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Based on previous evidence demonstrating the efficacy of inactivated mycobacteria for the control of fish mycobacteriosis, we explored the protective efficacy of two inactivated Mycobacterium bovis administered via parenteral and mucosal routes against Mycobacterium marinum infection mimicking natural conditions in the zebrafish model of tuberculosis. Although we did not observe a clear effect of any of the immunostimulants on mycobacterial burden, the results showed a significant increase in TLR2 and TLR4 gene expression levels in fishes parenterally immunized with inactivated Bacillus Calmette-Guérin (BCG). Our findings demonstrated that the TLR2 and the TLR4 signaling pathways are involved in the immune response elicited by inactivated mycobacteria in the zebrafish model of tuberculosis and support the use of inactivated mycobacteria in vaccine formulations for the control of mycobacteriosis.

[ES] En este trabajo abordamos de manera monográfica el contexto arqueológico de la tumba 45 de la necrópolis ibérica III de Alarcos, actualmente en proceso de excavación y de estudio. La tumba, adscrita a un individuo adulto posiblemente masculino, presenta como parte del ajuar un thymiaterion cerámico inédito compuesto por dos piezas: un soporte calado en su parte inferior y una cazoleta con asa y base perforada. Los paralelos formales que presenta con diferentes soportes calados del oppidum oretano del Cerro de las Cabezas (Valdepeñas, Ciudad Real) y la relación que mantiene la tumba 45 con otros contextos funerarios cercanos permiten datar la tumba entre los siglos IV y III a. C. En este estudio rastreamos los diferentes paralelos de las dos piezas que componen el thymiaterion cerámico en el contexto de la Oretania, así como los thymiateria de bronce y los pebeteros de terracota, y contrastamos si pueden clasificarse o no como posibles quemadores de perfumes. Por otro lado, analizamos el registro arqueológico de los yacimientos oretanos para documentar el empleo del perfume en la zona de estudio, analizando las diferentes tipologías de ungüentarios cerámicos, pasta vítrea y fayenza. Así, hemos podido atestiguar que, desde el siglo VI a. C. hasta época iberorromana, en la mayor parte de los casos el perfume aparece asociado a un uso cultual o funerario del mismo. Finalmente, un análisis detallado del registro material permite relacionar la combustión y el uso de aceites perfumados con el culto a una divinidad femenina relacionada con la fertilidad, la abundancia y la regeneración del ciclo vital., [EN] In this work we monographically address the archaeological context of tomb 45 in the Iberian necropolis III of Alarcos, currently under excavation and study. The tomb, attributed to a possibly male adult individual, features, as part of its burial offerings, an unpublished ceramic thymiaterion composed of two pieces: a perforated base stand and a cup with a handle and perforated base. The formal parallels it exhibits with different perforated stands from the Oretanian oppidum of Cerro de las Cabezas (Valdepeñas, Ciudad Real), and its relationship with nearby funerary contexts, allow dating the tomb to the 4th to 3rd centuries BCE. In this study, we trace various parallels for the two pieces that make up the ceramic thymiaterion in the Oretanian context, as well as bronze thymiateria and terracotta censers, and assess whether they can be classified as possible perfume burners. Furthermore, we analyze the archaeological record of Oretanian sites to document the use of perfume in the study area, examining different typologies of ceramic unguentaria, glass paste, and faience. Thus, we have been able to ascertain that, from the 6th century BCE to the Iberian-Roman period, in most cases, perfume is associated with a cultic or funerary use. Finally, a detailed analysis of the material record allows us to link the combustion and use of perfumed oils to the worship of a female deity associated with fertility, abundance, and the regeneration of the life cycle.

