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This study aims to investigate the meteorological variables determining Cupressaceae pollen grain disruption in the environment. A parallel sampling of pollen grains and disrupted Cupressaceae pollen grains was performed in six cities using two Spanish aerobiological networks. The pollen concentrations, disrupted pollen concentrations, percentage of disrupted pollen and number of days when the percentage of disrupted pollen was above or equal to 50 % were quantified during two pollen seasons. The concentrations were determined following the standardised method EN 16868. Results show that the concentrations of pollen grains and disrupted pollen grains were not determined by geographical features and rarely by bioclimatic variables or indexes but by the ornamental use of the specimens in the vicinity of the pollen sampler, highlighting the possibility of using management practices to reduce exposure to allergens in the cities. African dust outbreaks coincided with higher concentrations of pollen grains and disrupted pollen grains, but the reduced percentage of disrupted pollen grains pointed to a non-causal relationship with long-distance transport. The effect of wind and maximum gusts remained negligible. The triggering factor for pollen disruption was the amount of water in the atmosphere, mainly reported as relative humidity. Rainfall increased the effect of disruption due to pollen grain swelling caused by its wash-out effect. The higher the relative humidity, the higher the disrupted pollen concentrations. This aligns with the mechanism of Cupressaceae reproduction since the family needs a water medium in the form of pollination droplets for the pollination tube to develop and the pollen grain to perform its biological function. Therefore, people that develop allergic symptoms to Cupresaceae pollen should avoid exposure during days with high relative humidity in the main pollen season.

[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

We measure the magnetic field produced by a linear octupole formed by four magnets placed in such a way that both their magnetic moments and their quadrupole moments cancel each other. The magnetic field is measured with the magnetic sensor of a smartphone together with a suitable app, using the Android operating system. The main objective is to determine the dependence of the x-component of the magnetic field on distance and compare the experimental results with the theoretical result that tells us that this dependence is a quintic decrease. Modern smartphones are equipped with various sensors: accelerometer, gyroscope, proximity sensor, ambient light sensor, capacitive sensors, magnetic sensor, GPS, and maybe others. The magnetic sensor allows us to measure the magnetic field and works by making use of the classical Hall effect. This magnetic sensor is being used in introductory physics laboratories in interesting ways.

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.

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.

Telomere length and dynamics are commonly used biomarkers of somatic state, yet the role of telomeres underlying the aging process is still debated. Indeed, to date, empirical evidence for an association between age and telomere length is mixed. Here, we test if the age-dependency of the association between age and telomere length can provide a potential explanation for the reported inconsistencies across studies. To this end, we quantified telomere length by telomere restriction fragment analysis in two groups of Japanese quail (Coturnix japonica) that differed in their age distribution. One group consisted of young adults only, whereas the second group consisted of adults across a wide range of ages. In the young adults group, there was a highly significant negative association between telomere length and age, whereas no association between age and telomere length was found in the all-ages adults group. This difference between groups was not due to telomere length-dependent selective disappearance. Our results shows that the association between telomere length and age is age-dependent and suggest that the costs and benefits associated with telomere maintenance are dynamic across an individual's life course.

The number and quality of social bonds can have major consequences for fitness. For example, in socially monogamous species with biparental care, pair bond quality has been linked to the latency to breed as well as the number and survival of offspring. Given these benefits, what mechanisms prevent some individuals from forming strong pair bonds? Markers of physiological stress and ageing, such as oxidative stress and telomere length, might mediate individual differences in behavioural performance. However, the possibility that physiological stress could also constrain the strength of the pair bond has rarely been investigated. We show that in captive colonies of the socially monogamous, gregarious zebra finch, Taeniopygia guttata, individuals with higher levels of plasma lipid oxidative damage formed weaker pair bonds. This effect was sex specific: while males with more oxidative damage spent less time in bodily contact with their prospective breeding partners, no such link was found in females. Although females experienced higher absolute levels of plasma oxidative damage, pair bond investment may have been more constrained in males due to the costly expression of their sexually selected traits. Pair bond strength was not associated with levels of the key antioxidant glutathione or with telomere length. Individuals’ ability to form strong pair bonds may thus be constrained by their levels of oxidative damage, with potential downstream effects on fitness.

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.

Acetonitrile (CHCN) is present in the interstellar medium (ISM) in a variety of environments. However, at the ultracold temperatures of the ISM, radical-molecule reactions are not widely investigated because of the experimental handicap of getting organic molecules in the gas phase by conventional techniques. The CRESU (French acronym for Reaction Kinetics in a Uniform Supersonic Flow) technique solves this problem. For this reason, we present in this work the kinetic study of the gas-phase reaction of CHCN with one of the most ubiquitous radicals, the hydroxyl (OH) radical, as a function of temperature (11.7-177.5 K). The kinetic technique employed to investigate the CHCN + OH reaction was the pulsed laser photolysis-laser induced fluorescence. The rate coefficient for this reaction k(T) has been observed to drastically increase from 177.5 K to 107.0 K (about 2 orders of magnitude), while the increase in k(T) from 107.0 K to 11.7 K was milder (around 4 times). The temperature dependent expressions for k(T) are provided in the two distinct T-ranges, excluding the upper limit obtained for k(177.5 K): In addition, the rate coefficients estimated by the canonical competitive unified statistical (CCUS) theory show a similar behaviour to the experimental results, when evaluated within the high-pressure limit. This is consistent with the experimentally observed independence of k(T) with total gas density at selected temperatures. Astrochemical networks, such as the KIDA database or UMIST, do not include the CHCN + OH reaction as a potential depletion process for acetonitrile in the ISM because the current studies predict very low rate coefficients at IS temperatures. According to the model (T = 10 K), the impact of the titled reaction on the abundances of CHCN appears to be negligible in dark molecular clouds of the ISM (∼1% of the total depletion reactions included in UMIST network). With respect to the potential formation of the CHCN radical in those environments, even

This study aims to assess the persistence of traditional journalistic quality features within the contemporary digital ecosystem. Digital advancements have fundamentally altered media creation, notably with the emergence of artificial intelligence (AI) in newsrooms. AI holds potential for substantial shifts in journalism, impacting business models, dissemination methods, and professional practices. Present in 75% of newsrooms, AI streamlines tasks, allowing more creative time for journalists. Ethical and quality concerns persist, particularly regarding AI’s ability to meet journalism’s established quality standards. This article aims to investigate the incorporation of these quality criteria in news articles generated by artificial intelligence (AI). Focus group and in-depth interviews were used as methodological techniques, involving ten experts. It is concluded that journalistic ethics have remained intact despite the disruptive technological advances in recent decades. However, there is a need to integrate these ethics with new criteria associated with the tools being used. Therefore, it is necessary to consider criteria from a dual perspective: both social and technological.

