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Clostridium difficile es la causa más común de diarrea asociada a hospitales y antimicrobianos en los seres humanos, sin embargo, el papel de C. difficile en perros diarreicos no está definido. Los objetivos del presente trabajo son: Determinar la prevalencia de C. difficile en perros con y sin diarrea y comparar si hay diferencias entre perros hospitalizados y sanos. Analizar si la infección por C. difficile está relacionada con el uso de antibióticos u otros fármacos en perros y determinar la presencia de otros agentes zoonóticos en perros hospitalizados y sanos y su perfil de antibioresistencia. Para realizar este estudio se obtuvieron muestras fecales con un hisopo estéril o bien de heces de 25 perros hospitalizados y 25 perros sanos. Posteriormente las muestras fueron procesadas para realizar diagnóstico microbiológico y diagnóstico molecular. En el aislamiento microbiológico en medio selectivo no se obtuvo ningún aislado de C. difficile en perros hospitalizados ni en perros sanos. Y a partir de la PCR, los resultados fueron negativos en todas las muestras para el gen que codifica la toxina A (tcdA) y el gen que codifica la toxina B (tcdB). Se obtuvieron aislados puros de otros microorganismos zoonóticos (Escherichia coli) y se realizaron antibiogramas para obtener un perfil de resistencia. Se concluye que la prevalencia de colonización de C. difficile es más baja de la esperada, ya que no se ha obtenido ningún resultado positivo de C. difficile ni por aislamiento microbiológico específico ni por detección por PCR de las toxinas tcdA y tcdB. Se ha detectado la presencia de cepas de E. coli multirresistentes en la población canina de la región geográfica estudiada, incluyendo antimicrobianos de uso generalizado en veterinaria y en humana lo que podría suponer un riesgo para la salud pública.

Se describen la sintomatología, alteraciones hematológicas y bioquímicas más importantes, junto con las lesiones histológicas que aparecen en la rinitis crónica de las tortugas terrestres mediterráneas.

FUNDAMENTOS. La hidatidosis o equinococosis es una antropozoonosis parasitaria causada por el género Echinococcus, perteneciente a la familia Taeniidae, posee amplia distribución mundial, siendo endémico en la zona del mediterráneo. Se estima que 2-3 millones de personas en el mundo padecen de esta enfermedad, siendo considerada una de las 20 enfermedades tropicales desatendidas reconocida por la Organización Mundial de la Salud (OMS), también reconocido por las autoridades europeas mediante la directiva 2003/99 / EC y una enfermedad que debe informarse a la Autoridad Europea de Seguridad Alimentaria (EFSA). En el año 1982 la hidatidosis en España fue incluida en la lista de enfermedades de declaración obligatoria. OBJETIVOS. Hay una falta de información por parte de fuentes oficiales españolas como la Red Nacional de Vigilancia Epidemiológica (RENAVE) de la estimación de los casos de hidatidosis importada en España y del país de procedencia de estos. En muchos casos provienen de países de baja renta, de los cuales no se tiene mucha información del estatus sanitario. Por lo tanto este trabajo tiene como objetivo realizar una revisión sistemática de la literatura de casos de hidatidosis importada en España, junto con la recopilación de sus respectivos datos epidemiológicos, y así poder realizar una aproximación de la situación en estos países. MATERIALES Y MÉTODOS. Se realizó una revisión sistemática de la literatura para obtener referencias sobre casos de pacientes inmigrantes diagnosticados con hidatidosis en España. Los artículos debían describir la infección por Echinococcus spp. en al menos una persona inmigrante residiendo en España. RESULTADOS. Se incluyeron 21 artículos publicados desde el año 1995-2018 con un total de 84 casos de hidatidosis importada en España, con una edad promedio de 33.2 años, que van desde los 4 a los 87 años. 34.5% de los casos documentados correspondían al sexo masculino, 23 (27.4%) al género femenino y 32 (38.1%) sin información. África concentró el mayor número de inmigrantes con hidatidosis con una presentación de 39.3% (33), seguido de Europa 22.6% (19) y América del Sur 13.1% (11). El país africano con mayor número de casos de hidatidosis en España corresponde a Marruecos 35.7% (30), en Europa Rumanía con 14.3% (12) y en América del Sur Perú con 9.5% (8). El órgano que más se vio afectado fue el hígado 33.3% (28), seguido de pulmón y corazón con un 4.8% (4) respectivamente. CONCLUSIONES. Con los resultados obtenidos de los 21 artículos recopilados podemos concluir que los pacientes con hidatidosis en España provenían principalmente de Marruecos 35.7% (30), Rumanía 14.3% (12) y Perú con 9.5% (8). Es poca la información de la actual situación de la hidatidosis en estos países, por lo que estos casos pueden actuar como centinela para realizar una estimación de lo que está sucediendo en sus países de origen. Se evidencia falla en los mecanismos de notificación de los casos de hidatidosis en España a través de la Red Nacional de Vigilancia Epidemiológica y la disparidad de información entre este a nivel español y la ECDC/EFSA a nivel Europeo. Por lo que, con esta falta de información es difícil hacer una real estimación de la situación actual de hidatidosis en España y de la hidatidosis importada en España.

La brucelosis en cetáceos, causada por Brucella Ceti, se considera una amenaza emergente para los cetáceos, pero supone también una amenaza para la salud humana (zoonosis reemergente) y podría también serlo para las especies domésticas. La enfermedad, cuya transmisión y prevalencia no están aún determinadas, se caracteriza por afectaciones reproductivas, del sistema nervioso y en las articulaciones. La neurobrucelosis, cursando como una meningoencefalitis, es una afectación común en delfín listado (Stenella coeruleoalba). La mayoría de pruebas diagnósticas no han sido estandarizadas ni validadas para el diagnóstico de infecciones por Brucella en mamíferos marinos. Este estudio resumió y analizó la información extraída de las necropsias, la histología, la microbiología y la serología de 63 cetáceos varados en la costa catalana y de las muestras obtenidas de 12 cetáceos de la costa de la Comunidad Valenciana. Además, se realizó una caracterización patológica con una prueba imunohistoquímica con marcadores linfoides (CD3 y CD20) de los casos confirmados de neurobrucelosis. La prueba serológica con una especificidad y una sensibilidad más elevada para el diagnóstico de Brucella ceti fue el ELISA de bloqueo a una dilución 1/40 y con el punto de corte a un 50% de inhibición, evitando además el problema de las muestras hemolíticas, imposibles de analizar mediante la técnica de Rosa de Bengala (RB). La caracterización patológica de la neurobrucelosis muestra que las células implicadas en el infiltrado inflamatorio son mayoritariamente células T y células B pero con un porcentaje significativo de otras células aún por caracterizar. Este estudio refuerza la idea de que Stenella coeruleoalba (delfín listado) es una especie de cetáceo con una susceptibilidad mayor a neurobrucelosis. En referencia a la prevalencia de infección, en la situación más conservada posible, se detectó un 26,2% de animales seropositivos. Los riesgos sanitarios de esta enfermedad para los propios cetáceos, los animales domésticos y para las personas, justifican la aplicación de un programa de vigilancia para esta patología.

Freshwater mussels are known to provide ecosystem services in many ways. Their natural filter-feeding activity contributes to maintenance of water sources and also, they are considered good bio-indicators of water contamination. Unio mancus is a threatened naiad inhabiting inland freshwaters from the Mediterranean basin, whose population has been decreasing mainly due to degradation of the habitat by human activities. In the present study, U. mancus was tested as a tool for detecting Enterobacteriaceae from inland freshwater. For that purpose, two experiments were carried out. The first one evaluated the capability of these naiads to filter and retain inoculated cephalosporin-resistant E. coli in laboratory conditions. They proved to maintain bacterial loads in their soft tissue up to 4 days post-exposure. The second experiment was carried out between July 2018 and April 2019 along L'Estany de Banyoles Lake and its scraping irrigations. To determine the presence of antimicrobial-resistant Enterobacteriaceae under different levels of anthropogenic pressure, underwater caged naiads were tested for five periods of 15 consecutive days. Among all isolated bacteria using Vitek®2 system, six MDR-E. coli and one MDR-Klebsiella pneumoniae were found in the most anthroponized and polluted location points. Minimum inhibitory concentrations (MIC) for these bacteria were interpreted according to the Epidemiological cut-off (ECOFF) values from the European Committee in Antimicrobial Susceptibility Testing (EUCAST). Our results suggest that U. mancus has potential to become sentinel of bacterial pathogens of Public Health concern. A better understanding of their filtration capability will provide insights into more accurate cost-benefit analyses for their repopulation in autochthonous freshwater systems.

