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Abstract Sorting grids to exclude the juveniles of species targeted by bottom trawl fisheries from the catch are among the most promising solutions to reduce discards. We tested a two-sections Juveniles’ Sorting Grid (JSG) in a Mediterranean fisheries restricted area. First, we provided information on the vitality of individuals escaping from the JSG bars during towing, by analysing underwater footage. Then, we evaluated the catch performance of the JSG-equipped trawl compared to a standard trawl by analysing both the full species community in the catches and the main commercial species. The probability for individuals to be alive while escaping from JSG was always higher than 65% (on average), with some species (e.g. red mullet, gurnards, 91–99% on average) showing significantly higher probability than others (e.g. European hake, crustaceans, 65–82% on average). The installation of a JSG in the trawl net did not change the overall catch composition in the codend, although significant differences were observed at the single species level. The JSG was effective at reducing undersized individuals of European hake, although a loss of legal-sized individuals was observed due to escapement. A significantly lower retention of the JSG-equipped trawl was also observed for other commercial species, such as deep-water rose shrimp and broadtail shortfin squid.

This study presents an integrated approach that combines the Levenberg–Marquardt (LM) and Luus–Jaakola (LJ) algorithms to enhance parameter estimation for various applications. The LM algorithm, known for its precision in solving non-linear least squares problems, is effectively paired with the LJ algorithm, a robust stochastic optimization method, to improve accuracy and computational efficiency. This hybrid LM-LJ methodology is demonstrated through several case studies, including the optimization of MESH equations in distillation processes, modeling controlled diffusion in biopolymer films, and analyzing heat and mass transfer during the drying of cylindrical quince slices. By overcoming the convergence issues typical of gradient-based methods and performing global searches without initial parameter bounds, this approach effectively handles complex models and closely aligns simulation results with experimental data. These capabilities highlight the versatility of this approach in engineering and environmental modeling, significantly enhancing parameter estimation in systems governed by differential equations.

Amoebiasis is a parasitic infection that represents a public health problem in developing countries including Asia and Latin America where it is endemic (1000–5000 cases/100,000 habitants/year). The majority of patients have an asymptomatic course; however, 10% of patients develop complications with high morbidity and mortality, such as colonic perforation or fulminant amoebic colitis. We report a case in which a 73-year-old female presented with an acute abdomen that was initially attributed to a bowel obstruction that rapidly progressed to fulminant colitis with bowel perforation requiring total colectomy. Pre-surgical endoscopic histopathological examination revealed findings suggestive of Entamoeba histolytica trophozoites that were later confirmed in the colon post-surgical specimen leading to a diagnosis of fulminant amoebic colitis. This atypical presentation of amoebiasis, further expands the already broad differential diagnosis of acute abdominal pathology in the elderly population. A high index of suspicion is required for its prompt treatment and to prevent life-threatening complications.

Long-term and culture-specific experience of music shapes rhythm perception, leading to enculturated expectations that make certain rhythms easier to track and more conducive to synchronized movement. However, the influence of enculturated bias on the moment-to-moment dynamics of rhythm tracking is not well understood. Recent modeling work has formulated entrainment to rhythms as a formal inference problem, where phase is continuously estimated based on precise event times and their correspondence to timing expectations: PIPPET (Phase Inference from Point Process Event Timing). Here we propose that the problem of optimally tracking a rhythm also requires an ongoing process of inferring which pattern of event timing expectations is most suitable to predict a stimulus rhythm. We formalize this insight as an extension of PIPPET called pPIPPET (PIPPET with pattern inference). The variational solution to this problem introduces terms representing the likelihood that a stimulus is based on a particular member of a set of event timing patterns, which we initialize according to culturally-learned prior expectations of a listener. We evaluate pPIPPET in three experiments. First, we demonstrate that pPIPPET can qualitatively reproduce enculturated bias observed in human tapping data for simple two-interval rhythms. Second, we simulate categorization of a continuous three-interval rhythm space by Western-trained musicians through derivation of a comprehensive set of priors for pPIPPET from metrical patterns in a sample of Western rhythms. Third, we simulate iterated reproduction of three-interval rhythms, and show that models configured with notated rhythms from different cultures exhibit both universal and enculturated biases as observed experimentally in listeners from those cultures. These results suggest the influence of enculturated timing expectations on human perceptual and motor entrainment can be understood as approximating optimal inference about the rhythmic stimulus, with respect to prototypical patterns in an empirical sample of rhythms that represent the music-cultural environment of the listener.

Presentamos un resumen de la investigación que estamos llevando a cabo a través del proyecto internacional “They:Live - Student lives revealed through context-based art practices” en el marco del programa de ayudas de Europa Creativa y en el que participan seis países europeos. El objetivo fue recopilar fotografías de la vida de los estudiantes en diversos campus universitarios desde la II Guerra Mundial hasta la actualidad. De entre todo el corpus recopilado hicimos una selección a partir de las cuales realizamos entrevistas a sus protagonistas para que nos relatasen la intrahistoria de las fotos seleccionadas (relato oral). Todas estas fotos y entrevistas se documentaron en el repositorio de acceso abierto en línea Topoteka. A continuación, lanzamos una convocatoria competitiva para organizar unas Residencias Artísticas (AiR, Artist in Residence) en la que en cada país participante eligió un par creativo de artista y comisario que tuvo que proponer un proyecto artístico contextual que tuviese en cuenta estas fotos y entrevistas —y que se está llevando a cabo en los campus universitarios participantes (febrero-mayo de 2002). Previamente, estos pares creativos realizaron una estancia formativa en el Institut für Kunst im Kontex, Universität der Künste Berlin para recibir formación en arte contextual. A continuación, se realizarán exposiciones itinerantes (mediados de 2022). Se trata, pues, de un método interdisciplinar con aplicación de prácticas artísticas contextuales. Todo ello con el objetivo de subrayar cómo la vida de los estudiantes y sus actividades representan un reservorio vivo de ideas y relaciones innovadoras, una fuente a través de la cual se puede seguir un desarrollo evolutivo del patrimonio intelectual y puede establecerse una visión completamente nueva de la cultura europea y su desarrollo futuro. Los resultados de nuestro proyecto servirán como directrices para gerentes de instituciones culturales y galerías de arte, comisarios y artistas para la mejora de enfoques innovadores para el desarrollo de audiencias jóvenes. Todo ello cristalizará en un libro editado en inglés en formato Open Access (OA) que publicará la editorial académica británica Intellect.

Grasp stability prediction of unknown objects is crucial to enable autonomous robotic manipulation in an unstructured environment. Even if prior information about the object is available, real-time local exploration might be necessary to mitigate object modelling inaccuracies. This paper presents an approach to predict safe grasps of unknown objects using depth vision and a dexterous robot hand equipped with tactile feedback. Our approach does not assume any prior knowledge about the objects. First, an object pose estimation is obtained from RGB-D sensing; then, the object is explored haptically to maximise a given grasp metric. We compare two probabilistic methods (i.e. standard and unscented Bayesian Optimisation) against random exploration (i.e. uniform grid search). Our experimental results demonstrate that these probabilistic methods can provide confident predictions after a limited number of exploratory observations, and that unscented Bayesian Optimisation can find safer grasps, taking into account the uncertainty in robot sensing and grasp execution.