Cuenca, sus tierras, sus gentes, su pasado y su presente constituyen un sujeto de estudio fascinante al que, paradójicamente, no se le ha prestado la debida atención. Apenas ha concitado el interés de algunas reuniones científicas de mérito, que, así mismo, no han llegado a generar una continuidad suficiente para generar un espacio de debate dinámico, riguroso y en constante actualización, capaz de convocar la participación y la colaboración de investigadores consagrados y de jóvenes investigadores para comprender cada vez más y mejor ese pasado que hacemos presente. Este libro, y el congreso sobre el que descansan sus estudios, ha tenido por objeto contribuir a la generación de ese espacio desde una perspectiva rigurosamente científica, procurando profundizar en un pasado que corre desde la Prehistoria hasta la actualidad; siempre con la finalidad de servir de catalizador y difusor de un conjunto de propuestas científicas de altura que permitan aportar nuevas interpretaciones y un conocimiento más riguroso y profundo de este sujeto histórico que es Cuenca: sus tierras y sus gentes.

The funds of the Museum of Cuenca hold authentic collections of materials that come from archaeological despoilment and from donations and deposits made by private individuals. They constitute a valuable documentation of the cultural and patrimonial heritage of the province. Most of them remain unpublished. Among these materials, there is a group of metal objects, excluding coins, which are characterised by their typological heterogeneity. This volume presents a study of a total of 1.745 pieces, devoid of archaeological context. They have therefore been dated by formal parallels in the peninsular and foreign spheres, covering a chronological period between the Bronze Age and the Middle Ages., Los fondos del Museo de Cuenca custodian auténticas colecciones de materiales procedentes de la actuación de expoliadores clandestinos y de donaciones y entregas realizadas por particulares, que constituyen una valiosa documentación de la riqueza cultural y patrimonial de la provincia. La mayoría de ellos permanecen inéditos. Entre estos materiales, se encuentra un conjunto de objetos de metal, dejando fuera las monedas, que se caracterizan por su heterogeneidad tipológica. Presentamos en este volumen el estudio de un total de 1.745 piezas, desprovistas de contexto arqueológico. Por ello, han sido datadas por paralelos formales del ámbito peninsular y foráneo, que abarcan un período cronológico que se sitúa entre la Edad del Bronce y la Edad Media.

[EN] Classically, the immune response of the vertebrates has been divided into the innate response and the adaptative response, only the latter being able to build immunological memory. However, this dichotomic paradigm has been challenged by past and recent evidence that suggests that also the innate immune system can build nonspecific memory, which has been termed “trained immunity” (TRAIM). TRAIM can be defined as an enhanced state of the innate cells upon a first contact with a microorganism, a microbial component or a vaccine, which leads to an improved immune response in further encounters with related or unrelated pathogens. The Bacillus Calmette-Guérin (BCG) vaccine is a live attenuated Mycobacterium bovis (M. bovis) that was obtained through serial passages in culture medium until achieving an avirulent strain. For over a century, the BCG vaccine has been, and still is, the only licensed vaccine to prevent TB. Beyond its protective efficacy against this disease, numerous epidemiological surveys and experimental studies have attributed nonspecific effects to the BCG vaccine, inducing a cross-protection against unrelated infections through of a stimulation of the innate response. Currently, BCG has arisen as one of the most studied inducers of TRAIM, both in vitro and in vivo. However, live vaccines, such as BCG imply certain constraints, such as the necessity of maintaining the cold chain to ensure the viability of the live mycobacteria, and even certain risks for public health, such as adverse reactions upon parenteral administration, persistence of living microorganisms in the host tissues with possible excretion into the environment and interference with TB diagnostic tests. Conversely, heat-inactivated M. bovis preparations, that overcome the disadvantages of a live vaccine like BCG, have also demonstrated to protect against TB, partly through activation of the innate immunity. However, inactivated mycobacteria have received scarce attention in the frame, [ES] Clásicamente, la respuesta inmunitaria de los vertebrados diferenciaba entre respuesta innata y respuesta adaptativa, siendo solamente esta última capaz de desarrollar memoria inmunológica. Sin embargo, este paradigma dicotómico se ha visto cuestionado por evidencias pasadas y recientes que sugieren que también el sistema innato puede desarrollar memoria no específica, lo cual se ha denominado “inmunidad entrenada” (“trained immunity”, TRAIM). TRAIM se puede definir como un estado potenciado de las células innatas tras un primer contacto con un microorganismo, un componente microbiano o una vacuna, que conduce a una respuesta inmunitaria mejorada en posteriores encuentros con patógenos relacionados o no relacionados. La vacuna Bacilo de Calmette-Guérin (BCG) es una cepa viva atenuada de Mycobacterium bovis (M. bovis) que fue obtenida mediante pases seriados en medio de cultivo hasta lograr una cepa avirulenta. Durante más de un siglo, la vacuna BCG ha sido, y continúa siendo, la única vacuna autorizada para prevenir la tuberculosis (TB). Más allá de su eficacia protectora frente a la enfermedad, numerosos estudios epidemiológicos y experimentales atribuyen a la BCG efectos no específicos que inducen una protección cruzada frente a infecciones no relacionadas mediante la estimulación de la respuesta innata. Actualmente, la BCG se ha consolidado como uno de los inductores de TRAIM más estudiados, tanto in vitro como in vivo. Sin embargo, las vacunas vivas, como la BCG, implican ciertas limitaciones, como la necesidad de mantener la cadena de frío para asegurar la viabilidad de las micobacterias vivas, e incluso ciertos riesgos para la salud pública, tales como la persistencia de microorganismos vivos en los tejidos del hospedador con la posible excreción al medio o como la interferencia con las pruebas diagnósticas de TB. En cambio, las preparaciones de M. bovis inactivada por calor, que subsanan las desventajas de una vacuna viva como BCG, también han demostrado