L’objectiu d’aquest estudi va ser evidenciar diferències, pel que fa a la càpsula fibrosa, entre els granulomes petits d’animals vacunats amb BCG (posteriorment inoculats amb M.caprae) i d’animals control (també inoculats amb M.caprae) mitjançant la tinció tricròmica de Masson, que permetessin explicar l’evolució diferent de la lesió tuberculosa en cada grup. Es van utilitzar 12 animals, 6 vacunats i 6 no vacunats (controls), en la infecció experimental, i a partir d’aquests es van obtenir els talls histològics de parènquima pulmonar, per a l’avaluació de la càpsula dels granulomes tuberculosos (o tubercles). Per a la realització d’aquesta es van tenyir els talls amb la tinció de Masson i es va avaluar qualitativament la presència de càpsula i la localització dels granulomes i semiquantitativament el gruix de la càpsula i la presència d’estroma, dividint els granulomes en dos grups en funció de la seva mida: de menys d’1 mm de diàmetre i d’1 a 5mm de diàmetre. Es va observar major grau de no encapsulament en granulomes d’animals no vacunats que en vacunats, tant en granulomes de menys d’1 mm de diàmetre com en granulomes d’1 a 5mm de diàmetre. A nivell individual es va observar que hi havia 3 dels 6 animals control que no estaven encapsulant la majoria dels granulomes d’entre 1 i 5mm de diàmetre, mentre que en el grup de vacunats només hi havia un animal poc encapsulador. Pel que fa a gruix de càpsula, es va observar un fenomen semblant, amb els animals vacunats encapsulant amb més capes els seus granulomes que els controls. Quant a la presència d’estroma, es va observar que els animals BCG en tenien més en els granulomes petits (

Introduction: Canine leishmaniosis (CanL) due to Leishmania infantum is a zoonotic protozoan disease endemic in Spain, where other vector-borne pathogens are quite common (Rickettsia conorii, Ehrlichia canis, Anaplasma spp, Hepatozoon canis, Babesia spp). Dogs are considered the main peridomestic reservoir for L. infantum infection and its clinical manifestation can vary from a total absence of clinical signs and clinicopathological abnormalities to a severe fatal clinical illness. Furthermore, there is evidence that other vector-borne organisms might affect the severity of CanL or mimic its clinical signs and/or clinicopathological abnormalities. The aim of this study was to determine co-infections with other vector-borne pathogens based on serological and molecular techniques in dogs with clinical leishmaniosis living in the Mediterranean basin and to associate them with clinical signs and clinicopathological abnormalities as well as disease severity. Materials and methods: Sixty-two dogs with clinical leishmaniosis and sixteen apparently healthy dogs were tested for Rickettsia conorii, Ehrlichia canis and Anaplasma phagocytophilum by the immunofluorescence antibody test (IFAT) and for Ehrlichia canis, Anaplasma spp, Hepatozoon spp. and Babesia spp. by polymerase chain reaction (PCR). Results: Among the dogs examined by IFAT, the seroprevalences were: 69.3% for R. conorii, 57.3% for E. canis and 44.0% for A. phagocytophilum; while the prevalences found by PCR were: 7.9% for E. canis/Anaplasma, 2.6% for Anaplasma platys and 1.3% for H. canis. Statistical association was found between dogs with clinical leishmaniosis and seroreactivity to R. conorii antigen (P = 0.025; OR = 4.09) and A. phagocytophilum antigen (P = 0.002; OR = 14.34) and being positive to more than one serological or molecular tests (co-infections) (P = 0.013) when compared with healthy dogs. Interestingly, a statistical association was found between the presence of R. conorii, E. canis and A. phagocytophilum antibodies in sick dogs and some clinicopathological abnormalities such as a decrease of albumin and albumin/globulin ratio associated to the presence of R. conorii, A. phagocytophilum and E. canis high antibody titers, and an increase in serum globulins associated to the presence of A. phagocytophilum and E. canis high antibody titers. Furthermore, seroreactivity with A. phagocytophilum antigens was statistically associated with LeishVet clinical stages III and IV. Conclusion: This study demonstrates that dogs with clinical leishmaniosis from the Barcelona and Tarragona area have a higher rate of co-infections with other vector-borne pathogens when compared with healthy controls. Furthermore, positivity to other vector-borne pathogens was associated with more pronounced clinicopathological abnormalities as well as disease severity with canine clinical leishmaniosis.

La tuberculosis (TB) es una enfermedad infecciosa que causa gran mortalidad y morbilidad globalmente en la especie humana. Mycobacterium bovis y M. caprae son las principales micobacterias patógenas para los animales, y por lo tanto las de mayor potencial zoonótico. Es necesario el desarrollo de nuevas estrategias que incrementen el control de la TB en animales, y la vacunación preventiva se considera una de las mejores opciones alternativas o complementarias a la estrategia de prueba y sacrificio. En este trabajo se evalúa la eficacia de la vacunación parenteral con una vacuna de M. bovis inactivado por calor (MBI) en cabras y sus interferencias diagnósticas. Para ello, se disponía de tres grupos de cabras (N=7 cada uno): un grupo control, un grupo vacunado con BCG, y otro grupo vacunado con MBI. Posteriormente, a todas las cabras se les inoculó M. caprae por vía endobronquial. Finalmente, los resultados de signos clínicos, las pruebas de interferón-gamma (IFN-γ) y detección de anticuerpos, la evaluación cuantitativa de lesiones tuberculosas y carga bacteriana de los tres grupos se compararon. Los resultados sugieren que la vacuna MBI protege las cabras frente a la infección, y además que la vacuna interfiere en el diagnóstico de la TB mediante la prueba del IFN-γ basada en tuberculinas, aunque esta interferencia prácticamente desaparece cuando se utilizan reactivos DIVA.

La tularemia es una enfermedad zoonótica del hemisferio norte causada por Francisella tularensis, una bacteria intracelular facultativa, que requiere una dosis infectiva muy baja y que puede causar una enfermedad potencialmente grave si no es tratada. Típicamente, las infecciones humanas y animales son causadas por cepas de la subespecie tularensis, que ocasiona brotes principalmente en América del Norte, y la holarctica, distribuida en todo el hemisferio norte, incluyendo Europa. Aunque no se conoce el reservorio natural, se ha visto que F. tularensis puede infectar una gran variedad de especies, destacando a pequeños roedores, lagormorfos y garrapatas como reservorios (y vectores) clave para esta bacteria e implicados en gran parte de los brotes reportados. Sin embargo, en Europa se ha descrito otro tipo de transmisión vectorial de la tularemia; la transmisión por mosquito, que implicaría a estos artrópodos pero también a cierto tipo de protozoos acuáticos como potenciales vectores y reservorios ambientales para F. t. holarctica. Este modo de transmisión, hasta la fecha, solo se ha documentado en regiones concretas del norte del continente europeo como Suecia o Finlandia, y no se ha podido demostrar su relevancia en otras regiones endémicas como España. Este trabajo de revisión ha consistido en hacer una recopilación de los estudios observacionales y experimentales más recientes de la literatura acerca de varios aspectos relacionados con el mantenimiento y la transmisión de F. t. holarctica en el entorno acuático, con el objetivo de poder conocer con más detalle el papel epidemiológico de los mosquitos y protozoos, así como la interacción entre ambos y el patógeno e intentar explicar el porqué de las diferencias geográficas entre diversos países en cuanto a este tipo de transmisión así como documentar las lagunas que aún siguen habiendo y proponer posibles estudios que ayuden a esclarecerlas.

[EN] Vultures are nature's most successful scavengers, feeding on the carcasses of dead animals present in the field. Availability of domestic carrion has been unstable due to rapidly changing agro-grazing economies and increasing sanitary regulations that may require burial or burning of livestock carcasses. Thus, several griffon vulture (Gyps fulvus) recoveries are based on European legislation that guarantees the animals' welfare, avoids intense persecution of the vultures and allows the feeding of threatened wildlife in supplementary feeding stations (SFS). However, in recent years, many studies have speculated on the likelihood that avian scavengers may be infected by feeding on pig carcasses at SFS from intensive livestock. In this context, the present study evaluated whether free-living griffon vultures and pig farms share zoonotic Salmonella strains to test the hypothesis that vulture are infected during consumption of carcasses provided at SFS. Here, the occurrence, serotypes and genomic DNA fingerprinting (phage typing and pulsed-field gel electrophoresis) of isolated strains were carried out in griffon vultures and pig farms authorised to provided carcasses at SFS in Castellón province (eastern Spain). The bacteriological analyses revealed that 21.1% of vultures and 14.5% for pig farms samples tested were Salmonella-positive. Monophasic S. typhimurium 1,4,[5],12:i:- was the most frequently isolated serovar. Comparison of Salmonella strains isolated from vultures and pig farms revealed that monophasic S. typhimurium 1,4,[5],12:i:-, S. Derby and S. Rissen strains were highly genetically homogeneous (similar DNA fingerprint). In conclusion, the current study indicates that free-living griffon vultures and pig farms that provide the carcasses at SFS share several zoonotic Salmonella strains. On this basis, and although transmission could be bidirectional, our result seems to corroborate the pig carcasses-to-vulture transmission and cross-infection at SFS. As an immediate Salmonella control strategy in wild avian scavengers, we suggest the implementation of a programme to guarantee that solely pig carcasses from Salmonella-free farms arrive at SFS., Funding grants from the Ministry of Infrastructures, Territory and Environment (Regional Government/Generalitat Valenciana) and from CEU-UCH (Consolidacion de Indicadores INDI15/16, INDI16/20, INDI17/25) are acknowledged. CERCA Programme from the Generalitat de Catalunya is also acknowledged.