Site selection of health facilities is critical in ensuring universal access to basic healthcare services. However, in many low and middle-income countries (LMICs) like the Philippines, site selection is traditionally based on political and pragmatic considerations. Moreover, literature that demonstrates the application of facility location models in the Philippine healthcare setting remains scarce, and their usage in actual facility planning is even more limited. In this study, we proposed a variation of cooperative covering maximal models to identify the optimal location of primary care facilities. We demonstrated the feasibility of implementing such a model by using open source data on an actual city in the Philippines. Our results generated multiple candidate locations of primary care facilities depending on the equity and efficiency parameters. This approach could be used as one of the critical considerations in evidence-based, multi-criterion health facility location decisions of governments, and can also be adapted in other industries, given the model's use of readily available open source datasets.

Plants are movers, but the nature of their movement differs dramatically from that of creatures that move their whole body from point A to point B. Plants grow to where they are going. Bio-inspired robotics sometimes emulates plants' growth-based movement; but growing is part of a broader system of movement guidance and control. We argue that ecological psychology's conception of “information” and “control” can simultaneously make sense of what it means for a plant to navigate its environment and provide a control scheme for the design of ecological plant-inspired robotics. In this effort, we will outline several control laws and give special consideration to the class of control laws identified by tau theory, such as time to contact.

Dealing safely with nuclear waste is an imperative for the nuclear industry. Increasingly, robots are being developed to carry out complex tasks such as perceiving, grasping, cutting, and manipulating waste. Radioactive material can be sorted, and either stored safely or disposed of appropriately, entirely through the actions of remotely controlled robots. Radiological characterisation is also critical during the decommissioning of nuclear facilities. It involves the detection and labelling of radiation levels, waste materials, and contaminants, as well as determining other related parameters (e.g., thermal and chemical), with the data visualised as 3D scene models. This paper overviews work by researchers at the QMUL Centre for Advanced Robotics (ARQ), a partner in the UK EPSRC National Centre for Nuclear Robotics (NCNR), a consortium working on the development of radiation-hardened robots fit to handle nuclear waste. Three areas of nuclear-related research are covered here: human–robot interfaces for remote operations, sensor delivery, and intelligent robotic manipulation.

Tactile sensing is crucial for robots to manipulate objects successfully. However, integrating tactile sensors into robotic hands is still challenging, mainly due to the need to cover small multi-curved surfaces with several components that must be miniaturized. In this paper, we report the design of a novel magnetic-based tactile sensor to be integrated into the robotic hand of the humanoid robot Vizzy. We designed and fabricated a flexible 4 × 2 matrix of Si chips of magnetoresistive spin valve sensors that, coupled with a single small magnet, can measure contact forces from 0.1 to 5 N on multiple locations over the surface of a robotic fingertip; this design is innovative with respect to previous works in the literature, and it is made possible by careful engineering and miniaturization of the custom-made electronic components that we employ. In addition, we characterize the behavior of the sensor through a COMSOL simulation, which can be used to generate optimized designs for sensors with different geometries.

Humanoid robots are resourceful platforms and can be used in diverse application scenarios. However, their high number of degrees of freedom (i.e., moving arms, head and eyes) deteriorates the precision of eye-hand coordination. A good kinematic calibration is often difficult to achieve, due to several factors, e.g., unmodeled deformations of the structure or backlash in the actuators. This is particularly challenging for very complex robots such as the iCub humanoid robot, which has 12 degrees of freedom and cable-driven actuation in the serial chain from the eyes to the hand. The exploitation of real-time robot sensing is of paramount importance to increase the accuracy of the coordination, for example, to realize precise grasping and manipulation tasks. In this code paper, we propose an online and markerless solution to the eye-hand kinematic calibration of the iCub humanoid robot. We have implemented a sequential Monte Carlo algorithm estimating kinematic calibration parameters (joint offsets) which improve the eye-hand coordination based on the proprioception and vision sensing of the robot. We have shown the usefulness of the developed code and its accuracy on simulation and real-world scenarios. The code is written in C++ and CUDA, where we exploit the GPU to increase the speed of the method. The code is made available online along with a Dataset for testing purposes.

The first World Cup in history took place in July 1930 in Uruguay. The country, which had won a few years back the football gold at Colombes (1924) and Amsterdam (1928) Olympic Games, already enjoyed a solid reputation in this sport. These international successes had triggered a massive passion in the country for the game. The holding of the World Cup in Montevideo, supported by the government, met a strong enthusiasm among the population with the exception of certain political sectors. Among those, Uruguay's Communist Party chose to totally reject the government's project while at the same time promoting an alternative model for sports, directly inspired by the Soviet Union's example. With the analysis of this communist stance we seek to participate in the efforts to historicize the first edition of the now FIFA's flagship tournament. This important sporting event, scarcely studied by current historiography, is yet fundamental given its leading role in the establishment, during the interwar period, of world's biggests sports competitions.

El presente trabajo de investigación se ha planteado en el marco de una beca para alumnos avanzados de la UNPA, Unidad Académica Río Gallegos. Tiene como objetivo analizar las variaciones temporales de la población del mejillón Mytilus edulis platensis y de la almeja Darina solenoides y se inserta en un Proyecto de Investigación (29A/227 dirigido por Dra. Z. Lizarralde) que tiene como objeto estudiar la ecología trófica del Ostrero Austral (Haematopus leucopodus) en el estuario del río Gallegos (Santa Cruz). Durante el 2010 se efectuaron muestreos mensuales (abril a diciembre) en un banco de mejillones y otro de almejas, con el objeto de analizar la variación temporal de la densidad y biomasa, y la estructura de tallas poblacional. Se analizó la granulometría y porcentaje de materia orgánica de los sedimentos. El sedimento del banco de mejillones se encuentra constituido en su mayoría por la fracción correspondiente a los gránulos; en el banco de almejas por arenas muy finas (43%) y finas (39%). En ambos casos la materia orgánica no supera el 3 %. La densidad de mejillones alcanzó un valor máximo de 2255 individuos/m2 en abril y un mínimo de 497 en noviembre. La biomasa máxima se encontró en abril (110 g/m 2). La densidad de Darina solenoides alcanzó un valor máximo de 565 en junio y un mínimo de 282 individuos/m2 en el mes de octubre. La biomasa máxima ocurrió en diciembre (132 g/m2). No se detectó el período de reclutamiento de nuevos individuos en los bancos de las especies estudiadas.

The concept of affordance perception is one of the distinctive traits of human cognition; and its application to robots can dramatically improve the quality of human-robot interaction (HRI). In this paper we explore and discuss the idea of “emotional affordances„ by proposing a viable model for implementation into HRI; which considers allocentric and multimodal perception. We consider “2-ways„ affordances: perceived object triggering an emotion; and perceived human emotion expression triggering an action. In order to make the implementation generic; the proposed model includes a library that can be customised depending on the specific robot and application scenario. We present the AAA (Affordance-Appraisal-Arousal) model; which incorporates Plutchik’s Wheel of Emotions; and we outline some numerical examples of how it can be used in different scenarios.

