LabVIEW-based control and acquisition system for the dosimetric characterization of a silicon strip detector

Theaimofthisworkistopresentanewdataacquisition,control,andanalysissoftwaresystemwrittenin LabVIEW.Thissystemhasbeendesignedtoobtainthedosimetryofasiliconstripdetectorinpolyethylene. It allows the full automation of the experiments and data analysis required for the dosimetric characterization of silicon detectors. It becomes a useful tool that can be applied in the daily routine check of a beam accelerator.MINECO ICTI2013-2016/FPA2013-47327-C2-1-RMINECO ICTI2013-2016/FPA2014-53290-C2-2- PJunta de Andalucía P12-FQM-160

Identificación y caracterización molecular del Norovirus circulante en la ganadería porcina española

Norovirus es un agente patógeno que causa la mayoría de gastroenteritis de origen no bacteriano en humanos a nivel mundial. Es capaz de infectar a un gran rango de hospedadores aparte del ser humano. Entre los Norovirus animales, el de porcino (genogrupo II) es el que más se asemeja genéticamente al Norovirus que afecta al ser humano. Así mismo, España representa uno de los principales países productores de porcino en Europa y a nivel mundial. A pesar de la relevancia del sector porcino y el posible potencial zoonótico del Norovirus que afecta a esta especie, existe poca información sobre la circulación de Norovirus porcino en nuestro país. El objetivo de este trabajo es identificar y evaluar la incidencia de este virus en la cabaña porcina española mediante el análisis de 480 muestras clínicas recibidas por Exopol S.L. durante un período de tres años (2020-2022) y analizar las relaciones filogenéticas entre estas cepas y otras previamente descritas en porcinos y humanos.Para la identificación de Norovirus porcino en diferentes tipos de muestras biológicas, se diseñó un ensayo de RT-PCR en tiempo real que presentó una mayor sensibilidad que otro ensayo previamente descrito. Mediante esta metodología, Norovirus fue identificado en un 10,8 % (52/480) de las muestras clínicas del estudio. La frecuencia de detección fue mayor en las muestras de cerdo ibérico y parece asociarse al sistema de producción característico de esta raza autóctona. La circulación de Norovirus en cerdo blanco resultó particularmente alta en el caso de cerdos de cebo (11,5 %), al igual que en estudios similares.Este trabajo describe la primera caracterización molecular de Norovirus porcino en España, identificando dos genotipos de cepas diferentes (GII.18 y GII.11) y que corroboran los resultados de estudios previos de genotipado realizados para este agente en otros países europeos. Por otro lado, el análisis filogenético demostró una gran variabilidad entre los dos genotipos identificados, y plantea la necesidad de hacer posteriores estudios con el fin de evaluar mejor el origen de estas cepas, así como un seguimiento para rastrear la aparición de virus potencialmente zoonóticos derivados de eventos de recombinación. Por tanto, este trabajo resulta un esfuerzo por poner en práctica el concepto “One Health”.<br /

Accurate Diagnosis of Small Ruminant Lentivirus Infection Is Needed for Selection of Resistant Sheep through TMEM154 E35K Genotyping

[EN] Small ruminant lentiviruses (SRLV) cause an incurable multiorganic disease widely spread in sheep and goats that disturbs animal welfare and production. In the absence of a vaccine, control measures have been traditionally based on early diagnosis and breeding with virus-inactivated colostrum with segregation of seropositive animals. However, antigenic heterogeneity, poor antibody production due to low viral load, and single strain design of most available ELISA, pose a threat to SRLV diagnosis. Genome-wide association studies have described TMEM154 E35K polymorphism as a good genetic marker for selection of resistant animals in some American and European breeds. In this study, a multitargeted serological and virological screening of more than 500 animals from four different breeds (latxa, raza Navarra, assaf, and churra) attending to SRLV infection status was performed. Then, animals were genotyped to characterize TMEM154 E35K polymorphism. ELISA procedures, individually considered, only identified a proportion of the seropositive animals, and PCR detected a fraction of seronegative animals, globally offering different animal classifications according to SRLV infection status. TMEM154 allele frequency differed substantially among breeds and a positive association between seroprevalence and TMEM154 genotype was found only in one breed. Selection based on TMEM154 may be suitable for specific ovine breeds or SRLV strains, however generalization to the whole SRLV genetic spectrum, ovine breeds, or epidemiological situation may need further validationSIThis research was funded by Spanish Ministry of Science, Innovation, and Universities, grant number RTI2018-096172-B-C31. The APC was funded by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI). I.E. was funded by Universidad Pública de Navarra. Hugo Ramírez was financially supported by the “Programa de apoyos para la superación del personal académico (PASPA)” scholarship from DGAPA-UNA