Porcine circovirus 3 (PCV‐3) prevalence has been minimally investigated in wild boar; dynamics of infection and viral tissue distribution are currently unknown. In this study, serum samples from 518 wild boar (from years 2004 to 2018) were used to study frequency of infection. Also, serum samples from 19 boar captured and recaptured at least two times for a period of time from 1 month to 1 year were collected to determine PCV‐3 infection dynamics. Finally, to elucidate PCV‐3 DNA organic distribution, sera, different tissues and faeces were obtained from 35 additional wild boar. PCV‐3 DNA was extracted and amplified with a conventional PCR. For the PCV‐3 PCR‐positive sera from the longitudinally sampled and different tissue types, a quantitative PCR was performed. Genome sequence was obtained from a number of PCV‐3 PCR‐positive samples from different years, different time‐points of infection and tissues. Obtained results confirmed the susceptibility of wild boar to the virus, showing high frequency of PCV‐3 detection (221 out of 518, 42.66%) and demonstrating circulation at least since 2004. Compiled data indicate the possibility of long‐term infections, since 5 out of 10 PCV‐3 PCR‐positive boars longitudinally sampled showed positivity in samplings separated for more than 5 months. All tested tissue types' harboured PCV‐3 genome, with the highest percentage of PCR positivity in submandibular lymph node, tonsil, lung, liver, spleen and kidney. The amount of DNA in all tested PCV‐3 PCR‐positive samples was moderate to low. All partial and complete PCV‐3 sequences obtained from wild boar displayed high nucleotide identity, higher than 98%. In conclusion, this study further confirms that wild boar is susceptible to PCV‐3 infection, showing high frequency of detection in this animal species. Furthermore, PCV‐3 can be found in different tissues of wild boar and is apparently able to cause persistent infection. Instituto Nacional de Investigación y Tecnologia Agraria y Alimentaria. Grant Number: E‐RTA2017‐00007‐00‐00

Eastern Europe (EE) has been severely affected by mosquito-borne viruses (moboviruses). In this review, we summarize the epidemiology of moboviruses, with particular attention to West Nile virus (WNV). The study of WNV human cases in EE between 2010 and 2016, revealed that the epidemiology of WNV in EE is complex with the combination of introduction of different WNV strains from lineages 1 and 2, and the establishment of endemic cycles. We found a positive correlation between the risk of WNV re-emergence in an area and the number of human cases reported in the previous year. We also report the main ecological and biological characteristics of the key mosquito species vectors of moboviruses. Recent expansion of invasive mosquito species in EE, mainly Aedes albopictus but also Aedes aegypti and Culex quinquefasciatus, may result in new scenarios with an increased risk of transmission of moboviruses. Main gaps of knowledge in relation to moboviruses and their vectors in EE are identified. Understanding the epidemiology of moboviruses in EE is essential for the improvement of their surveillance and the control of the diseases they cause. info:eu-repo/semantics/publishedVersion

Urban wild boars (Sus scrofa) from Barcelona, Spain, harbor great diversity of Streptococcus suis strains, including strains with the cps2 gene and with the same molecular profile as local human cases. The increasing trend of potential effective contacts for S. suis transmission is of public health concern. info:eu-repo/semantics/publishedVersion

Background: Influenza A virus in swine herds represents a major problem for the swine industry and poses a constant threat for the emergence of novel pandemic viruses and the development of more effective influenza vaccines for pigs is desired. By optimizing the vector backbone and using a needle-free delivery method, we have recently demonstrated a polyvalent influenza DNA vaccine that induces a broad immune response, including both humoral and cellular immunity. Objectives: To investigate the protection of our polyvalent influenza DNA vaccine approach in a pig challenge study. Methods: By intradermal needle-free delivery to the skin, we immunized pigs with two different doses (500 μg and 800 μg) of an influenza DNA vaccine based on six genes of pandemic origin, including internally expressed matrix and nucleoprotein and externally expressed hemagglutinin and neuraminidase as previously demonstrated. Two weeks following immunization, the pigs were challenged with the 2009 pandemic H1N1 virus. Results: When challenged with 2009 pandemic H1N1, 0/5 vaccinated pigs (800 μg DNA) became infected whereas 5/5 unvaccinated control pigs were infected. The pigs vaccinated with the low dose (500 μg DNA) were only partially protected. The DNA vaccine elicited binding-, hemagglutination inhibitory (HI) − as well as crossreactive neutralizing antibody activity and neuraminidase inhibiting antibodies in the immunized pigs, in a dosedependent manner. Conclusion: The present data, together with the previously demonstrated immunogenicity of our influenza DNA vaccine, indicate that naked DNA vaccine technology provides a strong approach for the development of improved pig vaccines, applying realistic low doses of DNA and a convenient delivery method for mass vaccination. info:eu-repo/semantics/publishedVersion

Mycoplasma hyopneumoniae(M. hyopneumoniae) is the primary pathogen of enzooticpneumonia, a chronic respiratory disease in pigs. Infections occur worldwide andcause major economic losses to the pig industry. The present paper reviews the cur-rent knowledge onM. hyopneumoniaeinfections, with emphasis on identificationand analysis of knowledge gaps for optimizing control of the disease. Close contactbetween infected and susceptible pigs is the main route ofM. hyopneumoniaetrans-mission. Management and housing conditions predisposing for infection or diseaseare known, but further research is needed to better understandM. hyopneumoniaetransmission patterns in modern pig production systems, and to assess the impor-tance of the breeding population for downstream disease control. The organism isprimarily found on the mucosal surface of the trachea, bronchi and bronchioles. Dif-ferent adhesins and lipoproteins are involved in the adherence process. However, aclear picture of the virulence and pathogenicity ofM. hyopneumoniaeis still missing.The role of glycerol metabolism, myoinositol metabolism and theMycoplasmaIgbinding protein—MycoplasmaIg protease system should be further investigated fortheir contribution to virulence. The destruction of the mucociliary apparatus,together with modulating the immune response, enhances the susceptibility ofinfected pigs to secondary pathogens. Clinical signs and severity of lesions dependon different factors, such as management, environmental conditions and likely alsoM. hyopneumoniaestrain. The potential impact of strain variability on disease sever-ity is not well defined. Diagnostics could be improved by developing tests that maydetect virulent strains, by improving sampling in live animals and by designing ELI-SAs allowing discrimination between infected and vaccinated pigs. The currentlyavailable vaccines are often cost-efficient, but the ongoing research on developingnew vaccines that confer protective immunity and reduce transmission should becontinued, as well as optimization of protocols to eliminateM. hyopneumoniaefrompig herds. info:eu-repo/semantics/acceptedVersion

Bovine mastitis caused by Streptococcus uberis is a major problem in the dairy industry worldwide. Despite of that, there is no vaccine commercial available against it. Therefore, the objectives of this doctoral thesis are framed in phases 1 and 2 of the development process of a new vaccine against bovine mastitis caused by S. uberis La mamitis bovina causada per Streptococcus uberis és un problema de gran importància en les explotacions lleteres bovines a nivell mundial i en l’actualitat no existeix cap vacuna comercial que hagi demostrat ser eficaç per combatre aquesta patologia. Els objectius de la present tesi doctoral s’emmarquen en les fases 1 i 2 del procés de desenvolupament d’una nova vacuna contra la mamitis bovina causada per S. uberis