Mediante el análisis textual de la coda fotográfica en b/n del film Vozvrashcheniye (El regreso, Andréi Zvyagintsev, 2003) proponemos una hipótesis sobre el secreto que guarda el cofre que el padre, ausente de la familia tras doce años, busca en un viaje iniciático en compañía de sus hijos y lejos de la madre. Ello nos llevará a evaluar el papel simbólico del padre en relación a la madre y a la dolorosa y necesaria adquisición de libertad por parte de sus hijos. Matizaremos los análisis y lecturas predominantes al respecto, principalmente de cariz feminista y político, que se han hecho de este film post-soviético. Para ello, entrecruzaremos la Teoría del Texto preconizada por el profesor Jesús González Requena y su grupo de investigación, Trama y Fondo, del que el autor forma parte. En este sentido, el propio director nunca se ha pronunciado por el verdadero contenido del cofre; pero pensamos que tiene una buena razón para ello. Palabras clave: Vozvrashcheniye; Zvyagintsev; mito; análisis textual; fotografía. Keywords: Vozvrashcheniye; Zvyagintsev; Myth; Textual Analysis, Photography.

This paper presents an easy means to produce a 3-axis Hall effect–based skin sensor for robotic applications. It uses an off-the-shelf chip and is physically small and provides digital output. Furthermore, the sensor has a soft exterior for safe interactions with the environment; in particular it uses soft silicone with about an 8 mm thickness. Tests were performed to evaluate the drift due to temperature changes, and a compensation using the integral temperature sensor was implemented. Furthermore, the hysteresis and the crosstalk between the 3-axis measurements were evaluated. The sensor is able to detect minimal forces of about 1 gf. The sensor was calibrated and results with total forces up to 1450 gf in the normal and tangential directions of the sensor are presented. The test revealed that the sensor is able to measure the different components of the force vector.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infects various mammalian species, with farmed minks experiencing the highest number of outbreaks. In Spain, we analyzed 67 whole genome sequences and eight spike sequences from 18 outbreaks, identifying four distinct lineages: B.1, B.1.177, B.1.1.7, and AY.98.1. The potential risk of transmission to humans raises crucial questions about mutation accumulation and its impact on viral fitness. Sequencing revealed numerous not-lineage-defining mutations, suggesting a cumulative mutation process during the outbreaks. We observed that the outbreaks were predominantly associated with different groups of mutations rather than specific lineages. This clustering pattern by the outbreaks could be attributed to the rapid accumulation of mutations, particularly in the ORF1a polyprotein and in the spike protein. Notably, the mutations G37E in NSP9, a potential host marker, and S486L in NSP13 were detected. Spike protein mutations may enhance SARS-CoV-2 adaptability by influencing trimer stability and binding to mink receptors. These findings provide valuable insights into mink coronavirus genetics, highlighting both host markers and viral transmission dynamics within communities.

Background: The European bison (Bison bonasus), a symbol of Polish nature, is a protected species that requires active health monitoring. However, conservation efforts are made difficult by the zoonotic diseases such as brucellosis and tuberculosis., Objective: The aim of this study was to screen the Polish European bison population for exposure to the Mycobacterium tuberculosis complex (MTC) and Brucella spp., Methods: A total of 323 free-living and captive European bison from 13 localities were tested serologically for antibodies against the M. bovis P22 multi-protein complex (in-house ELISA) and against Brucella spp. (commercial ELISA)., Results: Antibodies against the MTC (P22) were detected in 7% (22/323) of the tested European bison. Anti-MTC antibody positivity was not significantly different by sex, age, and captive/free range status. Anti-MTC antibodies were found in six of 13 populations sampled, always in populations with larger sample sizes including the four free-living ones. Antibodies against Brucella spp. were detected in 36% (116/323) of the tested bison. While Brucella spp. antibody prevalence was not different by sex, it was significantly different by age (lower in adults) and captive/free-living status. Brucella spp. seroprevalence decreased with sample size and seropositive bison were found in 12 of 13 sampling populations., Conclusions: Our findings identify potential emerging threats to the European bison population and confirm the first serological response to P22 in European bison. As Poland is currently officially free of brucellosis and bovine tuberculosis, our results require careful interpretation. Further studies are needed to establish the presence of cross-reactions with atypical mycobacteria in the case of MTC and other bacteria (e.g. Yersinia enterocolitica O:9) in the case of Brucella spp., (© 2023 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

The identification of vertebral fracture is a key point in an FLS. We have analyzed the characteristics of 570 patients according to the route of identification (referral by other doctors, emergency registry or through VFA), concluding that promoting referral by other doctors with a training campaign is effective., Purpose: Vertebral fractures (VF) are associated with increased risk of further VFs. Our objective was to analyze the characteristics of patients with VF seen in a Fracture Liaison Service (FLS)., Methods: An observational study was carried out on patients with VF referred to the outpatient metabolic clinic (OMC) after a training campaign, identified in the emergency registry, and captured by VF assessment with bone densitometry (DXA-VFA) in patients with non-VFs. Patients with traumatic VF or VF > 1 year, infiltrative or neoplastic disease were excluded. The number and severity of VFs (Genant) were analyzed. Treatment initiation in the first 6 months after baseline visit was reviewed., Results: Overall, 570 patients were included, mean age 73. The most common route for identifying VF was through referral to OMC (303 cases), followed by the emergency registry (198) and DXA-VFA (69). Osteoporosis by DXA was found in 312 (58%) patients and 259 (45%) had ≥ 2 VFs. The rate of grade 3 VFs was highest among patients on the emergency registry. Those identified through OMC had a higher number of VFs, a higher rate of osteoporosis, more risk factors and greater treatment initiation. Patients with VFs detected by DXA-VFA were mostly women with a single VF and had a lower rate of osteoporosis by DXA., Conclusions: We present the distribution of VFs by the route of identification in an FLS. Promoting referral by other doctors with a training campaign may help in the quality improvement of the FLS-based model of care., (© 2023. The Author(s).)