[ES] La emergencia y re-emergencia de enfermedades arbovirales, su expansión geográfica y el aumento de la incidencia de estas en las últimas décadas constituye una de las principales preocupaciones en términos de salud pública y animal a escala global. Los virus del género Flavirirus transmitidos por mosquitos tienen, entre los arbovirus, una especial relevancia por su gran plasticidad genética, su diversidad, su demostrada capacidad de adaptación a diferentes escenarios y por ser la causa de numerosas enfermedades zoonóticas emergentes y re emergentes con alta incidencia e impacto clínico en personas y animales. El virus de West Nile es el modelo de Flavivirus expansivo, emergente y virulento que reúne la mayor parte de estas características, por lo que es fundamental comprender su eco-epidemiología, conocer la diversidad de vectores y hospedadores involucrados en su ciclo de vida en los diferentes territorios colonizados, así como las interacciones entre reservorios, patógeno, vectores y sus hospedadores que modulan su dinámica de transmisión. Esta Tesis Doctoral aborda la obtención de nuevos conocimientos sobre la ecología del virus West Nile en territorios continentales mediterráneos españoles de reciente colonización y escasamente estudiados. La hipótesis de trabajo general de la Tesis Doctoral es que el virus West Nile encuentra las condiciones favorables para persistir y prosperar en los escenarios en los que fauna silvestre, vectores y animales domésticos/personas coexisten en el interior continental de España. En la Introducción de la Tesis Doctoral se aborda exhaustivamente la taxonomía, diversidad, distribución y ecología de los Flavivirus, haciendo hincapié en el complejo de la encefalitis japonesa y las enfermedades que causan y, especialmente, en el virus de West Nile y los factores que modulan su dinámica de transmisión., [EN] The emergence and re-emergence of arboviral diseases, their geographical spread, and the increase in their incidence in recent decades constitute one of the main concerns in terms of public and animal health on a global scale. Among the arboviruses, mosquito-borne viruses of the Flavirirus genus are of particular relevance due to their high genetic plasticity, their diversity, their proven ability to adapt to different scenarios, and for being the cause of numerous emerging and re-emerging zoonotic diseases with high incidence and clinical impact in humans and animals. West Nile virus is the model of an expansive, emerging and virulent Flavivirus that combines most of these characteristics, so it is essential to understand its eco-epidemiology, to know the diversity of vectors and hosts involved in its life cycle in the different colonised territories, as well as the interactions between reservoirs, pathogen, vectors, and their hosts that modulate its transmission dynamics. This PhD Thesis aims to obtain new knowledge on the ecology of West Nile virus in recently colonised and poorly studied Spanish Mediterranean continental territories. The general working hypothesis of the PhD Thesis is that West Nile virus finds favourable conditions to persist and thrive in scenarios where wildlife, vectors and domestic animals/persons coexist in the Spanish continental interior. The Introduction of the PhD Thesis comprehensively addresses the taxonomy, diversity, distribution, and ecology of Flaviviruses, with emphasis on the Japanese encephalitis complex and the diseases it causes, and especially on West Nile virus and the factors that modulate its transmission dynamics.

Male infertility (MI) involves various endogenous and exogenous facts. These include oxidative stress (OS), which is known to alter several physiological pathways and it is estimated to be present at high levels in up to 80% of infertile men. That is why since the late 20th century, the relationship between OS and MI has been widely studied. New terms have emerged, such as Male Oxidative Stress Infertility (MOSI), which is proposed as a new category to define infertile men with high OS levels. Another important term is MOXI: Male, Antioxidants, and Infertility. This term refers to the hypothesis that antioxidants could improve male fertility without the use of assisted reproductive technology. However, there are no evidence-based antioxidant treatments that directly improve seminal parameters or birth ratio. In this regard, there is controversy about their use. While certain scientists argue against their use due to the lack of results, others support this use because of their safety profile and low price. Some uncertainties related to the use of antioxidants for treating MI are their questionable efficacy or the difficulties in knowing their correct dosage. In addition, the lack of quality methods for OS detection can lead to excessive antioxidant supplementation, resulting in "reductive stress". Another important problem is that, although the inflammatory process is interdependent and closely linked to OS, it is usually ignored. To solve these uncertainties, new trends have recently emerged. These include the use of molecules with anti-inflammatory and antioxidant potential, which are also able to specifically target the reproductive tissue; as well as the use of new methods that allow for reliable quantification of OS and a quality diagnosis. This review aims to elucidate the main uncertainties about MOXI and to outline the latest trends in research to develop effective therapies with clinically relevant outcomes.

Ticks are vectors of a great diversity of pathogens, mainly bacteria, protists, and viruses, among others. These pathogens are the causative agents of diseases in a wide range of vertebrates and have a great impact on the health of wild and domestic animals. Moreover, ticks and the animals they parasitize are a reservoir of these pathogens that can become a problem for humans, not only at the economic level but also in terms of public health, since many of them are considered zoonotic diseases in constant evolution and emergence.

Myxoma virus (MYXV) and rabbit hemorrhagic disease virus (RHDV) are important drivers of the population decline of the European rabbit, an endangered keystone species. Both viruses elicit strong immune responses, but the long-term dynamics of humoral immunity are imperfectly known. This study aimed to assess the determinants of the long-term dynamics of antibodies to each virus based on a longitudinal capture-mark-recapture of wild European rabbits and semiquantitative serological data of MYXV and RHDV GI.2-specific IgG. The study included 611 indirect enzyme-linked immunosorbent assay (iELISA) normalized absorbance ratios for each MYXV and RHDV GI.2 from 505 rabbits from 2018 to 2022. Normalized absorbance ratios were analyzed using log-linear mixed models, showing a significant positive relationship with the time since the first capture of individual rabbits, with monthly increases of 4.1% for antibodies against MYXV and 2.0% against RHDV GI.2. Individual serological histories showed fluctuations over time, suggesting that reinfections boosted the immune response and likely resulted in lifelong immunity. Normalized absorbance ratios significantly increased with the seroprevalence in the population, probably because of recent outbreaks, and with body weight, highlighting the role of MYXV and RHDV GI.2 in determining survival to adulthood. Juvenile rabbits seropositive for both viruses were found, and the dynamics of RHDV GI.2 normalized absorbance ratios suggest the presence of maternal immunity up to 2 months of age. Semiquantitative longitudinal serological data provide epidemiological information, otherwise lost when considering only qualitative data, and support a lifelong acquired humoral immunity to RHDV GI.2 and MYXV upon natural infection., [Importance] This study addresses the long-term dynamics of humoral immunity to two major viral pathogens of the European rabbit, an endangered keystone species of major ecological relevance. Such studies are particularly challenging in free-ranging species, and a combination of longitudinal capture-mark-recapture and semiquantitative serology was used to address this question. Over 600 normalized absorbance ratios of iELISA, obtained from 505 individual rabbits in 7 populations over 5 years, were analyzed using linear mixed models. The results support a lifelong acquired humoral immunity to myxoma virus and rabbit hemorrhagic disease virus upon natural infection and suggest the presence of maternal immunity to the latter in wild juvenile rabbits. These results contribute to understanding the epidemiology of two viral diseases threatening this keystone species and assist in developing conservation programs.