Las enfermedades transmitidas por garrapatas a humanos han mostrado un aumento en los reportes en los últimos años en Europa. Se considera que factores medioambientales y antropológicos serían los responsables. No obstante, hay que tener en cuenta que cada vez hay más interés investigativo en el tema, los sistemas de vigilancia de varios países europeos han incluido a algunas de estas enfermedades bajo sus listas de notificación y ha mejorado la comunicación hacia profesionales de la salud y la población en general. Esta revisión describe el estado epidemiológico de Fiebre hemorrágica Crimea-Congo, Encefalitis por garrapatas, Fiebre botonosa del Mediterráneo, Fiebre recurrente por garrapatas y Enfermedad de Lyme, según la información publicada en fuentes oficiales y artículos científicos del nuevo milenio. Sin embargo, debido a que la disponibilidad de datos no es satisfactoria en todos los casos, es probable que de manera general el riesgo esté subestimado en Europa. En contraste con esto, un enfoque interdisciplinar en la vigilancia e investigación es necesario para que las brechas de información puedan cada vez ser más estrechas y la prevención de estas enfermedades sea eficiente y oportuna tanto para los animales como para los humanos. Abstract Tick-borne diseases in humans have shown an increase in their reports in the last years. Several environmental and anthropological factors have been considered as responsible, but it is also important to say that while time passes there is more interest about this topic in scientific community; lots of countries have included these diseases as notifiable and there has been improvement in communication towards health professionals and general population. This review describes Crimean-Congo hemorrhagic fever, tick-borne encephalitis, Mediterranean spotted fever, tick-borne relapsing fever and Lyme disease epidemiological status according to the information available in official sources and scientific articles, however, due to lack of information in some cases it's possible that, in general, the risk is underestimated. According to this, a one health approach in surveillance and research is needed so that information gaps that exist today could be reduced in the future, as a result, prevention would be more efficient and opportune for animal and human health.

El brote que asoló Guinea, Liberia y Sierra Leona entre 2013 y 2016 representa un cambio en la presentación de esta enfermedad, dándose por primera vez en áreas urbanas y con movimiento de personas. Esto junto con las pobres condiciones del sistema sanitario de los tres países afectados permitió que el brote escalara rápidamente y a posterior que se dieran casos hasta en 7 países diferentes con transmisión nosocomial. Este brote ha dejado patente que existe un riesgo, aunque pequeño, de importación de casos al primer mundo demostrando aquí que el Ébola no es solo un problema de África sino un problema a escala mundial con consecuencias devastadoras. Para saber cómo se gestionó la alerta sanitaria durante ese brote se ha hecho una búsqueda bibliográfica que incluían las palabras "Ebola" o "Ebolavirus" solas o acompañadas por "disease", "perspective" o "present" o la combinación de palabras "West Africa Outbreak".

Nowadays, antibiotic resistance is one of the main public health problems. Finding alternative strategies to the use of antibiotics in veterinary medicine is one of the great challenges. In poultry production (and in other food animals) some of these possible alternatives would be: a) to take advantage of bacterial cell to cell communication (quorum sensing) and the disruption of that communication (quorum quenching) to control the proliferation pathogens; b) the use of additives in animal feed which modulate the intestinal microbiota, promoting bacterial populations with antibacterial activity. In this study we aimed to: a) to investigate the production of acylhomoserine lactone (AHL) by Yersinia enterocolitica 057 under different conditions and its presence in the gut contents of chickens; b) to investigate the potential antimicrobial activity of the intestinal microbiota against different pathogens in three groups of chickens fed with different feed. These different diets (conventional diet, diet with xylanase, diet with xylanase and xylooligosaccharides) may promote different intestinal microbiota, and thus different antimicrobial activity, if any. The results suggest that the intestinal microbiota may interfere in the growth of Y. enterocolitica and/or its production of AHL. On the other hand, no antimicrobial activity against the pathogens tested could be detected in any of the 3 groups of chickens. The different growth conditions may have influenced the different bacterial populations able to grow on the agar plates and therefore the bacteria with potential antimicrobial activity.

La salud humana va ligada a relaciones intra-especie, inter-especie y al entorno. Dentro de los riesgos asociados a dichas relaciones inter-especie se encuentran las zoonosis transmitidas por mascotas. Es por ello que todo cuanto los propietarios y sus mascotas hagan, influirá tanto en el bienestar humano como en el medio, ya que en una sociedad la salud de todos sus miembros se encuentra interconectada. Para reducir el impacto de los factores de riesgo por parte de los propietarios en relación a sus mascotas, estos deben recibir toda la información necesaria con el fin de reducir los peligros asociados (políticas de prevención y control de patógenos). Se realizó un cuestionario a propietarios de perros y gatos en dos centros veterinarios de la provincia de Barcelona, con el fin de averiguar el grado de conocimiento que estos tienen sobre zoonosis transmitidas por mascotas. De un total de 150 encuestas, se realizaron 120 en el Hospital Clínic Veterinari de la Universitat Autònoma de Barcelona y el resto un consultorio veterinario en el municipio de Santa Coloma de Gramenet. Con los resultados obtenidos se realizaron tablas de contingencia, relaciones entre variables, cálculo de chi cuadrado y gráficos de interpretación. Se observan resultados remarcables, considerados factores de riesgo, tales como el 71% de los encuestados permite un contacto más o menos frecuente de su rostro con el hocico de la mascota y un 89% nunca ha sido informado sobre zoonosis transmitidas por mascotas. También se analizó la información relacionada a este tema mediante búsqueda bibliográfica, datos de uso público y mediante dos entrevistas (a un doctor en veterinaria y uno en medicina). Tras el cuestionario realizado y el análisis de información, se concluyó que se necesita reforzar la comunicación sobre zoonosis transmitidas por mascotas a los propietarios. Dicha tarea es una labor conjunta de veterinarios, médicos y Salud Pública (Una Salud, Una Comunicación).

Background: Low levels of vitamin D are associated with cardiovascular diseases in adults; however, it is unclear if there is a relationship between vitamin D in pregnancy and cardiovascular diseases in offspring. Aim: To assess the association of maternal circulating 25-hydroxyvitamin D (25(OH)D) at 12 weeks of pregnancy and child concentrations of (25(OH)D) in preschool age with offspring pulse wave velocity, retinal microcirculation and blood pressure at 11 years. Methods: We included 401 mother-child pairs from the INMA-Sabadell cohort study. We used plasma (25(OH)D) at first trimester of pregnancy and offspring plasma at 4 years. Main outcomes measured at 11 years were pulse wave velocity, central venular retina equivalent (CRVE), central retinal arteriolar equivalent (CRAE), and systolic and diastolic blood pressures. We used (25(OH)D) as a continuous and categorical variable: deficiency (≤20 ng/ml), insufficiency (21-30 ng/ml), and sufficiency (>30 ng/ml). Results: The median plasma levels of 25(OH)D3 were 29.63 and 37.17 ng/ml in pregnancy and childhood, respectively. Higher maternal vitamin D levels were associated with an increase in systolic blood pressure [beta = 1.06 (95% CI=0.14, 1.97)] but not with diastolic blood pressure. Maternal vitamin D levels were not associated with pulse wave velocity or with microcirculation. Vitamin D in prescholar age was not associated with any of the cardiovascular outcomes assessed. Conclusions: This study finds little evidence of associations between vitamin D levels during pregnancy or in pre-school age with cardiovascular outcomes in children. Further studies will be needed to understand more clearly the influence of prenatal and early life vitamin D levels on cardiovascular disease in childhood.

The objective of this study was to investigate the association between teat skin colonization and intramammary infection (IMI) with Staphylococcus aureus or Streptococcus agalactiae at the quarter level in herds with automatic milking systems. Milk and teat skin samples from 1,142 quarters were collected from 300 cows with somatic cell count >200,000 cells/mL from 8 herds positive for Strep. agalactiae. All milk and teat skin samples were cultured on calf blood agar and selective media. A subset of samples from 287 quarters was further analyzed using a PCR assay (Mastit4 PCR; DNA Diagnostic A/S, Risskov, Denmark). Bacterial culture detected Staph. aureus in 93 (8.1%) of the milk samples and 75 (6.6%) of the teat skin samples. Of these, 15 (1.3%) quarters were positive in both the teat skin and milk samples. Streptococcus agalactiae was cultured in 84 (7.4%) of the milk samples and 4 (0.35%) of the teat skin samples. Of these, 3 (0.26%) quarters were positive in both the teat skin and milk samples. The PCR detected Staph. aureus in 29 (10%) of the milk samples and 45 (16%) of the teat skin samples. Of these, 2 (0.7%) quarters were positive in both the teat skin and milk samples. Streptococcus agalactiae was detected in 40 (14%) of the milk samples and 51 (18%) of the teat skin samples. Of these, 16 (5.6%) quarters were positive in both the teat skin and milk samples. Logistic regression was used to investigate the association between teat skin colonization and IMI at the quarter level. Based on bacterial culture results, teat skin colonization with Staph. aureus resulted in 7.8 (95% confidence interval: 2.9; 20.6) times higher odds of Staph. aureus IMI, whereas herd was observed as a major confounder. However, results from the PCR analyses did not support this association. Streptococcus agalactiae was isolated from the teat skin with both PCR and bacterial culture, but the number of positive teat skin samples detected by culture was too low to proceed with further analysis. Based on the PCR results, Strep. agalactiae on teat skin resulted in 3.8 (1.4; 10.1) times higher odds of Strep. agalactiae IMI. Our results suggest that Staph. aureus and Strep. agalactiae on teat skin may be a risk factor for IMI with the same pathogens. Focus on proper teat skin hygiene is therefore recommended also in AMS. info:eu-repo/semantics/publishedVersion