Tuberculosis (TB) is a multi-host infectious disease caused by members of the Mycobacterium tuberculosis complex (MTC). In Mediterranean ecosystems, where multiple animal hosts of TB are present, identifying the role of the different species involved in the epidemiology of TB is a key point to be able to implement proper control measures. Sheep are susceptible to MTC infection but have traditionally been considered a spillover host. However, the occurrence of outbreaks involving sheep in recent years evidences the need to better understand the role of this small ruminant species in the epidemiology of the disease. Here, we aimed to determine the seroprevalence and risk factors associated with MTC seropositivity in sheep in Andalusia (southern Spain), a region with one of the highest prevalence of MTC infection in both cattle and wild ungulates. A total of 2266 sheep from 83 flocks were tested for antibodies against MTC using an in-house indirect ELISA. Anti-MTC antibodies were detected in 16 (0.7%) of the 2266 sheep (adjusted true prevalence 0.29%, 95% posterior probability interval 0.01-1.05). Seropositivity was found in 14.5% (12/83; 95%CI: 6.9-22.0) of the sheep farms analyzed. A semi-extensive management system was identified as a risk factor associated with MTC seropositivity in sheep farms (OR = 3.7; p < 0.038; 95%CI: 1.1-12.4) in the study area. To the best of the authors' knowledge, this is the first active TB surveillance study carried out to assess MTC exposure in sheep. Our results indicate MTC circulation in sheep farms in southern Spain. However, the low individual seroprevalence obtained suggests that sheep may play a limited role in the epidemiology of TB in this region. Serosurveillance programs could be a valuable tool to detect MTC circulation in sheep in risk scenarios or target farms, in order to optimize control measures on TB animal in multi-host Mediterranean ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Previous studies have shown that dopaminergic activity modulates the salience of novel stimuli enabling the formation of recognition memories. In this work, we hypothesize that dopamine released into the insular cortex (IC) from the ventral tegmental area (VTA) inputs enables the acquisition to consolidate object recognition memory. It has been reported that short training produces weak recognition memories; on the contrary, longer training produces lasting and robust recognition memories. Using a Cre-recombinase under the tyrosine hydroxylase (TH+) promoter mouse model, we photostimulated the VTA-IC dopaminergic pathway during short training or photoinhibited the same pathway during long training while mice explored objects. Our results showed that the photostimulation of the VTA-IC pathway during a short training enables the acquisition of recognition memory. Conversely, photoinhibition of the same pathway during a long training prevents the acquisition of recognition memory. Interestingly, the exploration time of the objects under photoinhibition or photostimulation of the dopaminergic VTA-IC pathway was not altered. Significantly, this enhancement of acquisition of the object recognition memory through the photostimulation of the VTA dopaminergic neurons could be impaired by the blockage of the D1-like receptors into the IC, either before or after the photostimulation. Altogether, our results suggest that dopamine released by the VTA is required during the acquisition to consolidate the object recognition memory through D1-like receptors into the IC without affecting the activity or the motivation to explore objects., Competing Interests: Declaration of competing interest None., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The P22 ELISA was recently developed for the serodiagnosis of animal tuberculosis. Herein, the stability of the P22 antigen in different presentations and storage conditions, and the cross-reactivity with Corynebacterium pseudotuberculosis infection in small ruminants were evaluated. For the stability assay, serum samples from cows, sheep, goats, alpacas, badgers, and wild boar were used in the P22 ELISA. The cross-reactivity analysis used sera from sheep and goats with caseous lymphadenitis (CLA). Differences in the immune recognition of P22 were found when the antigen was stored at 40 °C, but without altering the negative or positive status of each sample. P22 ELISA presented 5.71 % cross-reactivity when CLA-positive sheep were evaluated, but no cross-reaction was observed among CLA-positive goat serum samples. This study showed that the P22 protein complex is stable under different formulations and temperatures, and that the assay presents a low cross-reactivity with CLA., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Caprine tuberculosis (TB) is a zoonosis caused by members of the Mycobacterium tuberculosis complex (MTBC). Caprine TB eradication programmes are based mainly on intradermal tuberculin tests and slaughterhouse surveillance. However, the use of serological test has been extended as a potential diagnostic tool in goats through the use of serum, plasma, or even milk samples. Milk production and the antibodies (Ab) present in milk can vary depending on several circumstances. In the present study, different factors that may affect the performance of humoral TB diagnosis were analysed using goat milk samples: 1) lactation stage, 2) a recent previous skin test (booster effect) and 3) the effect of freeze-thaw cycles on milk samples preserved with azidiol. TB-infected animals (n = 44) were selected to evaluate the evolution of the Ab levels during the 6-month lactation period, along with its potential effect on the P22 ELISA results. In general, no significant changes (p = 0.079) were observed throughout the study as regards Ab levels in milk samples between consecutive analysis although the reactivity to P22 ELISA decreased when samplings were performed at the last two months of the lactation. Regarding the booster effect, the quantitative results showed a significant variation (p < 0.001) for both milk and serum samples when serological tests were carried out 15 days after the skin test. Finally, there were no significant differences (p = 0.99) in the P22 ELISA results when using milk samples preserved with azidiol that had undergone freeze-thaw cycles., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Caprine tuberculosis (TB) is a zoonosis caused by members of the Mycobacterium tuberculosis complex (MTBC). Caprine TB control and eradication programmes have traditionally been based on intradermal tuberculin tests and slaughterhouse surveillance. However, this strategy has limitations in terms of sensitivity and specificity. Different factors may affect the performance of the TB diagnostic tests used in goats and, subsequently, the detection of TB-infected animals. In the present study, the effect of two of the factors that may affect the performance of the techniques used to diagnose TB in goats, the topical administration of corticosteroids and a recent pre-sensitisation with tuberculin, was analysed., Methods: The animals (n = 151) were distributed into three groups: (1) a group topically treated with corticosteroids 48 h after intradermal tuberculin tests (n = 53); (2) a group pre-sensitised with bovine and avian purified protein derivatives (PPDs) 3 days before the intradermal tuberculin test used for TB diagnosis (n = 48); and (3) a control group (n = 50). All the animals were tested using single and comparative intradermal tuberculin (SIT and CIT, respectively) tests, an interferon-gamma release assay (IGRA) and a P22 ELISA., Results: The number of SIT test reactors was significantly lower in the group treated with corticosteroids when compared to the pre-sensitised (p < 0.001) and control (p = 0.036) groups. In contrast, pre-sensitisation with bovine and avian PPDs did not cause a significant reduction in the number of SIT and CIT test reactors compared with the control group. In fact, a higher number of reactors was observed after the prior tuberculin injection in the pre-sensitised group (p > 0.05). No significant effect was observed on IGRA and P22 ELISA due to corticosteroids administration. Nevertheless, a previous PPD injection affected the IGRA performance in some groups., Conclusions: The application of topical corticosteroid 24 h before reading the SIT and CIT tests can reduce the increase in skin fold thickness and subsequently significantly decrease the number of positive reactors. Corticosteroids used can be detected in hair samples. A previous pre-sensitisation with bovine and avian PPDs does not lead to a significant reduction in the number of intradermal tests reactors. These results are valuable in order to improve diagnosis of caprine TB and detect fraudulent activities in the context of eradication programs., (© 2022. The Author(s).)

The single and comparative intradermal tuberculin (SIT and CIT) tests are used for the ante-mortem diagnosis of caprine tuberculosis (TB). The tuberculin injection site has been associated with a different performance of the test in cattle. In contrast to that required in cattle in Europe (cervical injection), it can be carried out in the scapular region in goats. Nevertheless, there are no previous data concerning the effect of the injection site on the performance of the test in goats. The aim of the present study was to evaluate the effect of two different inoculation sites (cervical and scapular) on the performance of the SIT/CIT tests. This was done by intradermally inoculating 309 goats from two infected herds and one TB-free herd with both avian and bovine PPDs in the mid-cervical and scapular regions. None of the animals from the TB-free herd had positive reactions, and the number of reactors was not significantly higher, regardless of the inoculation site, in the high and low prevalence herds. However, significantly higher increases in skin fold thickness were observed on the cervical site when compared to the scapular site after the avian and bovine PPD inoculations in the TB-free herd ( p < 0.001) and after the bovine PPD injection in the high prevalence herd ( p = 0.003). The presence of clinical signs was also more evident on the cervical site when using avian and bovine PPDs in the high prevalence herd ( p < 0.01). In contrast, increases in higher skin fold thickness were observed on the scapular site when compared to the cervical site after the bovine and avian PPD inoculations were employed in the low prevalence herd ( p < 0.01). These results suggest that the cervical injection of PPDs may improve the sensitivity of the intradermal tuberculin test in high TB prevalence caprine herds, mainly owing to the increased presence of local clinical signs and a better performance of the CIT test. Moreover, specificity was not affected when using standard interpretations, although further analyses in a great number of herds are required in order to confirm these findings., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ortega, Roy, Álvarez, Sánchez-Cesteros, Romero, Infantes-Lorenzo, Sáez, López, Domínguez, de Juan and Bezos.)