[Background]: Artificial insemination (AI) is a routine breeding technology in animal reproduction. Nevertheless, the temperature-sensitive nature and short fertile lifespan of ram sperm samples hamper its use in AI. In this sense, nanotechnology is an interesting tool to improve sperm protection due to the development of nanomaterials for AI, which could be used as delivery vehicles. In this work, we explored the feasibility of vitamin E nanoemulsion (NE) for improving sperm quality during transport., [Results]: With the aim of evaluating this proposal, ejaculates of 7 mature rams of Manchega breed were collected by artificial vagina and extended to 60 × 106 spz/mL in Andromed®. Samples containing control and NE (12 mmol/L) with and without exogenous oxidative stress (100 µmol/L Fe2+/ascorbate) were stored at 22 and 15 ºC and motility (CASA), viability (YO-PRO/PI), acrosomal integrity (PNA-FITC/PI), mitochondrial membrane potential (Mitotracker Deep Red 633), lipoperoxidation (C11 BODIPY 581/591), intracellular reactive oxygen species (ROS) production and DNA status (SCSA®) monitored during 96 h. Our results show that NE could be used to maintain ram spermatozoa during transport at 15 and 22 ºC for up to 96 h, with no appreciable loss of kinematic and physiological characteristics of freshly collected samples., [Conclusions]: The storage of ram spermatozoa in liquid form for 2-5 d with vitamin E nanoemulsions may lead more flexibility to breeders in AI programs. In view of the potential and high versatility of these nanodevices, further studies are being carried out to assess the proposed sperm preservation medium on fertility after artificial insemination.

[EN] Telomeres are repetitive DNA sequences at the end of chromosomes that protect them from degradation. They have been the focus of intense research because short telomeres would predict accelerated ageing and reduced longevity in vertebrates. Oxidative stress is considered a physiological driver of the telomere shortening and, consequently, short lifespan. Among molecules fighting against oxidative stress, glutathione is involved in many antioxidant pathways. Literature supports that oxidative stress may trigger a compensatory “hormetic” response increasing glutathione levels and telomere length. Here, we tested the link between total glutathione concentration and telomere length in captive birds (zebra finches; Taeniopygia guttata). Total glutathione levels were experimentally decreased during birds’ growth using a specific inhibitor of glutathione synthesis (buthionine sulfoximine; BSO). We monitored the birds’ reproductive performance in an outdoor aviary during the first month of life, and their longevity for almost 9 years. Among control individuals, erythrocyte glutathione levels during development positively predicted erythrocyte telomere length in adulthood. However, BSO-treated females, but not males, showed longer telomeres than control females in adulthood. This counterintuitive finding suggests that females mounted a compensatory response. Such compensation agrees with precedent findings in the same population where the BSO treatment increased growth and adult body mass in females but not males. BSO did not influence longevity or reproductive output in any sex. However, early glutathione levels and adult telomere length interactively predicted longevity only among control females. Those females with “naturally” low (non-manipulated) glutathione levels at the nestling age but capable of producing longer telomeres in adulthood seem to live longer. The results suggest that the capability to mount a hormetic response triggered by low early glutathione lev, [ES] Los telómeros son secuencias repetitivas de ADN que se encuentran al final de los cromosomas protegiéndolos de la degradación. Los telómeros han sido foco de intensa investigación ya que tener telómeros cortos predice un envejecimiento acelerado y una longevidad reducida en vertebrados. El estrés oxidativo se considera causante del acortamiento de los telómeros y, consecuentemente, una corta esperanza de vida. Entre las moléculas que luchan contra el estrés oxidativo, el glutatión está relacionado con numerosas vías metabólicas de otros antioxidantes. La literatura defiende que el estrés oxidativo podría desencadenar una respuesta hormética compensatoria, aumentando los niveles de glutatión y la longitud de los telómeros. En este trabajo testamos la relación entre los niveles de glutatión y la longitud de los telómeros en aves en condiciones de cautividad (diamantes mandarines; Taeniopygia guttata). Los niveles de glutatión fueron reducidos experimentalmente durante el desarrollo de las aves usando un inhibidor específico de la síntesis de glutatión (Buthionina sufolximina; BSO). El desempeño reproductivo de estas aves fue monitoreado en un aviario exterior durante los primeros meses de vida y la longevidad durante casi nueve años. Entre los individuos controles, los niveles de glutatión durante el desarrollo predijeron positivamente la longitud de los telómeros a edad adulta. Sin embargo, las hembras tratadas con BSO, pero no los machos, mostraron telómeros más largos a edad adulta que las hembras controles. Este hallazgo aparentemente contradictorio sugiere que las hembras desarrollaron una respuesta compensatoria. Tal compensación concuerda con los hallazgos anteriores obtenidos en la misma población: las hembras tratadas con BSO vieron aumentado su peso corporal a edad adulta, resultado no encontrado entre los machos. El tratamiento con BSO no afectó la longevidad o el desempeño reproductivo de ningún sexo. Sin embargo, los niveles de glutatión durante el d

The incidence of West Nile fever (WNF) is highly variable in emerging areas, making it difficult to identify risk periods. Using clinical case records has important biases in understanding the transmission dynamics of West Nile virus (WNV) because asymptomatic infections are frequent. However, estimating virus exposure in sentinel species could help achieve this goal at varying spatiotemporal scales. To identify the determinants of inter-annual variation in WNV transmission rates, we designed a 15-year longitudinal seroepidemiological study (2005–2020) in five environmentally diverse areas of southwestern Spain. We modeled individual annual area-dependent exposure risk based on potential environmental and host predictors using generalized linear mixed models. Further, we analyzed the weight of predictors on exposure probability by variance partitioning of the model components. The analysis of 2418 wild ungulate sera (1168 red deer - Cervus elaphus - and 1250 Eurasian wild boar - Sus scrofa) with a highly sensitive commercial blocking ELISA identified an average seroprevalence of 24.9% (95% confidence interval (CI): 23.2–26.7%). Antibody prevalence was slightly higher in wild boar (27.5%; CI: 25.1–30.1%) than in deer (22.2%; CI: 19.8–24.7%). We observed a spatial trend in exposure, with higher frequency in the southernmost areas and a slight, although area-dependent, increasing temporal trend. Host-related predictors were important drivers of exposure risk. The environmental predictor with the highest weight was annual cumulative precipitation, while temperature variations were also relevant but with less weight. We observed a coincidence of spatiotemporal changes in exposure with the notification of WNF outbreaks in horses and humans. That indicates the usefulness of wild ungulates as sentinels for WNV transmission and as models to understand its spatiotemporal dynamics. These results will allow the development of more accurate predictive models of spatiotemporal va

