Environmental DNA: A promising factor for tuberculosis risk assessment in multi-host settings

[EN] Attaining and maintaining the Official Tuberculosis Free status continues to be a challenge when several domestic and wild hosts contribute to the maintenance of the Mycobacterium tuberculosis complex (MTC). Local tuberculosis hotspots are sometimes identified in cattle in low-prevalence regions. We have, therefore, studied one such hotspot in depth in order to produce an epidemiological diagnosis. Host population size and MTC prevalence were estimated in selected wildlife and in livestock, while on-cattle environmental DNA detection was additionally used as a proxy for risk of exposure at the farm (herd) level. Positive skin test reactors were found on16 of the 24 cattle farms studied in the period 2012-2016. Although all goats tested negative to the skin test during this period, MTC was confirmed in four sheep at slaughter, thus indicating an unknown prevalence of infection in this host species. With regard to wildlife, the prevalence of MTC infection based on culture was 8.8% in the case of wild boar (Sus scrofa), and the only road-killed badger (Meles meles) submitted for culture tested positive. Two criteria were employed to divide the cattle farms into higher or lower risk: tuberculosis testing results and environmental DNA detection. Environmental MTC DNA detection yielded significant differences regarding "use of regional pastures" and "proximity to woodland". This study suggests that on-animal environmental DNA sampling may help when assessing contact risk as regards MTC in livestock at the herd level. This tool opens up new avenues of epidemiological research in complex multi-host settingsSIThis work was supported by the Programa de Tecnologı´as Avanzadas en Vigilancia Sanitaria (TAVS) from the Comunidad de Madrid (ref. S2013/ABI-2747). JMG holds a FPI predoctoral scholarship (BES-2015- 072206), funded by MINECO. This is a contribution to MINECO grant CGL2017-89866 WildDriver and EU-FEDER. This is also a contribution to Valle de Alcudia pilot project SG-2019-02 from PDR-CLM, and to GOSTU project EU-FEADER (AEIAGRI-PNDR-MAPA, ref. 20190020007521). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscrip

Tendance temporelle de la tuberculose dans la communauté d’ongulés du Parc National de Doñana

