and-White Ruffed Lemur (Varecia variegata) in captivity : analysis of the oral microbiota in a one health perspectivePublication . Silva, Carolina; Requicha, João F.; Martins, José J.; Duarte, Aida; Dias, Isabel R.; Viegas, Carlos A.; Saavedra, Maria J.This study aimed to characterize the susceptibility profile to antibiotics and biofilm formation of Gram-negative bacterial isolates obtained from the oral cavity of the black-and-white ruffed lemur (Varecia variegata). From eight individuals from a zoo located in Portugal, samples of the oral microbiota were collected with sterile swabs and then placed in closed tubes with a transport medium. Culture was carried out for media of Gram-negative bacteria. Twenty-two isolates were obtained and subjected to susceptibility tests to twenty-five antimicrobial agents belonging to seven different classes. All tested isolates demonstrated resistance to, at least, one antibiotic, and it was possible to observe multidrug resistance in 11 of the 22 isolates (50%). It should be noted that an isolate showed phenotypic resistance to imipenem, an antibiotic for exclusive use in a hospital environment. All the isolates showed an increasing ability of biofilm formation over time. The obtained results show that wild mammals in captivity could be reservoirs and potential sources of multi-resistant pathogens. In view of this fact and considering the One Health concept, it will be advisable to establish local monitoring programs worldwide that benefit and protect human, animal and environmental health.
- Bovine ocular squamous cell carcinoma : a descriptive epidemiological survey in the Azores, PortugalPublication . Bilhastre, Beatriz; Vala, Helena; Ribeiro, Ana Clara; Faria, Sara; Oliveira, Ana; Branco, Sandra; Pinto, CarlosBovine ocular squamous cell carcinoma (BOSCC) is the most common ocular tumour in cattle, with a multifactorial aetiology involving ultraviolet (UV) radiation, genetic factors, pigmentation, and management practices. A detailed epidemiological characterisation of BOSCC in the Azores, Portugal, is provided, with particular emphasis on its spatial distribution and potential risk determinants. Data were obtained through an epidemiological questionnaire completed by field veterinarians between August 2023 and March 2025. A total of 85 BOSCC cases were recorded across 62 farms—45 on Terceira Island and 17 on São Miguel Island. All affected animals were adult Holstein Friesian dairy cows, managed under extensive pasture-based systems. The nictitating membrane was the most frequently affected structure (69.5%), and multiple lesions occurred in 20% of the cases. Farms located at 200–400 m of altitude presented the highest number of cases. Continuous exposure to UV under pasture-based management represents the main environmental risk factor. Although periocular pigmentation may provide partial protection, other environmental and genetic factors can also contribute to tumour development. Artificial insemination is considered a promising preventive tool, enabling genetic selection for protective traits such as periocular pigmentation. This research provides the first regional epidemiological characterization of BOSCC in the Azores, highlighting the interplay among environmental, genetic, and management-related factors in disease occurrence.
- Breastfeeding as a strategic driver for One Health : a narrative reviewPublication . Machado, Vanessa; Regalo, Simone Cecílio Hallak; Ferreira, Luciano Maia Alves; Martinelli, Roberta Lopes de Castro; Trawitzki, Luciana Vitaliano Voi; Siéssere, Selma; Mendes, José João; Botelho, JoãoBreastfeeding is a renewable biological system that simultaneously advances human, environmental, and societal health. Human milk provides unparalleled nutrition and immunological protection, improving infant survival, neurodevelopment, and long-term metabolic outcomes, while reducing maternal risk of breast and ovarian cancer. However, and despite decades of evidence, only 48% of infants under six months are exclusively breastfed worldwide, and breastfeeding remains absent from most sustainability and One Health strategies. This narrative review synthesizes evidence demonstrating that breastfeeding functions as a low-carbon, zero-waste food system that avoids greenhouse gas emissions, land conversion, water consumption, and biodiversity loss linked to commercial milk formula production. At the societal level, breastfeeding reduces health-system costs, strengthens emergency resilience when supply chains fail, and generates long-term economic returns. By integrating evidence across human health, environmental impact and social determinants, this review positions breastfeeding as a strategic One Health intervention and a high-value investment for achieving multiple Sustainable Development Goals. Strengthening policy support—including protection against formula marketing, workplace accommodations, and expansion of baby-friendly systems—is essential to unlock breastfeeding’s potential for planetary and public health.