La Leishmaniosis visceral humana causada por Leishmania donovani es una enfermedad antroponótica endémica en muchos países de África oriental. Sin embargo, se desconoce si animales domésticos o salvajes pueden tener un papel en esta infección. El principal objetivo de este estudio fue detectar anticuerpos frente a Leishmania infantum en sueros de perros del noroeste de Uganda mediante ELISA indirecto y evaluar los factores de riesgo. Para ello, se analizaron muestras de suero del año 2017 y muestras de suero impregnadas en papel de filtro Whatman nº3 del 2018. El trabajo de campo se llevó a cabo en los Parques Nacionales de Elizabeth Queen (QENP), Mgahinga, Budongo y en los distritos de Kasese, Kapchorwa, Buliisa, Masindi, Kibingo, Hoima y Kanungu. Las muestras de suero del 2017 fueron todas negativas. La seroprevalencia global obtenida de las muestras impregnadas en papel filtro fue del 50.8%, siendo los distritos de Karusandara y Kilembe los que obtuvieron una mayor proporción de perros seropositivos. Entre los datos de frecuencia analizados, no se encontró ninguna asociación estadísticamente significativa entre las variables estudiadas. Si bien, se observaron valores estadísticamente significativos con respecto a porcentajes de positividad registrados por localidad y condición corporal de los perros muestreados. Los resultados del presente estudio sugieren el posible carácter endémico de la leishmaniosis canina en el noreste de Uganda. No obstante, estudios futuros son necesarios para determinar la prevalencia real de esta infección y su correación con otras tripanosomiasis en todo el país y evaluar el verdadero rol de los perros como reservorios a la leishmaniosis en esta área geográfica.

Altres ajuts: Secretaria del Departament d'Economia i Creixement de la Generalitat de Catalunya DI2013-0039 Information on the in vitro growth of pathogenic and non-pathogenic Mycoplasma hyopneumoniae (M. hyopneumoniae) strains is scarce and controversial. Despite its limitations, the colour changing units (CCU) assay is still considered the golden standard titration technique for M. hyopneumoniae culture. Thus, the aims of the present study were: (1) to describe the growth dynamics and kinetics of pathogenic and non-pathogenic M. hyopneumoniae strains, and (2) to monitor the strains' daily growth by ATP luminometry, CCU, colony forming units (CFU), and DNA quantification by real time quantitative PCR (qPCR) and by fluorescent double-stranded DNA (dsDNA) staining, to evaluate them as putative titration methodologies. The growth of the non-pathogenic J (ATCC ® 25934™) type strain and the pathogenic 11 (ATCC ® 25095™) reference strain and 232 strain was modelled by the Gompertz model. Globally, all three-strain cultures showed the same growing phases as well as similar maximal titres within a particular technique, but for CFU. However, the J strain displayed the fastest growing. During the logarithmic phase of growing, CCU, ATP and M. hyopneumoniae copy titres were strongly and linearly associated, and correlation between techniques could be reliably established. In conclusion, real-time culture titration by means of ATP or molecular assays was useful to describe the in vitro growth of the tested strains. Knowledge about the in vitro growth behaviour of a specific strain in a specific medium may provide several advantages, including information about the time required to reach maximal titres by the culture. Noteworthy, the obtained results refers to the three strains used, so extrapolation to other M. hyopneumoniae strains or culture conditions should be made cautiously.

Dengue has emerged as the most important viral mosquito-borne disease globally. The current risk of dengue outbreaks in Europe appeared with the introduction of the vector Aedes albopictus mosquito in Mediterranean countries. Considering the increasing frequency of dengue epidemics worldwide and the movement of viraemic hosts, it is expected that new autochthonous cases will occur in the future in Europe. Arbovirus surveillance started in Catalonia in 2015 to monitor imported cases and detect possible local arboviral transmission. During 2015, 131 patients with a recent travel history to endemic countries were tested for dengue virus (DENV) and 65 dengue cases were detected. Twenty-eight patients with a febrile illness were viraemic, as demonstrated by a positive real-time RT-PCR test for DENV in serum samples. Entomological investigations around the viraemic cases led to the detection of DENV in a pool of local Ae. albopictus captured in the residency of one case. The sequence of the DENV envelope gene detected in the mosquito pool was identical to that detected in the patient. Our results show how entomological surveillance conducted around viraemic travellers can be effective for early detection of DENV in mosquitoes and thus might help to prevent possible autochthonous transmission. Funding acknowledgment to the CERCA Programme/Generalitat de Catalunya and the Health department of the Generalitat de Catalunya. The study was also partially funded by ISCIII, project RD12/0018/0006 Red de Enfermedades Tropicales, Subprograma RETICS del Plan Estatal de I+D+I 2013-2016 and cofunded by FEDER: Una manera de hacer Europa and by grant 2017 SGR 794 from the Agència de gestió Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR) - Departament d’Empresa i Coneixement . Generalitat de Catalunya. The authors are very grateful for the excellent technical contribution of Núria Pujol and Raquel Rivas from CReSA (IRTA). Sí

Dromedary camel heavy chain–only antibodies may provide novel intervention strategies against MERS coronavirus., Middle East respiratory syndrome coronavirus (MERS-CoV) continues to cause outbreaks in humans as a result of spillover events from dromedaries. In contrast to humans, MERS-CoV–exposed dromedaries develop only very mild infections and exceptionally potent virus-neutralizing antibody responses. These strong antibody responses may be caused by affinity maturation as a result of repeated exposure to the virus or by the fact that dromedaries—apart from conventional antibodies—have relatively unique, heavy chain–only antibodies (HCAbs). These HCAbs are devoid of light chains and have long complementarity-determining regions with unique epitope binding properties, allowing them to recognize and bind with high affinity to epitopes not recognized by conventional antibodies. Through direct cloning and expression of the variable heavy chains (VHHs) of HCAbs from the bone marrow of MERS-CoV–infected dromedaries, we identified several MERS-CoV–specific VHHs or nanobodies. In vitro, these VHHs efficiently blocked virus entry at picomolar concentrations. The selected VHHs bind with exceptionally high affinity to the receptor binding domain of the viral spike protein. Furthermore, camel/human chimeric HCAbs—composed of the camel VHH linked to a human Fc domain lacking the CH1 exon—had an extended half-life in the serum and protected mice against a lethal MERS-CoV challenge. HCAbs represent a promising alternative strategy to develop novel interventions not only for MERS-CoV but also for other emerging pathogens.

Porcine circovirus 2 (PCV-2) is a virus characterized by a high evolutionary rate, promoting the potential emergence of diferent genotypes and strains. Despite the likely relevance in the emergence of new PCV-2 variants, the subtle evolutionary patterns of PCV-2 at the individual-host level or over short transmission chains are still largely unknown. This study aimed to analyze the within-host genetic variability of PCV-2 subpopulations to unravel the forces driving PCV-2 evolution. A longitudinal weekly sampling was conducted on individual animals located in three farms after the frst PCV-2 detection. The analysis of polymorphisms evaluated throughout the full PCV-2 genome demonstrated the presence of several single nucleotide polymorphisms (SNPs) especially in the genome region encoding for the capsid gene. The global haplotype reconstruction allowed inferring the virus transmission network over time, suggesting a relevant within-farm circulation. Evidences of co-infection and recombination involving multiple PCV-2 genotypes were found after mixing with pigs originating from other sources. The present study demonstrates the remarkable within-host genetic variability of PCV-2 quasispecies, suggesting the role of the natural selection induced by the host immune response in driving PCV-2 evolution. Moreover, the efect of pig management in multiple genotype coinfections occurrence and recombination likelihood was demonstrated. info:eu-repo/semantics/publishedVersion