Occurrence of rotavirus a genotypes and other enteric pathogens in diarrheic suckling piglets from Spanish swine farms

Species A rotavirus (RVA) is a major viral pathogen causing diarrhea in suckling piglets. Studies on its genetic heterogeneity have implications for vaccine efficacy in the field. In this study, fecal samples (n = 866) from diarrheic piglets younger than 28 days were analyzed over a two-year period (2018–2019). Samples were submitted from 426 farms located in 36 provinces throughout Spain and were tested using real-time PCR (qPCR) and reverse transcription real-time PCR (RT-qPCR) for five enteric pathogens. The individual prevalence was 89.4%, 64.4%, 44.9%, 33.7% and 4.4% for Clostridiumperfringens, Clostridioides (formerly Clostridium) difficile, species A rotavirus, species C rotavirus and porcine epidemic diarrhea virus, respectively. Most specimens (96.9%) were positive for at least one of the target pathogens, and more than 80% of samples harbored mixed infections. Nucleotide sequencing of 70 specimens positive for RVA revealed the presence of the VP7 genotypes G4, G9, G3, G5, G11 and the VP4 genotypes P7, P23, P6 and P13, with the combinations G4P7 and G9P23 being the most prevalent, and especially in the areas with the highest pig population. The study shows the extensive genetic diversity of RVA strains as well as discrepancies with the genotypes contained in the vaccine available in Spain, and multiple amino acid differences in antigenic epitopes of different G- and P- genotypes with the vaccine strains. Further investigations are needed to determine the efficacy of the vaccine to confer clinical protection against heterologous strains

Identificación molecular del subtipo del virus de Influenza A circulante en la cabaña de porcinos en España mediante PCR en tiempo real y secuenciación

El virus de la Influenza A es el patógeno responsable de la gripe porcina (SIV), una enfermedad respiratoria con alta prevalencia en las explotaciones en nuestro país y en donde ocasiona importantes pérdidas económicas. Durante las últimas décadas, se han descritoprincipalmente cuatro subtipos virales de SIV (H1avN1, H1huN2, H3N2 y H1N1pdm), circulando entre la población porcina europea. Sin embargo, la rápida evolución de estos agentes ha dado lugar a la aparición de nuevos subtipos por procesos de recombinación genética característicos de estos virus.La realización del presente estudio ha permitido identificar mediante RT-qPCR, el subtipo del 94% (124/132) de casos clínicos positivos a SIV procedentes de granjas españolas y recibidos en el laboratorio Exopol durante el año 2020. Entre los subtipos identificados, el H1avN2 (43%) fue el más frecuente, un resultado similar a lo descrito por Sosa Portugal y col.en años previos en porcinos de la península ibérica. En otros países europeos, sin embargo, el subtipo H1avN1 sigue siendo el subtipo más extendido. El resto de subtipos identificados en nuestro trabajo fueron los siguientes: H1huN2 (16%), H1avN1 (10%), H3N1 (11%), H3N2 (10%), y H1pdmN1pdm (2%). La diversidad genética de los SIV identificados en el presente estudio fue evaluada mediante secuenciación Sanger de los genes HA y NA. Un total de 29 secuencias para el gen HA y 36 secuencias para el gen NA fueron obtenidas de casos clínicos procedentes de distintas provincias españolas. El análisis filogenético de las secuencias demostró una mayor similitud con secuencias de cepas españolas descritas en años previos que con cepas de SIV originarias de otros países europeos. Además, el gen NA se encuentra generalmente más conservado que el gen HA, aunque en este caso, las secuencias de tipo H3 presentaron la mayor similitudnucleotídica. La vigilancia continua de los SIV es de gran importancia para la industria porcina, tanto para monitorear la evolución del virus que cambia rápidamente, como para conocer la situación epidemiológica de una región o un país. Es por estos motivos que los datos obtenidos en este trabajo no sólo nos han permitido confirmar la circulación de las tres cepas clásicas porcinas, sino que también han constituido la primera descripción de una nueva cepa H1pdmN1 en la cabaña porcina española.<br /