Trabajo presentado al 35º Encuentro GEEFSM (Groupe d’Etudes sur l’Eco-pathologie de la Faune Sauvage de Montagne), celebrado en Cofrentes, Muela de Cortes (España) del 1 al 4 de junio de 2017.[ES]: El control de las enfermedades infecciosas compartidas entre la fauna silvestre y el ganado es complejo debido al escaso conocimiento de los procesos epidemiológicos que determinan la transmisión y persistencia de patógenos en la comunidad de hospedadores. La tuberculosis animal (TB) causada por la infección crónica del Complejo Mycobacterium tuberculosis (CMT) es endémica en jabalí (Sus scrofa), ciervo (Cervus elaphus) y gamo (Dama dama) en el centro-sur de España. En este contexto es clave conocer la dinámica temporal de la infección y los factores que modulan el mantenimiento de la TB en el medio natural y, consecuentemente, en el ganado en extensivo. Así, en este estudio utilizamos datos de seguimiento sanitario de ungulados silvestres y de bovino del Parque Nacional de Doñana (PND) para evaluar la tendencia temporal en la prevalencia de TB e identificar sus determinantes. Entre 2006 y 2016 se ha realizado la necropsia, el diagnóstico clínico- lesional y confirmación por cultivo microbiológico en un total de 852 jabalíes, 356 ciervos y 338 gamos dentro del plan de vigilancia sanitaria del PND. Por otro lado, la información sanitaria de 1139 cabezas de ganado bovino ha sido facilitada por los servicios veterinarios oficiales. El análisis espacio-temporal de la infección y sus principales factores de riesgo se ha realizado usando sistemas de información geográfica y modelos lineales generalizados. Los resultados muestran prevalencias de TB más elevadas en jabalí (69,44%), seguido de ciervo (43,38%) y gamo (19,88%); mientras que la incidencia en bovino es del 9%. La tendencia temporal de la infección durante el período estudiado es creciente en ungulados silvestres y se mantiene estable en el ganado bovino, a pesar de los intensos esfuerzos de erradicación en esta especie. En general, varios factores individuales, poblacionales, climatológicos y geográficos están involucrados en la dinámica de la enfermedad. Los individuos adultos muestran una prevalencia elevada respecto a los jóvenes en todas las especies, como se ha sugerido previamente en enfermedades de carácter crónico. En cuanto a la distribución espacial de la infección, se mantiene un marcado gradiente Norte- Sur descrito previamente en 2008, observándose valores superiores de prevalencia en aquellas áreas situadas al norte del PND. Esta situación podría estar mediada por las diferencias en la densidad de hospedadores y en el uso de los recursos. Por ejemplo, se aprecia un mayor riesgo relativo de TB en la cercanía a zonas de elevada agregación de individuos, como es el húmedo borde de la marisma. Estos resultados sobre la dinámica de la TB son fundamentales para comprender la epidemiología de la enfermedad en la comunidad de hospedadores y permitirán ayudar en el diseño de futuras estrategias de control.[IT]: Il controllo delle malattie infettive condivise tra la fauna selvatica e il bestiame è complesso a causa della scarsa conoscenza dei processi epidemiologici che determinano la trasmissione e la persistenza dei patogeni nella comunità di animali ospiti. La tubercolosi animale (TB) causata dall’infezione cronica da parte del Complesso Mycobacterium tuberculosis (CMT) è endemica nel cinghiale (Sus scrofa), cervo (Cervus elaphus) e daino (Dama dama) nel centro- sud della Spagna. In questo contesto è essenziale conoscere la dinamica temporale dell’infezione e i fattori che modulano il mantenimento della TB nell’ambiente naturale e, di conseguenza, nell’allevamento estensivo. In questo studio vengono utilizzati dati di controllo sanitario di ungulati silvestri e bovini nel Parco Nazionale della Dognana (PND), per valutare l’evoluzione temporale nella prevalenza della TB e identificarne i fattori determinanti. Tra il 2006 e il 2016 sono stati realizzati necroscopia, diagnosi clinico-lesionale e conferma tramite coltura microbiologica su un totale di 852 cinghiali, 356 cervi e 338 daini nell’ambito del piano di vigilanza sanitaria del PND. Le informazioni relative a 1139 teste di bestiame (bovino) sono state fornite invece dai servizi sanitari veterinari ufficiali. L’analisi spazio-temporale dell’infezione e dei suoi principali fattori di rischio è stata realizzata utilizzando sistemi di informazione geografica e modelli lineari generalizzati. I risultati mostrano prevalenza di TB più alta nelcinghiale (69,44%), seguito da cervo (43,38%) e daino (19,88%), mentre l’incidenza nel bestiame è del 9%. Durante il periodo studiato l’infezione è andata aumentando negli ungulati selvatici e si è mantenuta stabile nel bestiame, nonostante gli intensi sforzi di eradicazione in questa specie. In generale, vari fattori individuali, demografici, climatologici e geografici sono coinvolti nella dinamica della malattia. Gli individui adulti mostrano una prevalenza più elevata rispetto ai giovani in tutte le specie, come si era già visto precedentemente in altre malattie a carattere cronico. In quanto alla distribuzione spaziale dell’infezione, si mantiene una marcata prevalenza a nord rispetto al sud, già descritta prima nel risultando valori superiori nelle aree situate al nord del PND. Questa situazione potrebbe essere dovuta alle differenze di densità demografica degli ospiti e nell’uso condiviso delle risorse. Per esempio, è stato riscontrato un maggiore rischio relativo di TB nella vicinanza di zone con un alto indice di aggregazione degli individui, come ad esempio i terreni paludosi. Questi risultati sulla dinamica della TB sono fondamentali per comprendere l’epidemiologia della malattia nella comunità degli animali che ne sono ospiti e permetteranno lo sviluppo di future strategie di controllo.[FR]: Le contrôle des maladies infectieuses communes à la faune sauvage et à l’élevage est complexe dû aux faibles connaissances des processus épidémiologiques qui déterminent la transmission et la persistance des pathogènes dans la population d’hôtes. La tuberculose animale (TB) causée par l’infection chronique du Complejo Mycobacterium tuberculosis (CMT) est endémique chez le sanglier (Sus scrofa), le cerf (Cervus elaphus) et le daim (Dama dama) dans le centre-sud de l’Espagne. Dans ce contexte, il est primordial de connaître la dynamique temporelle de l’infection et les facteurs qui modulent le maintien de la TB dans le milieu naturel et, par conséquent, dans l’élevage extensif. Ainsi, dans cette étude, nous utilisons des données de suivi sanitaire des ongulés sauvages et des bovins du Parc National de Doñana (PND) pour évaluer la tendance temporelle dans la prévalence de TB et identifier ses déterminants. Entre 2006 et 2016, il a été réalisé l’autopsie, le diagnostic clinique et la confirmation par culture microbiologique sur un total de 852 sangliers, 356 cerfs et 338 daims dans le plan de vigilance sanitaire du PND. D’autre part, l’information sanitaire de 1139 têtes de bovins a été apportée par les services vétérinaires officiels. L’analyse spatio-temporelle de l’infection et ses principaux facteurs de risque a été réalisée en utilisant ses systèmes d’information géographique et des modèles linéaux généraux. Les résultats indiquent des prévalences de TB plus élevées chez le sanglier (69,44%), suivi du cerf (43,38%) et du daim (19,88%) ; alors que l’incidence chez les bovins es de 9%. La tendance temporelle de l’infection durant la période étudiée est croissante chez les ongulés sauvages et reste stable chez les bovins domestiques, malgré les efforts intenses d’éradication dans cette espèce. En général, plusieurs facteurs individuels, de groupes, climatologiques et géographiques sont impliqués dans la dynamique de la maladie. Les individus adultes présentent une prévalence élevée par rapport aux jeunes et ce dans toutes les espèces, ce qui est généralement le cas pour beaucoup de maladies chroniques. Par rapport à la distribution spatiale de l’infection, elle est surtout située dans le Nord-Sud, comme il a été décrit précédemment en 2008, où des valeurs supérieures de prévalence ont été observées dans les aires situées au nord du PND. Cette situation pourrait être due aux différences dans la densité des hôtes et dans l’utilisation des ressources. Par exemple, on observe un risque plus grand de TB à proximité des zones de densité élevée d’individus, comme le bord humide des marais. Ces résultats sur la dynamique de la TB sont fondamentaux pour comprendre l’épidémiologie de la maladie dans la communauté d’hôte et permettent d’aider pour le développement de futures stratégies de contrôle.Peer reviewe