Ticks are the main vectors for the transmission of bacterial, protist and viral pathogens in Europe affecting wildlife and domestic animals. However, some of them are zoonotic and can cause serious, sometimes fatal, problems in human health. A systematic review in PubMed/MEDLINE database was conducted to determine the spatial distribution and host and tick species ranges of a selection of tick-borne bacteria (Anaplasma spp., Borrelia spp., Coxiella spp., and Rickettsia spp.), protists (Babesia spp. and Theileria spp.), and viruses (Orthonairovirus, and flaviviruses tick-borne encephalitis virus and louping ill virus) on the European continent in a five-year period (November 2017 – November 2022). Only studies using PCR methods were selected, retrieving a total of 429 articles. Overall, up to 85 species of the selected tick-borne pathogens were reported from 36 European countries, and Anaplasma spp. was described in 37% (159/429) of the articles, followed by Babesia spp. (34%, 148/429), Borrelia spp. (34%, 147/429), Rickettsia spp. (33%, 142/429), Theileria spp. (11%, 47/429), tick-borne flaviviruses (9%, 37/429), Orthonairovirus (7%, 28/429) and Coxiella spp. (5%, 20/429). Host and tick ranges included 97 and 50 species, respectively. The highest tick-borne pathogen diversity was detected in domestic animals, and 12 species were shared between humans, wildlife, and domestic hosts, highlighting the following zoonotic species: Anaplasma phagocytophilum, Babesia divergens, Babesia microti, Borrelia afzelii, Borrelia burgdorferi s.s., Borrelia garinii, Borrelia miyamotoi, Crimean-Congo hemorrhagic fever virus, Coxiella burnetii, Rickettsia monacensis and tick-borne encephalitis virus. These results contribute to the implementation of effective interventions for the surveillance and control of tick-borne diseases.

In the One Health context, Integrated Wildlife Monitoring (IWM) merges wildlife health monitoring (WHM) and host community monitoring to early detect emerging infections, record changes in disease dynamics, and assess the impact of interventions in complex multi-host and multi-pathogen networks. This study reports the deployment and results obtained from a nationwide IWM pilot test in eleven sites representing the habitat diversity of mainland Spain. In each study site, camera-trap networks and sampling of indicator species for antibody and biomarker analysis were used to generate information. The results allowed identifying differences in biodiversity and host community characteristics among the study sites, with a range of 8 to 19 relevant host species per point. The Eurasian wild boar (Sus scrofa) was the most connected and central species of the host communities, becoming a key target indicator species for IWM. A negative relationship between biodiversity and disease risk was detected, with a lower number and prevalence of circulating pathogens in the sites with more species in the community and larger network size. However, this overall trend was modified by specific host-community and environmental factors, such as the relative index of wild boar - red deer interactions or the proximity to urban habitats, suggesting that human-driven imbalances may favour pathogen circulation. The effort of incorporating wildlife population monitoring into the currently applied WHM programs to achieve effective IWM was also evaluated, allowing to identify population monitoring as the most time-consuming component, which should be improved in the future. This first nationwide application of IWM allowed to detect drivers and hotspots for disease transmission risk among wildlife, domestic animals, and humans, as well as identifying key target indicator species for monitoring. Moreover, anthropogenic effects such as artificially high wildlife densities and urbanisation were identi

There is a knowledge gap in the study of Argasidae soft ticks and the pathogens they can transmit. These hematophagous arthropods are widely distributed and are often considered typical bird ectoparasites. Tick-parasitized birds can act not only as a reservoir of pathogens but also can carry these pathogen-infected arthropods to new areas. Seven griffon vulture nestlings were sampled in northeastern Spain, collecting ticks (n = 28) from two individuals and blood from each vulture (n = 7). Blood samples from griffon vultures tested PCR positive for Flavivirus (7/7), Anaplasma (6/7), piroplasms (4/7), and Rickettsia (1/7). A total of 27 of the 28 analyzed ticks were positive for Rickettsia, 9/28 for Anaplasma, 2/28 for piroplasms, and 5/28 for Crimean-Congo Hemorrhagic fever virus (CCHFv). Sequencing and phylogenetic analyses confirmed the presence of Rickettsia spp., Babesia ardeae, and zoonotic Anaplasma phagocytophilum in vultures and Rickettsia spp., B. ardeae, and CCHFv genotype V in ticks.

[Objective]: Vector-borne diseases are a growing burden worldwide. In particular, the risks of allergic reactions to bites are associated with growing arthropod populations in contact with the public. The diversity of allergic reactions associated with host and arthropod factors difficult disease diagnosis, prognosis and prevention. Therefore, arthropod-associated allergies are underdiagnosed and require better surveillance of arthropod populations and disease diagnosis and management., [Methods]: To face these challenges, in this study, we describe five cases to illustrate arthropod-associated allergies with different symptomatology, including alpha-gal syndrome (AGS) associated with anti-alpha-gal IgE antibody titres. Information on symptoms in response to arthropod bites was collected from patients and medical doctors., [Results]: The five cases included patients bitten by a robber fly and different tick species. Cases were in Spain or U.S.A. Two cases were diagnosed with AGS and one case was diagnosed with anaphylaxis in response to tick bite with high anti-alpha-gal IgE levels. The symptoms in response to arthropod bites vary between different cases., [Conclusion]: Allergic reactions and symptoms in response to arthropod bites vary in association with host and arthropod factors. Herein we propose recommendations to control allergic symptoms, associated disease risk factors and the way forward to advance in the prevention and control of arthropod-associated allergies.