- BSF - time to change the fliesPublication . Menino, Regina; Murta, Daniel
- Building a Portuguese coalition for biodiversity genomicsPublication . Marques, João P.; Alves, Paulo C.; Amorim, Isabel R.; Lopes, Ricardo J.; Moura, Monica; Myers, Eugene; Sim-sim, Manuela; Sousa-Santos, Carla; Alves, M. Judite; Borges, Paulo A. V.; Brown, Thomas; Carneiro, Miguel; Carrapato, Carlos; Ceríaco, Luís M. P.; Ciofi, Cláudio; Silva, Luís P. da; Diedericks, Genevieve; Diroma, Maria Angela; Farelo, Liliana; Formenti, Giulio; Gil, Fátima; Grilo, Miguel; Iannucci, Alessio; Leitão, Henrique G.; Máguas, Cristina; Mc Cartney, Ann M.; Mendes, Sofia L.; Moreno, João M.; Morselli, Marco; Mouton, Alice; Natali, Chiara; Pereira, Fernando; Rego, Rúben M. C.; Resendes, Roberto; Roxo, Guilherme; Svardal, Hannes; Trindade, Helena; Vicente, Sara; Winkler, Sylke; Alvarenga, Marcela; Amaral, Andreia J.; Antunes, Agostinho; Campos, Paula F.; Canário, Adelino V. M.; Castilho, Rita; Castro, L. Filipe C.; Crottini, Angelica; Cunha, Mónica V.; Themudo, Gonçalo Espregueira; Esteves, Pedro J.; Faria, Rui; Fernandes, Carlos Rodríguez; Ledoux, Jean-Baptiste; Louro, Bruno; Magalhaes, Sara; Paulo, Octávio S.; Pearson, Gareth; Pimenta, João; Pina-Martins, Francisco; Santos, Teresa L.; Serrão, Ester; Melo-Ferreira, José; Sousa, Vítor C.The diverse physiography of the Portuguese land and marine territory, spanning from continental Europe to the Atlantic archipelagos, has made it an important repository of biodiversity throughout the Pleistocene glacial cycles, leading to a remarkable diversity of species and ecosystems. This rich biodiversity is under threat from anthropogenic drivers, such as climate change, invasive species, land use changes, overexploitation, or pathogen (re)emergence. The inventory, characterisation, and study of biodiversity at inter- and intra-specific levels using genomics is crucial to promote its preservation and recovery by informing biodiversity conservation policies, management measures, and research. The participation of researchers from Portuguese institutions in the European Reference Genome Atlas (ERGA) initiative and its pilot effort to generate reference genomes for European biodiversity has reinforced the establishment of Biogenome Portugal. This nascent institutional network will connect the national community of researchers in genomics. Here, we describe the Portuguese contribution to ERGA’s pilot effort, which will generate high-quality reference genomes of six species from Portugal that are endemic, iconic, and/or endangered and include plants, insects, and vertebrates (fish, birds, and mammals) from mainland Portugal or the Azores islands. In addition, we outline the objectives of Biogenome Portugal, which aims to (i) promote scientific collaboration, (ii) contribute to advanced training, (iii) stimulate the participation of institutions and researchers based in Portugal in international biodiversity genomics initiatives, and (iv) contribute to the transfer of knowledge to stakeholders and engaging the public to preserve biodiversity. This initiative will strengthen biodiversity genomics research in Portugal and fuel the genomic inventory of Portuguese eukaryotic species. Such efforts will be critical to the conservation of the country’s rich biodiversity and will contribute to ERGA’s goal of generating reference genomes for European species.
- Cattle effluents, either composted or bio-digested by Black Soldier Fly larvae, in the fertilization of ryegrass in sandy soilsPublication . Menino, Regina; Lopes, Ivã; Semedo, José N.; Scotti-Campos, Paula; Murta, Daniel; Castelo-Branco, Amélia; Nestler, Cecília; Moreira, OlgaThe relative agronomic performance of two organic fertilizers - effluents from intensive cattle farming, after composting either by traditional aerobic methods (compost) or by bio-digestion by larvae of the black soldier fly (Hermetia illucens, BSF) (frass) - in the growth of annual ryegrass (Lolium multiflorum L.) on a sandy soil was evaluated. For this purpose, a pot trial was conducted in a semi-controlled greenhouse. Eight treatments were defined, with four treatments using the compost and four treatments using frass (with a 10% nitroamoniacal nitrogen supplementation in all treatments), in addition to a negative control without fertilization, and a positive control with exclusively chemical nitroamoniacal fertilization. Regarding the biomass production, the BSF frass displayed a higher potential than the compost, with a significantly higher production from an N endowment equivalent to 280 kg ha-1 (252 kg organic N + 28 kg mineral N). While the yields obtained with compost were stable when 350 kg ha-1 (315 kg organic N + 35 kg mineral N) were provided, the yields obtained with BSF frass revealed a tendency to continue to progress with higher doses. This could be justified by the fact that the P2O5 and K2O content of BSF frass is significantly higher than that of the compost, and these nutrients continued to be mineralized over time. Furthermore, the obtained results confirm the potentiating effect of mineral fertilizers in the maximization of biomass production, when associated with organic fertilizers in adequate proportions. In addition, it was verified that the addition of frass enhanced the water use by plants, which was significantly higher than the treatments with compost, regardless of the N endowment. It was concluded that mixing chemical and organic fertilizers positively impacts soil fertility and the sustainability of the production as a whole, by increasing productivities in a sustainable way.