The physiological cost of male-biased parasitism in a nearly monomorphic mammal

Background: Even though male-biased parasitism is common in mammals, little effort has been made to evaluate whether higher parasitic burden in males results in an extra biological cost, and thus a decrease in fitness. Body condition impairment and the augmentation of oxidative stress can be used as indicators of the cost of parasite infections. Here, we examined relationships between gastrointestinal and respiratory helminths, body condition and oxidative stress markers (glutathione peroxidase, paraoxonase-1) in 28 Pyrenean chamois (Rupicapra p. pyrenaica) sampled in autumn. Results: Only male chamois showed a reduction in body condition and higher oxidative stress due to parasite infection, likely because of the extremely high parasite burdens observed in males. Conclusions: This study made evident a disparity in the physiological cost of multiple parasitism between sexes in a wild mammal, mainly due to parasitic richness. Because of the similar life expectancy in male and female chamois, we suggest that males may have developed natural mechanisms to compensate for higher parasite loads during the rut

Mutations in TP53 and JAK2 are independent prognostic biomarkers in B-cell precursor acute lymphoblastic leukaemia

[EN]Background: In B-cell precursor acute lymphoblastic leukaemia (B-ALL), the identification of additional genetic alterations associated with poor prognosis is still of importance. We determined the frequency and prognostic impact of somatic mutations in children and adult cases with B-ALL treated with Spanish PETHEMA and SEHOP protocols. Methods: Mutational status of hotspot regions of TP53, JAK2, PAX5, LEF1, CRLF2 and IL7R genes was determined by next-generation deep sequencing in 340 B-ALL patients (211 children and 129 adults). The associations between mutation status and clinicopathological features at the time of diagnosis, treatment outcome and survival were assessed. Univariate and multivariate survival analyses were performed to identify independent prognostic factors associated with overall survival (OS), event-free survival (EFS) and relapse rate (RR). Results: A mutation rate of 12.4% was identified. The frequency of adult mutations was higher (20.2% vs 7.6%, P=0.001). TP53 was the most frequently mutated gene (4.1%), followed by JAK2 (3.8%), CRLF2 (2.9%), PAX5 (2.4%), LEF1 (0.6%) and IL7R (0.3%). All mutations were observed in B-ALL without ETV6-RUNX1 (P=0.047) or BCR-ABL1 fusions (P<0.0001). In children, TP53mut was associated with lower OS (5-year OS: 50% vs 86%, P=0.002) and EFS rates (5-year EFS: 50% vs 78.3%, P=0.009) and higher RR (5-year RR: 33.3% vs 18.6% P=0.037), and was independently associated with higher RR (hazard ratio (HR)=4.5; P=0.04). In adults, TP53mut was associated with a lower OS (5-year OS: 0% vs 43.3%, P=0.019) and a higher RR (5-year RR: 100% vs 61.4%, P=0.029), whereas JAK2mut was associated with a lower EFS (5-year EFS: 0% vs 30.6%, P=0.035) and a higher RR (5-year RR: 100% vs 60.4%, P=0.002). TP53mut was an independent risk factor for shorter OS (HR=2.3; P=0.035) and, together with JAK2mut, also were independent markers of poor prognosis for RR (TP53mut: HR=5.9; P=0.027 and JAK2mut: HR=5.6; P=0.036). Conclusions: TP53mut and JAK2mut are potential biomarkers associated with poor prognosis in B-ALL patients.European Commision (EC). Funding FP7/SP1/HEALTH. Project Code: 30624