The ante-mortem diagnosis of tuberculosis (TB) in ruminants is based mainly on the intradermal tuberculin test and the IFN-γ assay. Antibody (Ab)-based tests have emerged as potential tools for the detection of TB infected animals using serum, plasma, or even milk samples. Oral fluids have also been evaluated as alternative samples with which to detect specific Abs against Mycobacterium bovis in pigs or wild boars, but not in ruminants. The objective of this study was, therefore, to evaluate the performance of an in house-ELISA for TB diagnosis (P22 ELISA) in goats as an experimental model for the diagnosis of TB using oral fluid samples. Oral fluid samples from 64 goats from a TB-infected herd ( n = 197) and all the animals from a TB-free herd ( n = 113) were analyzed using the P22 ELISA. The estimated sensitivity (Se) and specificity (Sp) were 34.4% (95% CI: 22.4-45.6) and 100% (95% CI: 97.4-100), respectively. The optimal cut-off point was set at 100% according to the ROC analysis. Those animals with a higher level of Abs in their oral fluid attained a higher lesion score ( p = 0.018). In fact, when taking into account only the setting of the animals with severe lesions ( n = 16), the ELISA showed a Se of 75% (95% CI: 53.7-96.2). Results of the present study suggest that the P22 ELISA is highly specific but has a limited value detecting infected animals in oral fluid samples. Nevertheless, its performance is significantly higher in the presence of severe lesions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ortega, Infantes-Lorenzo, Bezos, Roy, de Juan, Romero, Moreno, Gómez-Buendía, Agulló-Ros, Domínguez and Domínguez.)

Background: Animal tuberculosis (TB) is distributed worldwide and has a wide range of wild and domestic reservoirs. Few studies concerning TB in camelids have been published in the last decade, particularly as regards Old World Camelids (OWC), but the increase in reports of TB outbreaks in these species in recent years suggests a high susceptibility to the infection., Case Presentation: We studied a dromedary camel (Camelus dromedarius) herd (n = 24) in which a Mycobacterium caprae infection was detected. The TB infection was confirmed in one animal at necropsy through the detection of TB lesions, mainly in the abdominal organs, and the subsequent isolation of M. caprae (SB0157 spoligotype). The whole herd was additionally tested using cellular and humoral based diagnostic techniques. The intradermal tuberculin test results were compared with those obtained using P22 ELISA for the detection of specific antibodies against the M. tuberculosis complex. The TB infected animal was a positive reactor to both the intradermal tuberculin tests and P22 ELISA, while the others were negative to all the diagnostic tests., Conclusion: The present study found M. caprae infection in OWC. This is the first report of M. caprae infection in an OWC not living in a zoo. Since the animal was born in the herd and fed with goat's milk, this practice was suspected to be the potential source of TB infection, which was not confirmed in the other animals present in the herd. Moreover, our results highlight that the intradermal tuberculin test and the P22 ELISA could be valuable tools for the diagnosis of TB in OWC.

Tuberculosis (TB) in small ruminants is a neglected disease despite its major impact on goat and sheep production and the global public health. The awareness of the role of small ruminants in the epidemiology of animal TB has increased in the last two decades, however, there is a lack of standardization of procedures and robust quantitative estimates on the accuracy of diagnostic TB tests in the scientific literature. To address this knowledge gap, all the available information regarding the use of ante-mortem diagnostic techniques in small ruminants was collected and summarized through a systematic review process. Furthermore, a random-effects meta-analysis was conducted to separately estimate the sensitivity (Se) and specificity (Sp) of cell-based tests among the retrieved studies in goats. Studies included in the meta-analysis were also evaluated using the Quality Assessment of Diagnostic Accuracy Studies included in systematic reviews adapted for animal diagnostic tests (VETQUADAS). Median pooled Se estimates of the single intradermal tuberculin (SIT) test (ranged from 0.51 to 0.59), the comparative intradermal tuberculin (CIT) test (ranged from 0.30 to 0.50) and the interferon-gamma (IFN-γ) release assay (IGRA) (ranged from 0.66 to 0.72) were lower than that reported previously in cattle, regardless the interpretation criteria and the reporting of MAP infection or vaccination. However, the specificity was adequate for all the tests (ranged from 0.95 to 0.99), except for the SIT test in MAP vaccinated herds (ranged from 0.78 to 0.90). This study provides an overview of the accuracy of diagnostic tests for TB in goats, however, the considerable between-study heterogeneity found hampered the conclusive interpretation of the pooled Se and Sp estimates. Therefore, further studies in small ruminants are necessary to optimize the diagnostic Se, which could help to design effective control strategies, accelerate the eradication of TB in these species and harmonize test procedures., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Human adenovirus (HAdV) infection has an important clinical impact in the immunosuppressed population and is associated with high morbidity and mortality rates. The lack of a specific, safe and effective antiviral treatment against HAdV makes necessary the search for new therapeutic options. The aim of this study was to evaluate the in vitro activity of ganciclovir (GCV) against HAdV in co-infection by human cytomegalovirus (HCMV) and HAdV in cellular cultures. Quantitative real-time polymerase chain reaction (qPCR) was used to measure HAdV and HCMV DNA replication efficiency in monocultures and in co-infection situations in the presence of both cidofovir (CDV) and GCV. The effects of GCV and CDV were also evaluated in a burst assay (used to measure the production of virus particles) for both viruses, alone and in combination. GCV decreased by 1-log the HAdV DNA replication efficiency in co-infection with HCMV compared with its activity in HAdV monoculture. The burst assay showed that the reductions in virus yield in the presence of GCV were higher for HCMV and co-infection than for HAdV in monoculture (145.2±35.5- vs. 116.4±27.3- vs. 23.0±10.0-fold, respectively, P<0.05). The improved anti-HAdV activity of GCV during co-infection may be because of the more efficient phosphorylation of GCV by the HCMV protein kinase, UL97. Patients treated with GCV as pre-emptive therapy for HCMV infection may be considered as low-risk for developing HAdV infections; however, further evaluations are required to confirm these results., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Current eradication strategies of tuberculosis (TB) in goats mainly rely on the single intradermal tuberculin test (SIT) and single intradermal cervical comparative tuberculin tests (SICCTs). TB vaccination has been proposed as a cost-effective option in high-prevalence herds or countries where economic compensation for the slaughter of positive animals is not affordable. However, TB vaccination compromises the efficiency of tuberculin-based diagnostic tests. In this study, the performance of a new diagnostic platform, based on the P22 antigenic complex, was assessed for skin test (ST), interferon-gamma release assay (IGRA), and serology under different TB scenarios. The sensitivity (Se) of diagnostic tests was assessed in TB-infected goats from the same farm (herd A, N = 77). The specificity (Sp) was assessed in two TB-negative farms (both vaccinated against paratuberculosis): one TB unvaccinated (herd B, N = 77) and another vaccinated with bacille Calmette-Guérin (BCG) (herd C, N = 68). The single (s) P22-IGRA showed the highest Se among IGRA tests (91%), and the comparative (c) P22-ST showed the highest Sp (100% in herd B and 98% in herd C). Combined interpretation of techniques enabled the best diagnostic performances. Combining the SICCT + sP22-IGRA improved Se (97%) compared to SICCT + tuberculin-based IGRA (95%), with a reduction of Sp (95 and 100%, respectively). Besides, combination of P22-ELISA with cP22-ST or SICCT elicited a similar performance in the non-vaccination context (Se: 94 and 95%; Sp: 95 and 95%, respectively), but Sp was significantly higher for the combination with cP22-ST compared to SICCT in the TB vaccination context (95 and 79%, respectively). The combination of serological tests based on P22 and MPB83 showed higher complementarity and improved 13 percentage points the Se of P22-ELISA alone. These findings suggest that either cell-mediated or antibody-based diagnostic techniques, using the P22 antigen complex, can contribute to improve the immunodiagnostics of TB in goats under different TB control strategies., (Copyright © 2020 Arrieta-Villegas, Infantes-Lorenzo, Bezos, Grasa, Vidal, Mercader, Singh, Domingo, de Juan and Pérez de Val.)