Molecular diagnosis of footrot and contagious ovine digital dermatitis in small ruminants in the Iberian Peninsula

Contagious ovine digital dermatitis (CODD) and footrot (FR), a sub-acute or acute necrotic (decaying) infectious disease involving the hoof and underlying tissues, pose economic challenges to herds in Spain and worldwide. The aetiological agent for FR is Dichelobacter nodosus, while CODD is caused by pathogenic Treponema phylogroups. We detail the findings derived from the analysis by qPCR of 105 pooled samples from 100 ovine and five caprine herds in Spain and Portugal, alongside 15 samples from healthy flocks in order to identify Dichelobacter nodosus, Fusobacterium necrophorum, Treponema spp., and three pathogenic Treponema phylogroups (T. phagedenis, T. medium, and T. pedis). Treponema spp. were detected in all 120 pools, including samples from the 15 healthy flocks where only one positive result for F. necrophorum was recorded. Mixed infections by agents different from Treponema spp. were identified in 68.57% of samples. Positive results for F. necrophorum and/or D. nodosus, were obtained for 91.4% of the pools, whereas the presence of the three pathogenic Treponema phylogroups was rare: each of them appeared in isolation in a single pool, while they were found in 18 pools in combination with other agents. While F. necrophorum was the sole finding in 16.2% of samples from affected herds, D. nodosus (the footrot causative agent) was only detected in 61% of affected farms. An improved qPCR protocol was implemented to determine the serogroups of D. nodosus in the samples and found all of them (except the G serogroup), often in combined infections (35.1%). This report concludes with comprehensive proposals for diagnosing, preventing, and treating hoof ailments, remarking the interest of the information about D. nodosus serogroups in order to improve the efficiency of immunization by choosing appropriate vaccine protocols

Nurr1 protein is required for N-Methyl-d-aspartic Acid (NMDA) receptor-mediated neuronal survival

NMDA receptor (NMDAR) stimulation promotes neuronal survival during brain development. Cerebellar granule cells (CGCs) need NMDAR stimulation to survive and develop. These neurons differentiate and mature during its migration from the external granular layer to the internal granular layer, and lack of excitatory inputs triggers their apoptotic death. It is possible to mimic this process in vitro by culturing CGCs in low KCl concentrations (5 mm) in the presence or absence of NMDA. Using this experimental approach, we have obtained whole genome expression profiles after 3 and 8 h of NMDA addition to identify genes involved in NMDA-mediated survival of CGCs. One of the identified genes was Nurr1, a member of the orphan nuclear receptor subfamily Nr4a. Our results report a direct regulation of Nurr1 by CREB after NMDAR stimulation. ChIP assay confirmed CREB binding to Nurr1 promoter, whereas CREB shRNA blocked NMDA-mediated increase in Nurr1 expression. Moreover, we show that Nurr1 is important for NMDAR survival effect. We show that Nurr1 binds to Bdnf promoter IV and that silencing Nurr1 by shRNA leads to a decrease in brain-derived neurotrophic factor (BDNF) protein levels and a reduction of NMDA neuroprotective effect. Also, we report that Nurr1 and BDNF show a similar expression pattern during postnatal cerebellar development. Thus, we conclude that Nurr1 is a downstream target of CREB and that it is responsible for the NMDA-mediated increase in BDNF, which is necessary for the NMDA-mediated prosurvival effect on neurons