Red deer reveal spatial risks of Crimean-Congo haemorrhagic fever virus infection

In press.Crimean-Congo haemorrhagic fever virus (CCHFV) continues to cause new human cases in Iberia while its spatial distribution and ecological determinants remain unknown. The virus remains active in a silent tick-animal cycle to which animals contribute maintaining the tick populations and the virus itself. Wild ungulates, in particular red deer, are essential hosts for Hyalomma ticks in Iberia, which are the principal competent vector of CCHFV. Red deer could be an excellent model to understand the ecological determinants of CCHFV as well as to predict infection risks for humans because it is large, gregarious, abundant and the principal host for Hyalomma lusitanicum. We designed a cross-sectional study, analysed the presence of CCHFV antibodies in 1444 deer from 82 populations, and statistically modelled exposure risk with host and environmental predictors. The best-fitted statistical model was projected for peninsular Spain to map infection risks. Fifty out of 82 deer populations were seropositive, with individual population prevalence as high as 88%. The highest prevalence of exposure to CCHFV occurred in the southwest of the Iberian Peninsula. Climate and ungulate abundance were the most influential predictors of the risk of exposure to the virus. The highest risk regions were those where H. lusitanicum is most abundant. Eight of the nine primary human cases occurred in or bordering these regions, demonstrating that the model predicts human infection risk accurately. A recent human case of CCHF occurred in northwestern Spain, a region that the model predicted as low risk, pointing out that it needs improvement to capture all determinants of the CCHFV infection risk. In this study, we have been able to identify the main ecological determinants of CCHFV, and we have also managed to create an accurate model to assess the risk of CCHFV infection.This study has been funded by the Spanish Ministry for the Science and Innovation (MCI) through projects CGL2017-89866-R & E-RTA2015-0002-C02-02, and by the Regional Government of Castilla-La Mancha and the European Social Fund (ESF) through project SBPLY/19/180501/000321. RC-M, LC-M, SJ-R and JM-G would like to thank the MCI, ESF and the University of Castilla-La Mancha for their support through contracts PRE2018-083801, PEJ2018-003155-A, 2018/12504 and BES-2015-072206, respectively. BC was supported by FCT-Fundação para a Ciência e Tecnologia (2020.04872.BD). DG-B would like to thank MCI and Instituto de Salud Carlos III (CD19CIII/00011) for funding him.Peer reviewe

Male-biased gastrointestinal parasitism in a nearly monomorphic mountain ungulate

Background: Pyrenean chamois (Rupicapra pyrenaica pyrenaica) is a nearly monomorphic mountain ungulate with an unbiased sex-specific overwinter adult survival. Few differences in gastrointestinal parasitism have been reported by coprology as yet. This study aims to assess diversity, prevalence, intensity of infection and aggregation of gastrointestinal nematodes in male and female adult chamois. We expect no differences in the parasite infection rates between sexes. Findings: Gastrointestinal tracts of 28 harvested Pyrenean chamois in the Catalan Pyrenees (autumn 2012 and 2013) were necropsied and sexual differences in the diversity and structure of parasite community, prevalence, intensity of infection, and richness were investigated. We found 25 helminth species belonging to 13 different genera. Conclusions: Contrary to our expectations, male chamois showed different parasite communities, higher prevalence, intensity of infection and richness than females. Such sexual differences were clear irrespective of age of individuals. Hence, male chamois must cope with a more diverse and abundant parasite community than females, without apparent biological cost. Further research will be required to confirm this hypothesis

Control integrado de la tubercolisis en la interfase entre la fauna silvestre y el ganado doméstico