Available methods to census exophilic tick populations have limitations in estimating true population size due to their inability to capture a high proportion of the actual tick population. We currently ignore the efficacy of these methods to capture questing Hyalomma spp. ticks, vectors of the Crimean-Congo hemorrhagic fever virus. To address the need of accurately estimating questing densities of Hyalomma spp., we designed a field experiment to test the efficacy of blanket dragging, blanket flagging, CO2-baited traps, and an ad hoc designed method, absolute surface counts, in capturing adult Hyalomma lusitanicum ticks from known numbers of preset fluorescent-marked ticks. The experiment was designed in 2 stages to estimate the point (1-day sampling) and cumulative (3-day serial sampling) efficacy of the methods under varying sampling effort and habitat. Tick survival, host interference, and weather effects on efficacy were controlled for in multiple regression models. There was high variability in method efficacy for capturing ticks, which was also modulated by effort, habitat, tick density, hosts, and soil temperature. The most effective method was absolute surface counts for both point estimates (39%) and cumulative efficacy (83%). CO2-baited traps reached a maximum efficacy of 37%, while blanket dragging and blanket flagging captured a maximum of the 8% of the marked ticks. Our results reveal the strengths and weaknesses of the different tick capture methods applied to adult H. lusitanicum and lay the groundwork for more accurate inferences about the true size of exophilic tick populations.

Crimean-Congo haemorrhagic fever (CCHF) virus (CCHFV) is a tick-borne zoonotic pathogen that can cause a lethal haemorrhagic disease in humans. Although the virus appears to be endemically established in the Iberian Peninsula, CCHF is an emerging disease in Spain. Clinical signs of CCHFV infection are mainly manifested in humans, but the virus replicates in several animal species. Understanding the determinants of CCHFV exposure risk from animal models is essential to predicting high-risk exposure hotspots for public health action. With this objective in mind, we designed a cross-sectional study of Eurasian wild boar (Sus scrofa) in Spain and Portugal. The study analysed 5,291 sera collected between 2006 and 2022 from 90 wild boar populations with a specific double-antigen ELISA to estimate CCHFV serum prevalence and identify the main determinants of exposure probability. To do so, we statistically modelled exposure risk with host- and environment-related predictors and spatially projected it at a 10 × 10 km square resolution at the scale of the Iberian Peninsula to map foci of infection risk. Fifty-seven (63.3 %) of the 90 populations had at least one seropositive animal, with seroprevalence ranging from 0.0 to 88.2 %. Anti-CCHFV antibodies were found in 1,026 of 5,291 wild boar (19.4 %; 95 % confidence interval: 18.3–20.5 %), with highest exposure rates in southwestern Iberia. The most relevant predictors of virus exposure risk were wild boar abundance, local rainfall regime, shrub cover, winter air temperature and soil temperature variation. The spatial projection of the best-fit model identified high-risk foci as occurring in most of western and southwestern Iberia and identified recently confirmed risk foci in eastern Spain. The results of the study demonstrate that serological surveys of CCHFV vector hosts are a powerful, robust and highly informative tool for public health authorities to take action to prevent human cases of CCHF in enzootic and emergency set

To estimate the determinants of spatial variation in Crimean-Congo hemorrhagic fever virus (CCHFV) transmission and to create a risk map as a preventive public health tool, we designed a survey of small domestic ruminants in Andalusia, Spain. To assess CCHFV exposure spatial distribution, we analyzed serum from 2,440 sheep and goats by using a double-antigen ELISA and modeled exposure probability with environmental predictors by using generalized linear mixed models. CCHFV antibodies detected in 84 samples confirmed low CCHFV prevalence in small domestic ruminants in the region. The best-fitted statistical model indicated that the most significant predictors of virus exposure risk were cattle/horse density and the normalized difference vegetation index. Model validation showed 99.7% specificity and 10.2% sensitivity for identifying CCHFV circulation areas. To map CCHFV exposure risk, we projected the model at a 1 × 1-km spatial resolution. Our study provides insight into CCHFV ecology that is useful for preventing virus transmission.