Las cuevas de la Sierra de Atapuerca y el uso humano del paisaje kárstico durante el Pleistoceno (Burgos, España)

El karst de la Sierra de Atapuerca representa un interesante sistema multinivel, inactivo y heredado de antiguos niveles de base plio-pleistocenos, que alberga los enclaves prehistóricos más importantes para el conocimiento del poblamiento antiguo en Eurasia, y que fue declarado Patrimonio de la Humanidad en 2000 por la UNESCO. Estas cuevas se originan a partir de conductos subhorizontales con paleodrenajes en sentido SN, localizándose la zona de descarga en la cabecera del río Pico. Los conductos están organizados en tres niveles principales que aparecen colgados entre 90 y 60 m sobre el actual cauce del río Arlanzón, coincidiendo con los niveles de base generados por sus terrazas fluviales T2, T3 y T5. La incisión fluvial liberó de las aguas los conductos superiores mientras se excavaban los niveles inferiores del karst. Las cuevas que iban quedando accesibles fueron utilizadas por la fauna y los homininos, conservando un registro arqueo-paleontológico de más de 1,2 Ma

High Levels of Proinflammatory Cytokines, but Not Markers of Tissue Injury, in Unaffected Intestinal Areas from Patients with IBD

Intestinal alterations in IBD are triggered and maintained by an overexpression of proinflammatory cytokines. Additionally, increased immune activation has been found in the adjacent intestinal areas without displaying any apparent histological alterations, however, the regulatory environment is not well established. Biopsy specimens from patients with ulcerative colitis (UC) and Crohn's disease (CD), from both affected and unaffected areas, and also from a group of colonic biopsies from healthy controls, were included in our study. Cytokines and markers of mucosal damage were analyzed by real-time PCR, and some of the results confirmed by western-blot and ELISA. Levels of IFNγ, TNFα, IL-6, IL-15, IL-18, and IL-23 were increased (above healthy controls) in both affected and unaffected areas from IBD. IL-1β, IL-6, IL-12, and IL-27 were higher in affected areas compared to unaffected ones in UC but not CD. In general, a correlation was observed between mRNA levels of these cytokines and both iNOS and Granzyme B. SOCS-2 and SOCS-3 were also increased in the affected areas. In conclusion, the unaffected areas from IBD show increased levels of a restricted set of cytokines that may exert immune activating roles in these areas without being able to trigger tissue damage

Copper poisoning, a deadly hazard for sheep

Copper (Cu) is an essential microelement for animals. However, sheep are particularly susceptible to Cu intoxication, a deadly disease reported worldwide. The risk of developing this poisoning is higher in vulnerable breeds and in intensively managed lambs or milk sheep. Two types of Cu intoxication can occur depending on the chronic or acute exposure to Cu. In chronic Cu poisoning (CCP), the most common form, Cu is accumulated in the liver during a subclinical period. A low intake of Cu antagonists (molybdenum, sulphur, iron, or zinc) favours Cu accumulation. The sudden release of Cu into the blood causes acute haemolysis with anaemia, haemoglobinuria, jaundice and death within 1–2 days. Acute Cu poisoning is related to the accidental administration or ingestion of toxic amounts of Cu. Acute oral exposure to Cu causes severe gastroenteritis, shock and death. Collapse and death occur shortly after parenteral administration. The diagnosis is based on history, clinical, gross pathological, histological and toxicological findings. Treatment of sheep with severe clinical signs often has poor success but is very effective during the Cu accumulation phase. Different therapies, based on either chelating agents or Cu antagonists, have been used to treat and prevent CCP