Middle East respiratory syndrome (MERS) represents an important respiratory disease accompanied by lethal outcome in one-third of human patients. Recent data indicate that dromedaries represent an important source of infection, although information regarding viral cell tropism and pathogenesis is sparse. In the current study, tissues of eight dromedaries receiving inoculation of MERS-Coronavirus (MERS-CoV) after recombinant Modified-Vaccinia-Virus-Ankara (MVA-S)-vaccination (n = 4), MVA-vaccination (mock vaccination, n = 2) and PBS application (mock vaccination, n = 2), respectively, were investigated. Tissues were analyzed by histology, immunohistochemistry, immunofluorescence, and scanning electron microscopy. MERS-CoV infection in mock-vaccinated dromedaries revealed high numbers of MERS-CoV-nucleocapsid positive cells, T cells, and macrophages within nasal turbinates and trachea at day four post infection. Double immunolabeling demonstrated cytokeratin (CK) 18 expressing epithelial cells to be the prevailing target cell of MERS-CoV, while CK5/6 and CK14 expressing cells did not co-localize with virus. In addition, virus was occasionally detected in macrophages. The acute disease was further accompanied by ciliary loss along with a lack of dipeptidyl peptidase 4 (DPP4), known to mediate virus entry. DPP4 was mainly expressed by human lymphocytes and dromedary monocytes, but overall the expression level was lower in dromedaries. The present study underlines significant species-specific manifestations of MERS and highlights ciliary loss as an important finding in dromedaries. The obtained results promote a better understanding of coronavirus infections, which pose major health challenges. info:eu-repo/semantics/publishedVersion

Mycobacterium bovis (M.bovis) is the main causative agent for bovine tuberculosis (BTB) and can also be the cause of zoonotic tuberculosis in humans. In view of its zoonotic nature, slaughterhouse surveillance, potentially resulting in total or partial condemnation of the carcasses and organs, is conducted routinely. Spoligotyping, VNTR profiling, and whole genome sequencing (WGS) of M. bovis isolated from tissues with tuberculosis-like lesions collected from 14 cattle at Eritrea’s largest slaughterhouse in the capital Asmara, were conducted.The 14 M. bovis isolates were classified into three different spoligotype patterns (SB0120, SB0134 and SB0948) and six VNTR profiles. WGS results matched those of the conventional genotyping methods and further discriminated the six VNTR profiles into 14 strains. Furthermore, phylogenetic analysis of the M. bovis isolates suggests two independent introductions of BTB into Eritrea possibly evolving from a common ancestral strain in Europe.This molecular study revealed the most important strains of M. bovis in Eritrea and their (dis)similarities with the strains generally present in East Africa and Europe, as well as potential routes of introduction of M. bovis. Though the sample size is small, the current study provides important information as well as platform for future in-depth molecular studies on isolates from both the dairy and the traditional livestock sectors in Eritrea and the region. This study provides information onthe origin of some of the M. bovis strains in Eritrea, its genetic diversity, evolution and patterns of spread between dairy herds. Such information is essential in the development and implementation of future BTB control strategy for Eritrea., Author summary The livestock sector plays a major role in poverty and hunger reduction in the vast majority of Africa, as a source of food, cash income, manure, draught power, transportation, savings, insurance and social status. However, for livestock to play this vital role, the impact of diseases of economic and zoonotic importance need to be reduced. Bovine tuberculosis, mainly caused by Mycobacterium bovis, is such an infectious disease. Slaughterhouse (gross pathology) surveillance, followed by bacterial culture and genotyping, are options to identify the disease-causing agents, their distribution, and enabling trace back of the sources of infections, in order to prevent their re-introduction and spread. Unfortunately, genotyping is by far not generally introduced in the continent. In the present study, tissues with tuberculosis-like lesions were collected from the Asmara municipal slaughterhouse, the largest slaughterhouse in Eritrea, and bacterial culture, classical Mycobacterium tuberculosis complex typing (Spoligotyping and VNTR profiling), as well as whole genome sequencing (WGS) were used to gain insight into the spatial and temporal distribution, genetic diversity and evolution of M. bovis strains circulating in Eritrean dairy cattle. The results revealed (dis)similarities of the Eritrean M. bovis strains with the strains generally present in Africa and Europe, potential routes of introduction to Eritrea and genetic diversity of the M. bovis strains. Future in-depth molecular studies including more samples from dairy cattle as well as cattle and goats from the traditional livestock sector are recommended.

Background This study sought to evaluate the effect of sow vaccination against Porcine circovirus 2 (PCV2) on reproductive parameters during two consecutive reproductive cycles. The study was performed in a PCV2 subclinical infected breeding herd (PCV2 circulation but absence of major reproductive problems). Ninety-four pregnant sows were primo-immunized with a commercial PCV2 vaccine and ninety-seven were injected with phosphate-buffered saline at 6 and 3 weeks before the first studied farrowing, and then boosted at 2 weeks before the second one. Blood samples were taken throughout the study to assess PCV2 DNA load and antibodies. At farrowing, main reproductive parameters and piglet vitality index were registered. In addition, in those litters with more than three mummified or stillborn piglets, microscopic examination and PCV2 antigen detection in foetal myocardium was done. Results Vaccinated sows showed significantly higher antibody levels compared to the non-vaccinated counterparts. PCV2 DNA was only detected at farrowing in 2 (4.2%) non-vaccinated sows. Vaccinated sows had 1.3 more live-born piglets per litter at the second cycle than non-vaccinated counterparts. Piglets from vaccinated sows had significantly higher (+ 12.7%) vitality score than the ones born from non-vaccinated sows. No PCV2 compatible lesions neither PCV2 antigen were detected in the tested foetal hearts. Conclusions The present study represents a first attempt to demonstrate that PCV2 sow vaccination may have a positive influence on prolificacy and vitality of the offspring in a subclinical infected breeding herd. However, since reproductive outcomes at farm level may be affected by a number of factors, further studies would be needed to confirm this association.

Foot-and-mouth disease (FMD) is a highly contagious viral disease affecting cloven-hoofed animals that causes severe economic losses. The disease is characterized by a vesicular condition and it cannot be differentiated from other vesicular diseases. Therefore, laboratory confirmation of any suspected FMD case is compulsory. Despite viral isolation in cell cultures has been considered for many years as the gold standard for FMD diagnosis, the advantages of real-time reverse transcription polymerase chain reaction (rRT-PCR) technology have motivated its use directly in clinical specimens for FMD diagnosis. The current work was aimed to develop and validate a molecular multi-check strategy using rRT-PCR (mMulti-rRT-PCR) based on SYBR-Green I for pan/foot-and-mouth disease virus (pan/FMDV) diagnosis. From in silico approaches, different primer pairs previously reported were selected and modified to reduce the likelihood of viral escape as well as potential failures in the pan/FMDV detection. The analytical parameters were evaluated using a high number of representative viral strains. The repeatability of the assay and its performance on field samples were also assessed. The mMulti-rRT-PCR was able to detect emergent FMDV strains that circulated in South America between the years 2006–2010 and on which the single rRT-PCRs failed when they were applied independently. The results obtained here showed that the proposed system is an accurate and rapid diagnosis method for sensitive and specific detection of FMDV. Thus, a validated mMulti-rRT-PCR assay based on SYBR-Green I detection coupled to melting curves resolution for pan/FMDV diagnosis on clinical samples is proposed. This study also highlights the need to incorporate the multi-target detection principle in the diagnosis of highly variable agents, specially, of those listed by OIE like FMDV. info:eu-repo/semantics/publishedVersion

Background Neonatal diarrhoea represents a major disease problem in the early stages of animal production, increasing significantly pre-weaning mortality and piglets weaned below the target weight. Enteric diseases in newborn piglets are often of endemic presentation, but may also occur as outbreaks with high morbidity and mortality. The objective of this study was to assess the frequency of different pathogens involved in cases of recurrent neonatal diarrhoea in Spain. Results A total of 327 litters from 109 sow farms located in Spain with neonatal recurrent diarrhoea were sampled to establish a differential diagnosis against the main enteric pathogens in piglets. In total, 105 out of 109 (96.3%) case submissions were positive to one of the examined enteric organisms considered potentially pathogenic (Escherichia coli, Clostridium perfringens types A and C, Transmissible gastroenteritis virus [TGEV], Porcine epidemic diarrhoea virus [PEDV] or Rotavirus A [RVA]). Fifty-eight out of 109 (53.2%) submissions were positive for only one of these pathogens, 47 out of 109 (43.1%) were positive for more than one pathogen and, finally, 4 out of 109 (3.7%) were negative for all these agents. Escherichia coli strains were isolated from all submissions tested, but only 11 of them were classified into defined pathotypes. Clostridium perfringens type A was detected in 98 submissions (89.9%) and no C. perfringens type C was found. Regarding viruses, 47 (43.1%) submissions were positive for RVA, 4 (3.7%) for PEDV and none of them for TGEV. Conclusion In conclusion, C. perfringens type A, E. coli and RVA were the main pathogens found in faeces of neonatal diarrheic piglets in Spain.