Resumen del trabajo presentado a las V Jornadas Doctorales de la Universidad de Castilla-La Mancha, celebradas en Ciudad Real (España) el 6 de octubre de 2015.La tuberculosis bovina es una enfermedad crónica que afecta a una amplia variedad de especies domésticas y fauna silvestre, causada por las bacterias del Complejo Mycobacterium tuberculosis, principalmente Mycobacterium bovis y Mycobacterium caprae. Esta enfermedad zoonótica causa graves pérdidas económicas a la industria ganadera, principalmente al ganado bovino, principal reservorio de la enfermedad sobre el que se están realizando grandes esfuerzos de control. Además de los reservorios domésticos, en España se conocen cuatro especies silvestres capaces de actuar como reservorios de infección: el jabalí, el ciervo y con menor relevancia el gamo y el tejón. Así pues, la capacidad de la bacteria para resistir temporalmente en ambientes acuáticos conlleva la potencial transmisión tanto entre el propio ganado como entre este y la fauna silvestre, ya sea por contacto directo o por interacción indirecta al habitar los mismos espacios. En este escenario, el objetivo del presente trabajo es aplicar los conocimientos científicos disponibles para establecer medidas y normas de bioseguridad destinadas a la reducción de las interacciones entre el ganado, el reservorio silvestre y el ambiente. Para ello, se han estado visitando explotaciones ganaderas en las provincias de Ciudad Real, Córdoba, Sevilla, Cádiz, Huelva y Cáceres, con el fin de estudiar detenidamente sus características epidemiológicas y proponer medidas efectivas, prácticas y realistas para el ganadero. Posteriormente se llevará un seguimiento de las propuestas llevadas a término en cada explotación y se monitorizarán para evaluar la efectividad de cada programa personalizado de bioseguridad. Por otra parte, se diseñarán y evaluarán cebos adecuados para la administración efectiva, segura y selectiva de vacunas orales y otros compuestos para ciervos (Cervus elaphus) de todas las edades en condiciones ambientales mediterráneas. De esta forma se aportará conocimiento práctico para el posterior desarrollo de herramientas y protocolos de control de la tuberculosis bovina en rumiantes domésticos y silvestres, basados en vacunas inactivadas. Finalmente, a través de las conclusiones de este trabajo, se desarrollarán y trasladarán al sector ganadero protocolos efectivos de evaluación del riesgo de contacto, como base para implementar programas específicos de mitigación del riesgo de transmisión de enfermedades compartidas en explotaciones ganaderas extensivas del centro y sur de España.Peer reviewe

Riesgos asociados al aporte de alimento concentrado (tacos)

Peer reviewe

Dynamic network of interactions in the wildlife-livestock interface in Mediterranean Spain: An epidemiological point of view

The correct management of diseases that are transmitted between wildlife and livestock requires a reliable estimate of the pathogen transmission rate. The calculation of this parameter is a challenge for epidemiologists, since transmission can occur through multiple pathways. The social network analysis is a widely used tool in epidemiology due to its capacity to identify individuals and communities with relevant roles for pathogen transmission. In the present work, we studied the dynamic network of interactions in a complex epidemiological scenario using information from different methodologies. In 2015, nine red deer, seven fallow deer, six wild boar and nine cattle were simultaneously monitored using GPS-GSM-Proximity collars in Doñana National Park. In addition, 16 proximity loggers were set in aggregation points. Using the social network analysis, we studied the dynamic network of interactions, including direct and indirect interactions, between individuals of different species and the potential transmission of pathogens within this network. The results show a high connection between species through indirect interactions, with a marked seasonality in the conformation of new interactions. Within the network, we differentiated four communities that included individuals of all the species. Regarding the transmission of pathogens, we observed the important role that fallow deer could be playing in the maintenance and transmission of pathogens to livestock. The present work shows the need to consider different types of methodologies in order to understand the complete functioning of the network of interactions at the wildlife/livestock interface. It also provides a methodological approach applicable to the management of shared diseases.The present work has benefitted from the financial aid of a research Grant funded by MINECO (AGL2013-48523-C3-1-R and AGL2016-76358-R). R.T.-O. holds a pre-doctoral scholarship from UCLM. J.A.B. is supported by the EU project (H2020 VACDIVA 862874). J.M.-G. was supported by an FPI grant from MCIU (BES-2015-072206). P.A. received support from UCLM through an extension of Ramón y Cajal contract (RYC-2012-11970).Peer reviewe