Attaining and maintaining the Official Tuberculosis Free status continues to be a challenge when several domestic and wild hosts contribute to the maintenance of the Mycobacterium tuberculosis complex (MTC). Local tuberculosis hotspots are sometimes identified in cattle in low-prevalence regions. We have, therefore, studied one such hotspot in depth in order to produce an epidemiological diagnosis. Host population size and MTC prevalence were estimated in selected wildlife and in livestock, while on-cattle environmental DNA detection was additionally used as a proxy for risk of exposure at the farm (herd) level. Positive skin test reactors were found on16 of the 24 cattle farms studied in the period 2012-2016. Although all goats tested negative to the skin test during this period, MTC was confirmed in four sheep at slaughter, thus indicating an unknown prevalence of infection in this host species. With regard to wildlife, the prevalence of MTC infection based on culture was 8.8% in the case of wild boar (Sus scrofa), and the only road-killed badger (Meles meles) submitted for culture tested positive. Two criteria were employed to divide the cattle farms into higher or lower risk: tuberculosis testing results and environmental DNA detection. Environmental MTC DNA detection yielded significant differences regarding "use of regional pastures" and "proximity to woodland". This study suggests that on-animal environmental DNA sampling may help when assessing contact risk as regards MTC in livestock at the herd level. This tool opens up new avenues of epidemiological research in complex multi-host settings., Competing Interests: I have read the journal's policy and the authors of this manuscript have the following competing interests: MB and SABIOTEC are consultants and offer risk assessment at the wildlife-livestock interface. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Recent studies show that sheep could be considered to be a maintenance host for the causative agents of animal tuberculosis (TB). The performance of diagnostic tests is not well established, and new tests need to be developed for this species. In addition, information about TB prevalence in sheep is scarce. Our objectives were to evaluate a new P22 ELISA for detection of specific antibodies against Mycobacterium tuberculosis Complex (MTC), and to assess the seropositivity in 3998 sheep from herds sampled in TB hotspot areas of northern Atlantic Spain with a low TB prevalence in cattle. Results based on 80 sheep of known infection status suggest excellent sensitivity and specificity (100% and 98%, respectively) even in a M. avium susbsp. paratuberculosis infected flock. The observed TB seroprevalence was 17.96% (698/3998; CI95% 16.31-18.67). Our results indicate that the P22 ELISA may constitute a good option for TB screening at the herd level in sheep, and that sheep are an important host and control programs should be implemented at least in hotspots or when cohabiting with other TB-infected species, i.e. cattle and goats., Competing Interests: Declaration of Competing Interest None of the authors of this paper has a financial or personal relationship with other people or organizations that could inappropriately influence or bias the content of the paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Osteoclasts (OC) originate from either bone marrow (BM)-resident or circulating myeloid OC progenitors (OCP) expressing the receptor CX 3 CR1. Multiple lines of evidence argue that OCP in homeostasis and inflammation differ. We investigated the relative contributions of BM-resident and circulating OCP to osteoclastogenesis during homeostasis and fracture repair. Using CX 3 CR1-EGFP/TRAP tdTomato mice, we found CX 3 CR1 expression in mononuclear cells, but not in multinucleated TRAP + OC. However, CX 3 CR1 - expressing cells generated TRAP + OC on bone within 5 d in CX 3 CR1CreERT2/Ai14 tdTomato reporter mice. To define the role that circulating cells play in osteoclastogenesis during homeostasis, we parabiosed TRAP tdTomato mice (CD45.2) on a C57BL/6 background with wild-type (WT) mice (CD45.1). Flow cytometry (CD45.1/45.2) demonstrated abundant blood cell mixing between parabionts after 2 wk. At 4 wk, there were numerous tdTomato + OC in the femurs of TRAP tdTomato mice but almost none in WT mice. Similarly, cultured BM stimulated to form OC demonstrated multiple fluorescent OC in cell cultures from TRAP tdTomato mice, but not from WT mice. Finally, flow cytometry confirmed low-level engraftment of BM cells between parabionts but significant engraftment in the spleens. In contrast, during fracture repair, we found that circulating CX 3 CR1 + cells migrated to bone, lost expression of CX 3 CR1, and became OC. These data demonstrate that OCP, but not mature OC, express CX 3 CR1 during both homeostasis and fracture repair. We conclude that, in homeostasis mature OC derive predominantly from BM-resident OCP, whereas during fracture repair, circulating CX 3 CR1 + cells can become OC., (Copyright © 2020 by The American Association of Immunologists, Inc.)

In Europe, badgers ( Meles meles ) are recognized as major tuberculosis (TB) reservoir hosts with the potential to transmit infection to associated cattle herds. Recent studies in Spain have demonstrated that vaccination with a heat-inactivated Mycobacterium bovis vaccine (HIMB) successfully protects captive wild boar and red deer against progressive disease. The aim of this study was to evaluate the efficacy of two oral vaccines against TB in a badger model: the live-attenuated M. bovis bacillus Calmette-Guérin BCG vaccine (Danish strain) and a HIMB vaccine. Twenty-four badgers were separated in three treatment groups: oral vaccinated with live BCG (10 8 CFU, n = 5), oral vaccinated with HIMB (10 7 CFU , n = 7), and unvaccinated controls ( n = 12). All badgers were experimentally infected with M. bovis (10 3 CFU) by the endobronchial route targeting the right middle lung lobe. Throughout the study, clinical, immunological, pathological, and bacteriological parameters of infection were measured. Both vaccines conferred protection against experimental TB in badger, as measured by a reduction of the severity and lesion volumes. Based on these data, HIMB vaccination appears to be a promising TB oral vaccine candidate for badgers in endemic countries., (Copyright © 2020 Balseiro, Prieto, Álvarez, Lesellier, Davé, Salguero, Sevilla, Infantes-Lorenzo, Garrido, Adriaensen, Juste and Barral.)