Telemetry using Global Positioning Systems (GPS) and camera trapping are two widely used approaches to detect animal interactions involving wildlife. However, while the former follows an individual-based approach, the latter follows a population-based approach, which may lead to different outcomes when describing interaction networks. In this study, we compared the performance of GPS telemetry, regular grid camera trapping (CT-RG), and resource monitoring camera trapping (in which cameras are placed monitoring a valuable resource for animals, such as water or food; CT-RM) in detecting and quantifying animal interactions. We used spatially explicit simulated trajectories of 150 individuals of 3 imaginary species. We simulated 9 different scenarios of resource visiting frequency and spatial resource aggregation in the environment to assess the potential influence of animal behavior and landscape structure. The simulated trajectories were sampled using the three approaches with six different sampling efforts each, and the direct and indirect interactions between the three species were quantified. Additionally, CT-RG and CT-RM were applied in Doñana National Park to evaluate their differences and limitations in field application. In terms of simulation, CT-RM offered the best performance in detecting direct interactions, and GPS achieved good performance when using a high number of fixes per hour. Both CT-RM and GPS also achieved good performance in detecting indirect interactions. CT-RG, on the other hand, missed most direct and indirect interactions, but correctly identified the pairs of species that interacted indirectly at some point. In the field trial, CT-RM and CT-RG detected almost the same number of species pairs interacting indirectly. However, CT-RG missed at least half of the species pairs that interacted directly. The spatial aggregation of the resource favors the performance of GPS over CT-RM, while the opposite occurs when the frequency of animal visits t

In the field of assisted reproductive technologies, oocyte in vitro maturation (IVM) has emerged as a potential alternative to overcome the limitations associated with standard ovarian stimulation treatments. However, current IVM protocols lack standardization and yield oocytes of inferior quality compared to those matured in vivo. To address this issue, novel biomaterials like hydrogels offer distinct advantages in cell culture by providing a 3D cellular environment and facilitating easy adjustment and characterization of mechanical properties, such as hydrogel stiffness. Herein, a novel and reusable bilayer hydrogel system is presented that accurately mimics the mechanical properties of the microenvironment surrounding oocyte maturation. The system comprises an outer layer fabricated from either a 3D-printed synthetic polymer (2-vinyl-4,6-diamino-1,3,5-triazine, (VDT)) or a natural polymer (chitosan), along with an inner layer composed of alginate. By faithfully replicating the mechanical properties of native tissue within a 3D culture environment, this system significantly improves the quality and developmental capacity of oocytes, thus resulting in successful embryo development. Overall, this innovative system holds great potential for future medical research and applications in cell culture, thus representing a significant advancement in assisted reproductive technologies.

In dry climates, livestock farming contributes to waterhole creation and maintenance, thereby contributing to biodiversity conservation. However, these lentic water bodies also represent a critical environmental connection between microorganisms and their vertebrate hosts since the water can facilitate pathogen persistence and transmission. Therefore, interventions for tuberculosis (TB) risk mitigation at the wildlife-livestock interface often focus on segregating host species at water points. We hypothesized that waterhole characteristics modulate their use by vertebrates and subsequent pathogen exposure risk. We visited 298 waterholes on 80 TB-positive and 40 TB-negative cattle farms in Spain to assess differences in waterhole characteristics and identify possible management implications. There was an average of 2.7 waterholes per farm. This represents 0.02 waterholes per km2 of farmland and 3.5 m2 of lentic waterbodies per km2 of farmland. Among the studied waterholes, 95% were man-made. Waterholes on TB-positive farms were 42% closer to covering vegetation than waterholes located on TB-negative ones. Farms with man-made waterholes showed a higher risk of TB than those with natural ones. The density of waterholes per surface unit was negatively associated with the farm TB risk. Waterholes placed on TB-positive farms were more intensively trampled by livestock. The best model explained farm positivity to TB as a function of the distance from the waterholes to the nearest cover vegetation, the aquatic vegetation richness found in the waterhole, the intensity of livestock use (trampling), the surrounding waterhole density, and the interaction between waterhole perimeter and the total number of signs of potential wildlife TB hosts per waterhole. Identifying the key waterhole features related to infection risk might allow designing One Health−inspired biosecurity measures such as increasing the number of waterholes, placing new waterholes farther away from cover, or f

Epizootic hemorrhagic disease (EHD) virus serotype 8 (EHDV-8) emerged in Spain in autumn 2022. In this study, we aimed to (1) characterize the clinical and lesional presentation of EHDV infection in European red deer (Cervus elaphus), and (2) study the spatial spread of the virus in wild ruminants in Spain after its introduction, in 2022/2023. We confirmed EHDV infection in two clinically compatible sick red deer by PCR and detection of anti-EHDV specific antibodies. EHDV infection occurred in red deer with hyperacute to acute clinical signs and lesions associated to vascular changes leading to death of the animals. Partial sequences of variable segment 2 (VP2) and segment 5 (NS1) genes of the detected viruses had >99% nucleotide identity with EHDV-8 sequences from Tunisia and Italy. In a cross-sectional serological study of EHDV in 592 wild ruminants, mainly red deer (n=578), in southwestern Spain, we detected anti-EHDV antibodies in 37 of 592 samples (6.3%; 95% confidence interval: 4.3–8.2), all from red deer and from the localities where clinical cases of EHD were confirmed in red deer. We conclude that EHDV-8 infection causes severe EHD in European red deer. The serosurvey revealed a limited spread of EHDV-8 in Spanish wild ruminant populations in the first year of virus detection in Spain.