Tick microbiota can be targeted for the control of tick-borne diseases such as human granulocytic anaplasmosis (HGA) caused by model pathogen, Anaplasma phagocytophilum. Frankenbacteriosis is inspired by Frankenstein and defined here as paratransgenesis of tick symbiotic/commensal bacteria to mimic and compete with tick-borne pathogens. Interactions between A. phagocytophilum and symbiotic Sphingomonas identified by metaproteomics analysis in Ixodes scapularis midgut showed competition between both bacteria. Consequently, Sphingomonas was selected for frankenbacteriosis for the control of A. phagocytophilum infection and transmission. The results showed that Franken Sphingomonas producing A. phagocytophilum major surface protein 4 (MSP4) mimic pathogen and reduce infection in ticks by competition and interaction with cell receptor components of infection. Franken Sphingomonas-MSP4 transovarial and trans-stadial transmission suggests that tick larvae with genetically modified Franken Sphingomonas-MSP4 could be produced in the laboratory and released in the field to compete and replace the wildtype populations with associated reduction in pathogen infection/transmission and HGA disease risks.

We used primary care data to retrospectively describe the entry, spread, and impact of COVID-19 in a remote rural community and the associated risk factors and challenges faced by the healthcare team. Generalized linear models were fitted to assess the relationship between age, sex, period, risk group status, symptom duration, post-COVID illness, and disease severity. Social network and cluster analyses were also used. The first six cases, including travel events and a social event in town, contributed to early infection spread. About 351 positive cases were recorded and 6% of patients experienced two COVID-19 episodes in the 2.5-year study period. Five space-time case clusters were identified. One case, linked with the social event, was particularly central in its contact network. The duration of disease symptoms was driven by gender, age, and risk factors. The probability of suffering severe disease increased with symptom duration and decreased over time. About 27% and 23% of individuals presented with residual symptoms and post-COVID illness, respectively. The probability of developing a post-COVID illness increased with age and the duration of COVID-associated symptoms. Carefully registered primary care data may help optimize infection prevention and control efforts and upscale local healthcare capacities in vulnerable rural communities.

[Background]: Lyme borreliosis (LB) caused by Borrelia burgdorferi sensu lato complex spirochetes is one of the tick-borne diseases with high prevalence and social/economic burden in the United States, Spain, and other European countries. The objective is to address limited information available about the incidence, prevalence, and symptoms of LB, current prevention, and treatment interventions that are not adequately focused and thus not very effective against this disease., [Methods]: To address these limitations, in this study, we used a citizen science approach to evaluate the LB-associated risks and implementation of control interventions in Spain. A total of 405 participants in the survey were included in the analysis. Responses to the questionnaire were received during January-July 2022. The questionnaire contained qualitative and quantitative questions. Homogeneity among binary variables was analyzed using a Fisher's exact test., [Results]: Despite limitations of the study associated with response to the questionnaire and information on tick species, the results evidenced the effect of factors such as age, gender, tick bites, disease clinical signs, comorbidities such as alpha-gal syndrome, health care services, and treatment effectiveness affecting LB., [Conclusions]: The main conclusions of the study highlight the need for better surveillance of tick infestations, pathogen infection, and diagnosis of LB and related comorbidities. To advance in disease prevention, diagnosis, and treatment, new interventions need to be developed and implemented in both public and private health care services. Providing access to these results to the society, health care system, and scientists is important to further advance in disease surveillance, diagnosis, control, and prevention.

In the Anthropocene, many species are rapidly shifting their ranges in response to human-driven habitat modifications. Studying patterns and genetic signatures of range shifts helps to understand how species cope with environmental disturbances and predict future shifts in the face of global environmental change. We investigated the genetic signature of a contemporary wide-range expansion observed in the Iberian common vole Microtus arvalis asturianus shortly after a colonization event. We used mtDNA and microsatellite data to investigate patterns of genetic diversity, structure, demography, and gene flow across 57 localities covering the historical range of the species and the newly colonized area. The results showed a genetic footprint more compatible with a true range expansion (i.e. the colonization of previously unoccupied areas), than with a model of “colonization from within” (i.e. local expansions from small, unnoticed populations). Genetic diversity measures indicated that the source population was likely located at the NE of the historical range, with a declining gradient of genetic diversity towards the more recently invaded areas. At the expansion front, we observed the greatest gene flow and smallest pairwise differences between nearby localities. Both natural landscape features (rivers) and recent anthropogenic barriers (roads, railways) explained a large proportion of genetic variance among populations and had a significant impact on the colonization pathways used by voles.

Paratransgenesis through genetic manipulation of symbiotic or commensal microorganisms has been proposed as an effective and environmentally sound approach for the control of vector-borne diseases, including tick bite-related pathologies, and reducing pathogen transmission. Here, we present a protocol for Sphingomonas transformation with Anaplasma phagocytophilum major surface protein 4 and heat shock protein 70. We describe a step-by-step protocol for in vitro study of interactions between transformed Franken Sphingomonas and Ixodes scapularis ISE6 tick cells during A. phagocytophilum infection.

The Egyptian mongoose (Herpestes ichneumon) is considered a generalist and opportunistic predator whose European distribution is restricted to Iberia. However, all the previous studies on the Egyptian mongoose’s diet in the Iberian Peninsula have been performed in Portugal and Doñana National Park (southwestern Spain), covering a small region of the species’ current distribution. To improve the knowledge on the trophic ecology of the species, we estimated the feeding habits of the Egyptian mongoose in central Spain, its north-eastern range edge. Our study was based on the analysis of (1) scats (n = 251) and (2) stomach contents (n = 32); samples were collected between 2018 and 2022. We calculated the relative frequency of occurrence (%RFO) and the percentage consumed biomass (%CB) for 12 food groups across central Spain and we assessed site-seasonal variations in diet composition in three study areas. We also estimated rabbit densities in the three study areas to assess the effect of rabbit abundance on mongoose diets. Overall, our results showed that European rabbits were the group with highest %CB (30.7%) in the diet of the Egyptian mongoose, followed by reptiles (19.9%) and small mammals (19.9%). In the intensively studied areas, the %CB of rabbits was the highest in the area with the highest abundance of this prey, whereas reptiles had the highest %CB in the area with the lowest availability of rabbits. Diet diversity (Shannon index) varied between 1.73 and 2.47 in those areas and was inversely related to the availability of rabbits. Our results confirm the dietary flexibility of this predator that adapts its diet to the relative availability of different prey species in different habitats.