Caprine tuberculosis (TB) is a zoonosis with sanitary and economic repercussions. Caprine TB control programs are based on a test and cull strategy using the intradermal tuberculin tests and slaughterhouse surveillance. However, this approach is not always feasible and may have a limited sensitivity under specific circumstances. In this study, performance of a new experimental test based on the P22 protein complex (P22 ELISA) was evaluated in two TB-infected herds using milk and serum samples and compared with cell-based diagnostic tests. Samples from a low (n = 62, herd 1) and a high (n = 52, herd 2) TB prevalence herd were selected. Moreover, bulk tank milk samples from both herds were analysed using the P22 ELISA. At the end of the study, a group of animals (n = 21) was euthanized and subjected to post-mortem analysis and bacteriological culture. Significant differences (p < .001) on the qualitative and quantitative (ODs) results were observed between herds using both serum and milk samples in the P22 ELISA. The correlation observed in the quantitative results obtained in serum and milk samples was very strong in animals from flock 2 (r s  = 0.91) and moderate in animals from flock 1 (r s  = 0.46). Among the slaughtered animals, the P22 ELISA detected a higher proportion of lesion-culture positive animals than cell-based diagnostic tests (61.9 and 66.7% using milk and serum samples, respectively). The P22 ELISA using milk samples demonstrated a similar sensitivity compared with serum samples, suggesting it might be a valuable test for TB control in dairy goats., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

In recent years, human adenovirus (HAdV) infections have shown a high clinical impact in both immunosuppressed and immunocompetent patients. The research into specific antiviral drugs for the treatment of HAdV infections in immunocompromised patients constitutes a principal objective for medicinal chemistry due to the lack of any specific secure drug to treat these infections. In this study, we report a small-molecule library (67 compounds) designed from an optimization process of piperazine-derived urea privileged structures and their biological evaluation: antiviral activity and cytotoxicity. The active compounds selected were further evaluated to gain mechanistic understanding for their inhibition. Twelve derivatives were identified that inhibited HAdV infections at nanomolar and low micromolar concentrations (IC 50 from 0.6 to 5.1 μM) with low cytotoxicity. In addition, our mechanistic assays suggested differences in the way the derivatives exert their anti-HAdV activity targeting transcription, DNA replication and later steps in the HAdV replication cycle. Furthermore, eight of the 12 studied derivatives blocked human cytomegalovirus (HCMV) DNA replication at low micromolar concentrations. The data provided herein indicates that the 12 thiourea/urea piperazine derivatives studied may represent potential lead compounds for clinical evaluation and development of new anti-HAdV drugs., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Pyle's disease (OMIN number 265900) is a metaphyseal dysplasia of benign course, inherited with an autosomal recessive pattern. Some 30 genuine cases have been described so far. The cause of this process has been known since 2016, when its relationship to mutations in the gene encoding the sFRP protein, a known inhibitor of the Wnt pathway, was discovered. We report the case of a 58-year-old man, diagnosed with Pyle's disease based on his clinical and radiographic characteristics, whose phenotype suggested a differential control of cortical and trabecular bone homeostasis., (Copyright © 2018 Elsevier España, S.L.U. and Sociedad Española de Reumatología y Colegio Mexicano de Reumatología. All rights reserved.)

: We provide a temporal overview (from 2012 to 2018) of the outcomes of tuberculosis (TB) in the cattle and badger populations in a hot-spot in Asturias (Atlantic Spain). We also study the badger's spatial ecology from an epidemiological perspective in order to describe hazardous behavior in relation to TB transmission between cattle and badgers. Culture and single intradermal tuberculin test (SITT) were available for cattle as part of the National Program for the Eradication of TB. A field survey was also carried out in order to determine the paddocks and buildings used by each farm, and the information obtained was stored by using geographic information systems. Moreover, eighty-three badgers were submitted for necropsy and subsequent bacteriological studies. Ten badgers were also tracked, using global positioning system (GPS) collars. The prevalence of TB in cattle herds in the hot-spot increased from 2.2% in 2012 to 20% in 2016; it then declined to 0.0% in 2018. In contrast, the TB prevalence in badgers increased notably (from 5.55% in 2012-2015 to 10.64% in 2016-2018). Both cattle and badgers shared the same strain of Mycobacterium bovis . The collared badgers preferred paddocks used by TB-positive herds in spring and summer (when they were more active). The males occupied larger home ranges than the females (Khr95: males 149.78 ± 25.84 ha and females 73.37 ± 22.91 ha; Kcr50: males 29.83 ± 5.69 ha and females 13.59 ± 5.00 ha), and the home ranges were smaller in autumn and winter than in summer. The averages of the index of daily and maximum distances traveled by badgers were 1.88 ± (SD) 1.20 km and 1.99 ± 0.71 km, respectively. One of them presented a dispersive behavior with a maximum range of 18.3 km. The most preferred habitat was apple orchards in all seasons, with the exception of winter, in which they preferred pastures. Land uses and landscape structure, which have been linked with certain livestock-management practices, provide a scenario of great potential for badger-cattle interactions, thus enhancing the importance of the badgers' ecology, which could potentially transmit TB back to cattle in the future., Competing Interests: The authors declare no conflicts of interest.

Effective vaccines against tuberculosis (TB) are needed in order to prevent TB transmission in human and animal populations. Evaluation of TB vaccines may be facilitated by using reliable animal models that mimic host pathophysiology and natural transmission of the disease as closely as possible. In this study, we evaluated the immunogenicity and efficacy of two attenuated vaccines, BCG and MTBVAC, after each was given to 17 goats (2 months old) and then exposed for 9 months to goats infected with M. caprae. In general, MTBVAC-vaccinated goats showed higher interferon-gamma release than BCG vaccinated goats in response to bovine protein purified derivative and ESAT-6/CFP-10 antigens and the response was significantly higher than that observed in the control group until challenge. All animals showed lesions consistent with TB at the end of the study. Goats that received either vaccine showed significantly lower scores for pulmonary lymph nodes and total lesions than unvaccinated controls. Both MTBVAC and BCG vaccines proved to be immunogenic and effective in reducing severity of TB pathology caused by M. caprae. Our model system of natural TB transmission may be useful for evaluating and optimizing vaccines.

We found that protease-activated receptor 1 (PAR1) was transiently induced in cultured osteoclast precursor cells. Therefore, we examined the bone phenotype and response to resorptive stimuli of PAR1-deficient (knockout [KO]) mice. Bones and bone marrow-derived cells from PAR1 KO and wild-type (WT) mice were assessed using microcomputed tomography, histomorphometry, in vitro cultures, and RT-PCR. Osteoclastic responses to TNF-α (TNF) challenge in calvaria were analyzed with and without a specific neutralizing Ab to the Notch2-negative regulatory region (N2-NRR Ab). In vivo under homeostatic conditions, there were minimal differences in bone mass or bone cells between PAR1 KO and WT mice. However, PAR1 KO myeloid cells demonstrated enhanced osteoclastogenesis in response to receptor activator of NF-κB ligand (RANKL) or the combination of RANKL and TNF. Strikingly, in vivo osteoclastogenic responses of PAR1 KO mice to TNF were markedly enhanced. We found that N2-NRR Ab reduced TNF-induced osteoclastogenesis in PAR1 KO mice to WT levels without affecting WT responses. Similarly, in vitro N2-NRR Ab reduced RANKL-induced osteoclastogenesis in PAR1 KO cells to WT levels without altering WT responses. We conclude that PAR1 functions to limit Notch2 signaling in responses to RANKL and TNF and moderates osteoclastogenic response to these cytokines. This effect appears, at least in part, to be cell autonomous because enhanced osteoclastogenesis was seen in highly purified PAR1 KO osteoclast precursor cells. It is likely that this pathway is involved in regulating the response of bone to diseases associated with inflammatory signals., (Copyright © 2019 by The American Association of Immunologists, Inc.)