Porcine reproductive and respiratory syndrome virus (PRRSv) infection alters the host’s cellular and humoral immune response. Immunity against PRRSv is multigenic and vary between individuals. The aim of the present study was to compare several genes that encode for molecules involved in the immune response between two groups of vacci‑ nated pigs that experienced short or long viremic periods after PRRSv challenge. These analyses include the sequenc‑ ing of four SLA Class I, two Class II allele groups, and CD163, plus the analysis by quantitative realtime qRT-PCR of the constitutive expression of TLR2, TLR3, TLR4, TLR7, TLR8 and TLR9 mRNA and other molecules in peripheral blood mononuclear cells. info:eu-repo/semantics/publishedVersion

Tuberculosis (TB) in animals is a re-emerging disease with a wide range of hosts that causes large economic losses in livestock. Goats are particularly susceptible to TB and, in endemic areas, vaccination may be a valuable measure to control the disease. The main aim of this study was to evaluate the efficacy of parenteral vaccination of goats with a heatinactivated Mycobacterium bovis (HIMB) vaccine, and compare it to M. bovis Bacille Calmette–Gue ´rin (BCG) vaccine. Twenty-four goat kids were divided in 3 groups as following: HIMB vaccinated group (n = 8), BCG vaccinated group (n = 8) and unvaccinated group (n = 8). Afterwards, goats were experimentally challenged with Mycobacterium caprae by the endobronchial route. Antigen specific interferon-γ release assays and serology were performed after vaccination and challenge. Pathological and bacteriological parameters were evaluated after necropsy at 9 weeks post-challenge (p.c.). HIMB vaccine showed similar levels of protection to BCG in terms of volume reduction of thoracic TB lesions, presence of extra-pulmonary lesions, as well as a slight reduction of bacterial load in pulmonary lymph nodes. Moreover, HIMB vaccine did not induce interferences on the interferon-γ release assay based on reagents previously developed to differentiate infected from BCG vaccinated individuals. The results indicate that HIMB is a suitable vaccine candidate for further larger-scale trials under field conditions in goats. info:eu-repo/semantics/publishedVersion

Background: Bovine cysticercosis (BCC) (due to Taenia saginata) is often claimed to cause considerable economic losses to the livestock industry, particularly in beef cattle, but recent studies estimating the economic impact are lacking. The current study aimed to quantify the annual economic impact of BCC in Belgium from 2012 to 2016, by gathering data from diverse sources in the meat and human health sectors. Results: In Belgium, on average, 15 carcasses with generalised infections and 1168 carcasses with localised ones are detected upon meat inspection each year. The highest proportion of the total economic losses due to bovine cysticercosis were borne by the cattle owners with an average economic cost of €3,408,455/year: €2,954,061/year due to BCC insurance, €453,024/year due to value losses of beef of uninsured carcasses (i.e. freezing process) and €1370/year due to destruction costs of uninsured carcasses with generalised infections. The slaughterhouses suffered an economic impact of €210,806/year. They were responsible for inspection costs related to meat inspection in general, administration, processing and deboning of infected carcasses (€597,856/year), value losses (€34,848/year) and destruction costs (€105/year) of carcasses insured by the slaughterhouses (unofficial insurance) (5% of slaughtered animals). On the other hand, the slaughterhouses gained a total of €422,004/year due to unofficial insurance fees. Thirty percent of all slaughtered animals were officially insured against BCC and the insurance company generated an income of €2,322,337/year. The economic impact related to taeniosis (10,991 patients annually) amounted to a maximum of €795,858/year. Conclusion: BCC and taeniosis due to T. saginata have a large economic impact in Belgium, mainly due to the insurance costs for BCC. These results indicate the need for reducing the number of BCC and taeniosis cases to avoid the costs and losses related to this parasite. Keywords: Taenia saginata, Bovine cysticercosis, Economic impact info:eu-repo/semantics/publishedVersion

Since Schmallenberg disease was discovered in 2011, the disease rapidly spread across Europe. Culicoides biting midges have been implicated as putative Schmallenberg vectors in Europe. The detection of Schmallenberg virus (SBV) in field collected Culicoides was evaluated through retrospective (2011–2012) collections and captures performed in 2013. This study represents the first detection of SBV in field collected Culicoides in Spain. Infectious midges were detected at the foothills of Pyrenees, Aramunt, in the summer 2012. All the specimens infected with Schmallenberg were of the species Culicoides obsoletus s.s. confirming its putative vector status in Spain. Experimental infection on field collected Culicoides provided evidence of atypical high efficiency for SBV vector infection and transmission potential in local populations of Culicoides imicola and in Culicoides of the Obsoletus complex. However, captured individuals of C. imicola were more susceptible to SBV infection than C. obsoletus s.l. (p < .001), with an infection ratio of 0.94 and 0.63, respectively. In contrast, a Culicoides nubeculosus colony appeared to be refractory to SBV infection. info:eu-repo/semantics/acceptedVersion

The family Asfarviridae includes the single species African swine fever virus, isolates of which have linear dsDNA genomes of 170–194 kbp. Virions have an internal core, an internal lipid membrane, an icosahedral capsid and an outer lipid envelope. Infection of domestic pigs and wild boar results in an acute haemorrhagic fever with transmission by contact or ingestion, or by ticks of the genus Ornithodoros. Indigenous pigs act as reservoirs in Africa, where infection is endemic, and from where introductions occur periodically to Europe. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the taxonomy of the Asfarviridae, which is available at www.ictv.global/report/asfarviridae. info:eu-repo/semantics/publishedVersion

Indexación: Scopus. The increase in human population and domestic pets, such as cats, are generating important consequences in terms of habitat loss and pathogen pollution of coastal ecosystems with potential to generate negative impacts in marine biodiversity. Toxoplasma gondii is the etiological agent of zoonotic disease toxoplasmosis, and is associated with cat abundance and anthropogenic disturbance. The presence of T. gondii oocysts in the ocean has negatively affected the health status of the threatened Southern sea otter (Enhydra lutris nereis) populations. The present study analyzed seroprevalence and presence of T. gondii DNA in American mink (Neovison vison), Southern river otters (Lontra provocax) and domestic cats (Felis silvestris catus) in four different areas in Southern Chile comprising studies in rivers and lakes in Andean foothills and mountains, marine habitat and island coastal ecosystems. Mean seroprevalence of T. gondii in the study was 64% of 151 total animals sampled: 59% of 73 American mink, 77% of 13 Southern river otters, 68% of 65 domestic cats and in two of two kodkods (Leopardus guigna). Toxoplasma gondii DNA was detected in tissues from one American mink and one Southern river otter. The present study confirms the widespread distribution of T. gondii in Southern Chile, and shows a high exposure of semiaquatic mustelids and domestic cats to the parasite. Cats and anthropogenic disturbance have a role in the maintenance of T. gondii infection in ecosystems of southern Chile. © 2018 Barros et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0199085

Medical imaging has greatly progressed to become an essential technology in current clinical processes. However, advances in diagnostic imaging have not been applied to the same extent to other fields such as education and meat science. In this thesis two e-learning platforms are presented to improve teaching methodologies in medical and veterinary teaching, focusing mainly in the ease for the content creation and the image interaction, and supporting multiple graphical resources such as 3D models. An image processing algorithm is also presented to improve the quality classification process of farm animals by computing the lean meat percentage, either from carcass or live animals images, proposing for the latter two segmentation algorithms to remove the internal organs La imatge mèdica ha progressat a bastament per convertir-se en una tecnologia imprescindible en els processos clínics actuals. Tanmateix, els avenços en la imatge per al diagnòstic no s'han aprofitat de la mateixa manera en altres camps com ara l'educació o la ciència de la carn. En aquesta tesi es presenten dues plataformes d'aprenentatge en línia per a millorar els processos d'aprenentatge en la docència mèdica i veterinària, centrant-se sobretot en la facilitat per a la creació de contingut i en la interacció amb la imatge, i donant suport a múltiples recursos gràfics com ara models 3D. També es presenta un algorisme de processament d'imatge per millorar el procés de classificació de la qualitat d'animals de producció calculant el percentatge de carn magra, ja sigui a partir d'imatges de canals o d'animals vius, proposant dos algorismes de segmentació per eliminar els òrgans interns en aquest últim cas