[EN] Atrial fibrillation (AF) is a prevalent cardiac arrhythmia often treated concomitantly with other cardiac interventions through the Cox-Maze procedure. This highly invasive intervention is still linked to a long-term recurrence rate of approximately 35% in permanent AF patients. The aim of this study is to preoperatively predict long-term AF recurrence post-surgery through the analysis of atrial activity (AA) organization from non-invasive electrocardiographic (ECG) recordings. A dataset comprising ECGs from 53 patients with permanent AF who had undergone Cox-Maze concomitant surgery was analyzed. The AA was extracted from the lead V1 of these recordings and then characterized using novel predictors, such as the mean and standard deviation of the relative wavelet energy (RWEm and RWEs) across different scales, and an entropy-based metric that computes the stationary wavelet entropy variability (SWEnV). The individual predictors exhibited limited predictive capabilities to anticipate the outcome of the procedure, with the SWEnV yielding a classification accuracy (Acc) of 68.07%. However, the assessment of the RWEs for the seventh scale (RWEs7), which encompassed frequencies associated with the AA, stood out as the most promising individual predictor, with sensitivity (Se) and specificity (Sp) values of 80.83% and 67.09%, respectively, and an Acc of almost 75%. Diverse multivariate decision tree-based models were constructed for prediction, giving priority to simplicity in the interpretation of the forecasting methodology. In fact, the combination of the SWEnV and RWEs7 consistently outperformed the individual predictors and excelled in predicting post-surgery outcomes one year after the Cox-Maze procedure, with Se, Sp, and Acc values of approximately 80%, thus surpassing the results of previous studies based on anatomical predictors associated with atrial function or clinical data. These findings emphasize the crucial role of preoperative patient-specific ECG si

Addressing large bone segmental defects remains a profound challenge in orthopaedic surgery, demanding graft designs that mirror patient-specific defects while ensuring vascularization, osteoconduction, and structural integrity. Conventional approaches fall short in achieving this level of customization and functionality. This work leverages fuse filament fabrication (FFF) to fabricate innovative yttria stabilize zirconia graft designs that combine several strategies to tackle these multifaceted challenges. Graft designs ingeniously incorporate dense and porous structures mimicking the intricate architecture of natural bone for improved structural integrity and different ridges and protrusions to enhance the fixation. Additionally, these grafts undergo infiltration with PCL/PVA/AgNPs composite polymers, enriching biocompatibility, structural integrity, and antibacterial properties, fostering conducive conditions for optimal bone growth. Attaining printable filaments with YSZ powders below 50 nm and loads reaching 50 vol% marks a pioneering advancement in bone graft fabrication via Fused Filament Fabrication (FFF). This enabled high densification and low grain size at reduced sintering temperatures, imperative to achieve high mechanical performance. The biodegradation capacity of biopolymers in PBS and their role in the mechanical behaviour of grafts have been assessed, demonstrating the beneficial potential of the multicomponent graft proposed reaching strength values of 90–105 MPa.

The behaviour of a parallel-plate capacitor connected to a sinusoidal alternating voltage is analysed and the electric and magnetic fields, as well as the electric and magnetic energies inside the capacitor are obtained. Aspects related to Maxwell's equations, the Poynting vector and the conservation of electromagnetic energy are also discussed. Under certain conditions the electromagnetic energy stored in the capacitor remains constant and the behaviour of the capacitor at high frequencies is like that of an oscillating L-C circuit consisting of an inductor and a capacitor. The general situation is considered, valid for both low and high frequencies, and what is discussed here is of interest to students of an undergraduate electrodynamics course.