Background: Serum antibody detection has potential as a complementary diagnostic tool in animal tuberculosis (TB) control, particularly in multi-host systems. The objective of the present study was to assess the specificity (Sp) of an enzyme-linked immunosorbent assay (ELISA) based on the new multiprotein complex P22 for the detection of specific antibodies against the Mycobacterium tuberculosis complex (MTC) in the four most relevant domestic animals acting as MTC hosts: cattle, goat, sheep and pig. We used sera from an officially TB-free (OTF) country, Norway, and from a non-OTF one, Spain. The samples included sera from goats that had been vaccinated against M. avium subsp. paratuberculosis (MAP) and sheep from a herd in which Corynebacterium pseudotuberculosis had been isolated., Results: In cattle, the Sp ranged from 92.5 (IC95% 90.7-94) to 99.4% (IC95% 98.3-99.8) depending on the cut-off used and the origin of the samples (Spain or Norway). Sp in cattle (cut-off point 100) was significantly higher (P < 0.05) for Norwegian samples. By contrast, Sp in goats was consistently low at the 100 cut-off [30.9 (CI95%23.4-39.5)-78% (CI95% 68.9-85)]. A higher cut-off of 150 improved Sp in Norwegian goats [97% (CI95% 91.6-99)], but still yielded a poor Sp of 56.1% (CI95% 47.3-64.6) in Spanish goats. In Norway at the 100 cut-off the Sp was 58.3 (CI95% 42.2-72.9) and 90.6% (CI95% 81-95.6) in MAP vaccinated and non-vaccinated goats, respectively, indicating interference due to MAP vaccination. Sp in sheep was between 94.4 (CI95% 91.7-96.3) and 100% (CI95% 96.3-100) depending on the cut-off and country, and no diagnostic interference due to infection with C. pseudotuberculosis was recorded. Sp in pigs was 100%, regardless the cut-off point applied, and no significant differences were observed between pigs from Norway and from Spain., Conclusions: Due to its excellent Sp in pigs and acceptable Sp in cattle and sheep, this ELISA may constitute a suitable option for TB screening at herd level, particularly in OTF-countries.

European badgers (Meles meles) have been identified as wildlife reservoirs for Mycobacterium bovis in the UK and Ireland, and may also have a role in the epidemiology of animal tuberculosis in other European regions. Thus, detection of M. bovis-infected badgers may be required for the purposes of surveillance and monitoring of disease levels in infected populations. Current serological assays to detect M. bovis infection in live badgers, while rapid and inexpensive, show limited diagnostic sensitivity. Here we describe and evaluate new ELISA platforms for the recognition of the P22 multiprotein complex derived from the purified protein derivative (PPD) of M. bovis. The recognition of IgG against P22 multiprotein complex derived from PPD-B was tested by ELISA in the serum of badgers from the UK, Ireland and Spain. TB infection in the badgers was indicated by the presence of M. bovis in tissues by culture and histology at post-mortem examination and TB-free status was established by repeated negativity in the interferon γ release assay (IGRA). In experimentally infected badgers, humoral antibody responses against P22 developed within 45 days post-infection. The ELISA tests showed estimated sensitivity levels of 74-82% in experimentally and naturally infected badgers with specificities ranging from 75% to 100% depending on the badger population tested. The P22 multi-antigen based ELISAs provide a sensitive and specific test platform for improved tuberculosis surveillance in badgers., (© 2019 Crown copyright. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

To study complement function in mammalian leishmanioses, we developed mouse monoclonal antibodies to the human complement system components C1q, C4, factor D, factor H, factor B, properdin, C5 and C9. Antibody specificity was determined by indirect and capture ELISA and by Western blot. In flow cytometry analysis, seven antibodies recognized the cognate component on human serum-opsonized Leishmania promastigotes. Antibody reactivity was screened against promastigotes opsonized with sera of nine mammalian genera: pig, guinea pig, goat, rabbit, cat, dog, hamster, jird and rat. No antibody recognized jird epitopes on promastigotes. Anti-C4, -properdin, and -C5b reacted with the orthologous protein of all other mammals tested except cat (anti-properdin) and hamster (anti-C5b); anti-C9 only recognized the rabbit ortholog, and anti-C1q, -factor B and -factor H did not react with any of the nine orthologs. Such interspecies crossreactive antibodies can be valuable tools for analysis of mammalian complement function in infectious diseases., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Red deer (Cervus elaphus) farming is a growing economic activity worldwide. However, the capacity of this species to act as reservoir of animal tuberculosis (TB) poses a threat to other wildlife and to livestock. Diagnostic assay accuracy in this species is therefore highly relevant for prevention and control measures. Our aim was to evaluate the diagnostic performance of the protein complex P22, obtained from Mycobacterium bovis derived purified protein derivative (bPPD), as a candidate antigen for the detection of antibodies against Mycobacterium tuberculosis complex (MTC). We assessed the performance of this new antigen in indirect enzyme-linked immunosorbent assays (ELISA) in TB-positive and TB-negative red deer, in comparison with a bPPD-based ELISA. The P22 ELISA achieved a higher specificity (Sp) and similar sensitivity (Se) in comparison with the bPPD ELISA at all the cut-off points considered. The P22 ELISA yielded optimal Sp (99.02%; 95% confidence intervals [CI 95% ]: 96.5-99.8) and appropriate Se (70.1%; CI 95% : 63.6-76) at the selected cut-off point of 100%. These results suggest that P22 can be used as an alternative antigen in the immunodiagnosis of animal TB through the use of an ELISA-type detection of antibodies against MTC in red deer, thus contributing to the diagnosis of animal TB in this species as a measure for further disease prevention and control programs., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The repositioning of drugs already approved by regulatory agencies for other indications is an emerging alternative for the development of new antimicrobial therapies. The repositioning process involves lower risks and costs than the de novo development of novel antimicrobial drugs. Currently, infections by adenovirus show a steady increment with a high clinical impact in immunosuppressed and immunocompetent patients. The lack of a safe and efficacious drug to treat these infections supports the search for new antiviral drugs. Here we evaluated the anti-adenovirus activity of niclosanide, oxyclozanide, and rafoxanide, three salicylanilide anthelmintic drugs. Also, we carried out the cytotoxicity evaluation and partial characterization of the mechanism of action of these drugs. The salicylanilide anthelmintic drugs showed significant anti-adenovirus activity at low micromolar concentrations with little cytotoxicity. Moreover, our mechanistic assays suggest differences in the way the drugs exert anti-adenovirus activity. Niclosamide and rafoxanide target transport of the HAdV particle from the endosome to the nuclear envelope, whilst oxyclozanide specifically targets adenovirus immediately early gene E1A transcription. Data suggests that the studied salicylanilide anthelmintic drugs could be suitable for further clinical evaluation for the development of new antiviral drugs to treat infections by adenovirus in immunosuppressed patients and in immunocompetent individuals with community-acquired pneumonia.