The area of Prados del Cervunal (PC) is an intra-morainic topographic depression located at 1800 m asl in the divide or interfluve between Garganta de Gredos and Garganta del Pinar valleys (Central Gredos; Iberian Central System, ICS). Both valleys, along with the adjacent Hoya Nevada, were occupied by glaciers during the Upper Pleistocene, leading to the development of the Prados del Cervunal moraine complexes studied in this work. Using cartographic methods and morphostratigraphic analysis, the three main glacial formations established in the Regional Chrono-Evolutionary Pattern for the ICS, Peripheral Deposits (PD), Principal Moraine (PM) and Internal Deposits (ID), have been identified and mapped in this area. The chronology of these formations has been implemented by Cosmic Ray Exposure (CRE) techniques using 10Be (new data) and 26Cl (previous data, recalculated in this work) in samples from morainic boulders. With these data, the following chrono-evolutionary sequence has been established: (stage 1) local-Maximum Ice Extent (MIE), dated in 25.0 ± 1.4 ka and corresponding to the maximum age obtained in these paleoglaciers; (stage 2) period of oscillations around the MIE, corresponding to the development of the PD Formation between ∼25 ka and ∼21 ka; (stage 3) period of readvance and stabilisation, dated after ∼21 ka (average age obtained for the PD moraines attached to PM moraines) and previous to ∼18 (minimum age obtained for a main crest of the PM formation); and (stage 4) onset of deglaciation dated around to ∼18 ka (average of ages obtained for the first main crest of the ID formation). During the stages of maximum ice expansion, these three glaciers formed an Ice field whose tongues were interconnected on the PC flat by an ice transfluence system (stages 1 and 2, Plateau Glacier Period). In later stages, the ice masses were partitioned, giving rise to valley glaciers and large moraines forming morainic complexes like those of PC (stages 2, 3 and 4, Valley

Parent–offspring conflict over food allocation can be modeled using two theoretical frameworks: passive (scramble competition) and active choice (signaling) resolution models. However, differentiating between these models empirically can be challenging. One possibility involves investigating details of decision-making by feeding parents. Different nestling traits, related to competitive prowess or signaling cryptic condition, may interact additively or non-additively as predictors of parental feeding responses. To explore this, we experimentally created even-sized, small broods of pied flycatchers and manipulated nestling cryptic quality, independently of size, by vitamin E supplementation. We explored how interactions between nestling cryptic condition, size, signals, and spatial location predicted food allocation and prey-testing by parents. Parents created the potential for spatial scramble competition between nestlings by feeding from and to a narrow range of nest locations. Heavier supplemented nestlings grew faster and were more likely to access profitable nest locations. However, the most profitable locations were not more contested, and nestling turnover did not vary in relation to spatial predictability or food supply. Postural begging was only predicted by nestling hunger and body mass, but parents did not favor heavier nestlings. This suggests that size-mediated and spatial competition in experimental broods was mild. Pied flycatcher fathers allocated food in response to nestling position and begging order, while mothers seemingly followed an active choice mechanism involving assessment of more complex traits, including postural intensity interacting with order, position, and treatment, and perhaps other stimuli when performing prey-testings. Differences in time constraints may underlie sex differences in food allocation rules.

West Nile virus (WNV) transmission rate is shaped by the interaction between virus reservoirs and vectors, which may be maximized in farm environments. Based on this hypothesis, we screened for WNV in wild birds in three scenarios with decreasing gradient of interaction with horses: (i) the farm (A1); (ii) the neighborhood (A2); and (iii) a wild area (A3). We captured wild birds and analyzed their sera for WNV antibodies by blocking ELISA and micro-virus neutralization test. Flavivirus infections were tested with generic and specific PCR protocols. We parameterized linear mixed models with predictors (bird abundance and diversity, vector abundance, vector host abundance, and weather quantities) to identify Flavivirus spp. and WNV exposure risk factors. We detected a low rate of Flavivirus infections by PCR (0.8%) and 6.9% of the birds were seropositive by ELISA. Exposure to Flavivirus spp. was higher in A1 (9%) than in A2 and A3 (5.6% and 5.8%, respectively). Bird diversity was the most relevant predictor of exposure risk and passerines dominated the on-farm bird community. Our results suggest that measures deterring the use of the farm by passerines should be implemented because the environmental favorability of continental Mediterranean environments for WNV is increasing and more outbreaks are expected.

Dual or multimodal imaging probes have emerged as powerful tools that improve detection sensitivity and accuracy in disease diagnosis by imaging techniques. Two imaging techniques that are complementary and do not use ionizing radiation are magnetic resonance imaging (MRI) and optical fluorescence imaging (OFI). Herein, we prepared metal-free organic species based on dendrimers with magnetic and fluorescent properties as proof-of-concept of bimodal probes for potential MRI and OFI applications. We used oligo(styryl)benzene (OSB) dendrimers core that are fluorescent on their own, and TEMPO organic radicals anchored on their surfaces, as the magnetic component. In this way, we synthesized six radical dendrimers and characterized them by FT-IR, 1H NMR, UV-Vis, MALDI-TOF, SEC, EPR, fluorimetry, and in vitro MRI. Importantly, it was demonstrated that the new dendrimers present two properties: on one hand, they are paramagnetic and show the ability to generate contrast by MRI in vitro, and, on the other hand, they also show fluoresce emission. This is a remarkable result since it is one of the very few cases of macromolecules with bimodal magnetic and fluorescent properties using organic radicals as the magnetic probe.

Knowledge of seasonal variation in hunting pressure and the demographic composition of hunting bags is likely to be important for the effective management of quarry species. Such knowledge is particularly important where regulatory mechanisms aim to avoid the over-exploitation of quarry species having unfavourable conservation status. We compiled information on the age composition of harvested European Turtle-doves (Streptopelia turtur), and the daily numbers of doves shot and retrieved from 68 hunting estates spread across four Spanish regions. We use these data to describe temporal variation, and identify factors affecting daily hunting bags, age composition and crippling losses. We found that more juvenile doves were hunted than adults (and more than expected in a stable population) especially at the beginning of the hunting season, suggesting greater vulnerability of juveniles to hunting. The number of doves hunted each day declined through the hunting season and increased with the number of hunters present. Crippling losses averaged 9.6 % of all doves shot; applying this rate to recent Spanish government hunting statistics suggests under-recording of approximately 90,000 doves annually for the entire country. Our data on age composition and crippling losses can serve as direct inputs into future models of sustainable harvest for Turtle Dove in Europe. Our results highlight the importance of improving knowledge about the variables influencing take at the hunting event level for designing more efficiency regulatory measures.