Very solid evidence suggests that the core of full length PrPSc is a 4-rung β-solenoid, and that individual PrPSc subunits stack to form amyloid fibers. We recently used limited proteolysis to map the β-strands and connecting loops that make up the PrPSc solenoid. Using high resolution SDS-PAGE followed by epitope analysis, and mass spectrometry, we identified positions ~116/118, 133–134, 141, 152–153, 162, 169 and 179 (murine numbering) as Proteinase K (PK) cleavage sites in PrPSc. Such sites likely define loops and/or borders of β-strands, helping us to predict the threading of the β-solenoid. We have now extended this approach to recombinant PrPSc (recPrPSc). The term recPrPSc refers to bona fide recombinant prions prepared by PMCA, exhibiting infectivity with attack rates of ~100%. Limited proteolysis of mouse and bank vole recPrPSc species yielded N-terminally truncated PK-resistant fragments similar to those seen in brain-derived PrPSc, albeit with varying relative yields. Along with these fragments, doubly N- and C-terminally truncated fragments, in particular ~89/97-152, were detected in some recPrPSc preparations; similar fragments are characteristic of atypical strains of brain-derived PrPSc. Our results suggest a shared architecture of recPrPSc and brain PrPSc prions. The observed differences, in particular the distinct yields of specific PK-resistant fragments, are likely due to differences in threading which result in the specific biochemical characteristics of recPrPSc. Furthermore, recombinant PrPSc offers exciting opportunities for structural studies unachievable with brain-derived PrPSc., Author summary PrPSc Prions propagate by inducing the refolding of the natively folded normal cellular prion protein (PrPC) into the prion conformation in brain and other mammalian tissues (wild-type). Understanding the structure of PrPSc is essential to understanding how PrPSc prions propagate. The secondary structure of PrPC is composed of four α-helical regions, two very small β-sheets and random coil, while PrPSc is composed entirely of β-sheets and random coil. The β-sheets of PrPSc wind four times to form a spring-like 4-rung β-solenoid. The rungs are connected by stretches of random coil. We have used proteinase K (PK), to cleave these random coil connecters, allowing us to identify the location of the more PK-resistant β-strand stretches within wild-type PrPSc. In this work, we use recombinant PrPSc, (in vitro generated infectious prions) to show that patterns of PK cleavage for recombinant and wild-type PrPSc are very similar, indicating that they share a common architecture. This also means that recombinant PrPSc is a true surrogate for wild-type PrPSc. Since recombinant PrPSc is derived from recombinant PrP, future structural studies employing specific amino acid or stable-isotope labeled amino acid substitutions are easily achievable. Such substitutions are essential for NMR studies.

International audience; Background The lumpy skin disease virus (LSDV) is a dsDNA virus belonging to the Poxviridae family and the Capripoxvirus genus. Lumpy skin diseases (LSD) is a highly contagious transboundary disease in cattle producing major economic losses. In 2014, the disease was first reported in the European Union (in Cyprus); it was then reported in 2015 (in Greece) and has spread through different Balkan countries in 2016. Indirect vector transmission is predominant at small distances, but transmission between distant herds and between countries usually occurs through movements of infected cattle or through vectors found mainly in animal trucks. Methods and principal findings In order to estimate the threat for France due to the introduction of vectors found in animal trucks (cattle or horses) from at-risk countries (Balkans and neighbours), a quantitative import risk analysis (QIRA) model was developed according to the international standard. Using stochastic QIRA modelling and combining experimental/field data and expert opinion, the yearly risk of LSDV being introduced by stable flies (Stomoxys calcitrans), that travel in trucks transporting animals was between 6 x 10−5 and 5.93 x 10−3 with a median value of 89.9 x 10−5 ; it was mainly due to the risk related to insects entering farms in France from vehicles transporting cattle from the at-risk area. The risk related to the transport of cattle going to slaughterhouses or the transport of horses was much lower (between 2 x 10−7 and 3.73 x 10−5 and between 5 x 10−10 and 3.95 x 10−8 for cattle and horses, respectively). The disinsectisation of trucks transporting live animals was important to reduce this risk. Conclusion and significance The development of a stochastic QIRA made it possible to quantify the risk of LSD being introduced in France through the import of vectors that travel in trucks transporting animals. This tool is of prime importance because the LSD situation in the Balkans is continuously changing. Indeed, this model can be updated to process new information on vectors and the changing health situation, in addition to new data from the TRAde Control and Expert System (TRACES, EU database). This model is easy to adapt to different countries and to other vectors and diseases.

La caudofagia representa un problema de gran importancia y está presente en la mayoría de las granjas de explotaciones porcinas intensivas a nivel mundial, afectando de forma negativa el bienestar animal y la productividad. Este estudio se llevó a cabo por medio de revisiones bibliográficas, así como observaciones directas en una granja con una explotación porcina intensiva ubicada en Girona, la cual requirió un análisis situacional acerca del bienestar en sus animales, tomando en cuenta que en el pasado tuvo problemas relacionados con caudofagia; sin embargo, durante el trabajo de campo realizado en dicha granja, no se registraron hallazgos de mordeduras (cola, orejas y laterales). Los principales factores de riesgo de las mordeduras de cola están muy ligados a la falta de material de enriquecimiento y suelos de cemento, mala ventilación (fluctuaciones importantes de temperaturas, concentración de amoníaco elevada), densidades altas, problemas gastrointestinales, salud inadecuada, dieta subóptima o desequilibrada, factores individuales (edad, genética). La caudofagia puede tener efectos adversos sobre las características de la canal y la calidad del producto final, reduciendo el tiempo de crecimiento y cebo asociados a problemas de salud, lo cual supone graves pérdidas económicas para el sector de la carne de cerdo por canales dañadas y decomiso de canales al momento del sacrificio. La producción porcina debe manejarse con mucho conocimiento y especialización, de manera que los encargados o el personal de las instalaciones son fundamentales para actuar de forma oportuna y eficaz, enfocándose en el bienestar animal para la correcta prevención de este mal comportamiento, basados en las normas de densidades máximas permitidas con el propósito de minimizar riesgos de aparición de caudofagia, así como lograr la optimización de los recursos. Caudofagia represents a problem of great importance and is present in most farms of intensive pig farms worldwide, negatively affecting animal welfare and productivity. This study was carried out through bibliographic reviews, as well as direct observations on a farm with an intensive pig farm located in Girona, which required a situational analysis about the welfare of its animals, taking into account that in the past it had problems related to tailbug; however, during the field work carried out on the farm, no bite findings (tail, ears and sides) were recorded. The main risk factors of tail bites are closely linked to the lack of enrichment material and cement floors, poor ventilation (significant temperature fluctuations, high ammonia concentration), high densities and large groups, gastrointestinal problems, inadequate health , suboptimal or unbalanced diet, individual factors (age, genetics). Caudofagia can have adverse effects on the characteristics of the carcass and the quality of the final product, reducing the time of growth and baiting associated with health problems, which supposes serious economic losses for the pork sector by damaged channels and seizure of channels at the time of sacrifice. Swine production must be managed with much knowledge and specialization, so that the managers or the personnel of the facilities are fundamental to act in a timely and effective way, focusing on animal welfare for the correct prevention of this bad behavior, based on the norms of maximum allowed densities with the purpose of minimizing risk of the appearance of tailbusting, as well as achieving the optimization of resources.

Antimicrobials have been used in a prophylactic way to decrease the incidence of digestive disorders during the piglet post-weaning period. Nowadays, it is urgent to reduce their consumption in livestock to address the problem of antimicrobial resistance. In this study, the effect of a product on piglet microbiota has been investigated as an alternative to antimicrobials. Three groups of ten post-weaning pigs were sampled at 0, 15 and 30 days one week post-weaning; the control, antibiotic and feed additive group received a standard post-weaning diet without antibiotics or additives, the same diet as the control group but with amoxicillin and colistin sulphate and the same diet as the control group but with a feed additive (Sanacore-EN, Nutriad International N.V.), respectively. The total DNA extracted from faeces was used to amplify the 16S RNA gene for massive sequencing under manufacturer's conditions. Sequencing data was quality filtered and analyzed using QIIME software and suitable statistical methods. In general terms, age modifies significantly the microbiota of the piglets. Thus, the oldest the animal, the highest bacterial diversity observed for the control and the feed additive groups. However, this diversity was very similar in the antibiotic group throughout the trial. Interestingly, a clear increase in abundance of Bacillus and Lactobacillus spp was detected within the feed additive group versus the antibiotic and control groups. In conclusion, the feed additive group had a positive effect in the endogenous microbiota of post-weaning pigs increasing both, the diversity of bacterial families and the abundance of lactic acid bacteria during the post-weaning period. This work was supported by a research contract of the University of Lleida with Nutriad International N.V. The funder provided support in the form of salaries for authors [Tim Goosens and Alvaro Bermejo], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the "author contributions" section.