Overabundant populations of wild boar (Sus scrofa) are frequent where anthropogenic food is easily available and where hunting is limited. Improving recreational hunting efficacy may contribute to manage these populations. However, we do not know the factors modulating hunting efficacy in a hunting day (i.e., proportion of individuals culled from a population). Additionally, we need to explore which factors drive hunting efficacy in shooting opportunities (i.e., proportion of hunted individuals from a wild boar group in individual shooting opportunities). Here, we analyze the factors explaining wild boar hunting efficacy at the scale of driven hunt and shooting opportunity. Hunting efficacy was surveyed in 92 driven hunts (2017–2018 to 2020–2021 hunting seasons) and in 145 shooting opportunities in Sierra de Gredos (central Spain). We found that hunting efficacy at the scale of the driven hunt was mainly modulated by the number of hunting posts. Indeed, maximum levels of 82 posts showed a fourfold hunting efficacy (59%) compared to minimum levels reaching six hunting posts (16%). At the scale of the shooting opportunity, hunting efficacy was mainly driven by wild boar group size, being negatively related to hunting efficacy: the higher the number of individuals in the group, the lower the shooting efficacy (13-fold; 41% for an individual, 3% for individuals belonging to groups > 7 individuals). Given the need of maintaining native wild boar populations at densities compatible with biodiversity conservation, rural economy, and animal and public health, efforts towards facilitating more hunters per event would improve wild boar hunting efficacy.

European wild rabbit (Oryctolagus cuniculus) and red-legged partridge (Alectoris rufa) are main small game species of conservation concern in Mediterranean ecosystems. To date, their presence in wild boar’s (Sus scrofa) diet and factors driving their consumption have been little investigated. A genetic metabarcoding approach was used on 80 wild boar faeces collected from four hunting estates devoted to small game hunting during two different seasons. Abundances of wild boar, rabbits and partridges were first estimated. Results showed DNA of seventeen chordate species. The highest frequency of occurrence (FO) corresponded to mammals and birds, with 77.6 and 22.3%, respectively. DNA of game species was detected in 39/71 (FO = 55.0%) samples, highlighting the presence of European wild rabbit in 27 (FO = 38.0%) and red-legged partridge in eight (FO = 11.3%). Dietary composition varied between seasons and estates, being rabbit the main responsible (explaining 35.26% and 39.45% of differences, respectively). Rabbit FO in the diet was positively related to the abundance of wild boar and rabbit density on the estate. It was greater in autumn and in estates where rabbits were hunted. Regarding red-legged partridge, a significant and positive relationship between its population density and its diet FO was observed, without significant differences between seasons or estates. Overall, our results suggest wild boar as an opportunistic species whose diet is largely determined by the relative availability of different food resources. Its ecological role concerning small game species in Mediterranean agroecosystems seems to be more related to consumption of carrion during the hunting season than to direct predation.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) still poses a challenge for biomedicine and public health. To advance the development of effective diagnostic, prognostic, and preventive interventions, our study focused on high-throughput antibody binding epitope mapping of the SARS-CoV-2 spike RBD protein by IgA, IgM and IgG antibodies in saliva and sera of different cohorts from healthy uninfected individuals to SARS-CoV-2-infected unvaccinated and vaccinated asymptomatic, recovered, nonsevere, and severe patients. Identified candidate diagnostic (455-LFRKSNLKPFERD-467), prognostic (395-VYADSFVIRGDEV-407-C-KLH, 332-ITNLCPFGEV-342-C-KLH, 352-AWNRKRI-358-C-KLH, 524-VCGPKKSTNLVKN-536-KLH), and protective (MKLLE-487-NCYFPLQSYGFQPTNGVG-504-GGGGS-446-GGNYNYLYRLFRKSNLKPFERD-467) epitopes were validated with sera from prevaccine and postvaccine cohorts. The results identified neutralizing epitopes and support that antibody recognition of linear B-cell epitopes in RBD protein is associated with antibody isotype and disease symptomatology. The findings in asymptomatic individuals suggest a role for anti-RBD antibodies in the protective response against SARS-CoV-2. The possibility of translating results into diagnostic interventions for the early diagnosis of asymptomatic individuals and prognosis of disease severity provides new tools for COVID-19 surveillance and evaluation of risks in hospitalized patients. These results, together with other approaches, may contribute to the development of new vaccines for the control of COVID-19 and other coronavirus-related diseases using a quantum vaccinomics approach through the combination of protective epitopes.

The native Eurasian wild boar (Sus scrofa) is a relevant wildlife host for African swine fever (ASF) virus, contributing to infection maintenance and spread and representing a challenge for disease control. Combining published scientific evidence with expert opinion, we provide an updated global overview of ASF control in wild boar and feral pigs in different epidemiological scenarios. We synthesize current knowledge on key background aspects of wild boar ecology and management and on ASF epidemiology in wild boar and their relative, the feral pig. We propose that establishing a proper surveillance and monitoring scheme is a requisite for disease control in wildlife and that ASF and wild boar should be monitored in an integrated way, considering the changes in the host population as well as the spatial spread and temporal distribution of disease indicators, to make possible a critical assessment of the impact of interventions. The main body of the manuscript reviews the intervention options and ASF control attempts and their outcomes in different epidemiological situations from peacetime to endemicity. Current ASF control in wild boar relies on three essential tools: carcass destruction, wild boar culling, and fencing. The experience gained since the onset of the ongoing ASF pandemic shows that certain combinations of interventions can slow down ASF spread and eventually succeed in ASF eradication in wild boar, at least after point introductions. Several strengths and weaknesses of these strategies are identified.

Tuberculosis caused by Mycobacterium bovis and other related mycobacteria has been reported in a wide range of mammals worldwide. In the case of the Herpestidae family, Mycobacterium mungi and M. bovis, both belonging to the Mycobacterium tuberculosis Complex, have been reported in banded mongooses (Mungos mungo) in Africa and in Egyptian mongooses (Herpestes ichneumon) in Portugal, respectively. Thus, we hypothesized that Tuberculosis may occur in Egyptian mongooses from Spain. Twenty-five found dead Egyptian mongooses were necropsied in order to detect macroscopic TB-compatible lesions and mandibular lymph nodes and lungs were cultured onto mycobacteria-specific growth media. We isolated M. bovis in 3/25 Egyptian mongooses (12.00%, IC95: 4.17–29.96%) and identified spoligotypes SB0121 (2/3) and SB0134 (1). No macroscopic TB-compatible lesions were observed. To the best of our knowledge, this is the first report of M. bovis in Egyptian mongoose in Spain, as well as the only study that includes spolygotyping in this species. Although the absence of visible lesions suggests a minor role of the Egyptian mongoose in Tuberculosis epidemiology, further research thereon is encouraged.