Modelling the TB spatial risk in a complex multi-host system assessed by drones

Trabajo presentado a la VI International M. bovis Conference, celebrada en Cardiff (Reino Unido) del 16 al 19 de junio de 2014.The present work has benefited from the financial aid of research grants JCCM (PEII10-0262-7673), MINECO (AGL2010-20730‐C02‐01), Aeromab Project (P07‐RNM‐03246), and the PLANET Project (EU, Cooperation, 7th FP, 2010). J.A.B. holds an FPU pre‐doctoral scholarship. P.A. is supported by MINECO‐UCLM through ‘Ramón y Cajal’ contract (RYC‐2012‐11970) and partly by EMIDA‐ERA‐NET grant APHAEA (219235‐FP7‐ERA‐NET‐EMIDA; www.aphaea.eu). Acknowledgements We would like to thank E. Guerrero and M.A. Aguilar, the pilots and technicians who controlled the UAS and prepared the image mosaic. The present work has benefited from the financial aid of research grants JCCM (PEII10‐0262‐7673), MINECO (AGL2010‐20730‐C02‐01), Aeromab Project (P07‐RNM‐03246), and the PLANET Project (EU, Cooperation, 7th FP, 2010). J.A.B. holds an FPU pre‐doctoral scholarship. P.A. is supported by MINECO‐UCLM through ‘Ramón y Cajal’ contract (RYC‐2012‐11970) and partly by EMIDA‐ERA‐NET grant APHAEA (219235‐FP7‐ERA‐NET‐EMIDA; www.aphaea.eu).Peer Reviewe

Testing Eurasian wild boar piglets for serum antibodies against Mycobacterium bovis

A. Che’ Amat et al.Animal tuberculosis (TB) caused by infection with Mycobacterium bovis and closely related members of the M. tuberculosis complex (MTC), is often reported in the Eurasian wild boar (. Sus scrofa). Tests detecting antibodies against MTC antigens are valuable tools for TB monitoring and control in suids. However, only limited knowledge exists on serology test performance in 2-6 month-old piglets. In this age-class, recent infections might cause lower antibody levels and lower test sensitivity. We examined 126 wild boar piglets from a TB-endemic site using 6 antibody detection tests in order to assess test performance. Bacterial culture (. n=. 53) yielded a M. bovis infection prevalence of 33.9%, while serum antibody prevalence estimated by different tests ranged from 19% to 38%, reaching sensitivities between 15.4% and 46.2% for plate ELISAs and between 61.5% and 69.2% for rapid immunochromatographic tests based on dual path platform (DPP) technology. The Cohen kappa coefficient of agreement between DPP WTB (Wildlife TB) assay and culture results was moderate (0.45) and all other serological tests used had poor to fair agreements. This survey revealed the ability of several tests for detecting serum antibodies against the MTC antigens in 2-6 month-old naturally infected wild boar piglets. The best performance was demonstrated for DPP tests. The results confirmed our initial hypothesis of a lower sensitivity of serology for detecting M. bovis-infected piglets, as compared to older wild boar. Certain tests, notably the rapid animal-side tests, can contribute to TB control strategies by enabling the setup of test and cull schemes or improving pre-movement testing. However, sub-optimal test performance in piglets as compared to that in older wild boar should be taken into account.This is a contribution to Spanish Government MINECO Plan Nacional I+D+I grant AGL2014-56305 and FEDER, to a contract between CDTI and Glenton, and to the EU FP7 grant WildTBvac #613779. Azlan Che Amat has a PhD grant from the Malaysian Government, and José Angel Barasona and Iratxe Diéz-Delgado acknowledge PhD grants from the Spanish Government.Peer Reviewe

Using the FVB strain of mice for the evaluation of clinical and experimental ketamine (IP) associated with phenothiazines, benzodiazepines and α2-agonists

El objetivo de este trabajo ha consistido en evaluar el estado fisiológico de los animales mediante el control de las frecuencias respiratoria y cardiaca así como la tasa de saturación de oxígeno durante la anestesia con ketamina asociada a otros fármacos. Para ello hemos utilizado 40 ratones FVB consanguíneos (20 machos y 20 hembras) de 11 semanas de edad, a los que se les administró por vía intraperitoneal ketamina asociada a un depresor del sistema nervioso central: acepromazina, diazepam, medetomidina, midazolam o xilazina. Obtuvimos resultados que difirieron mucho entre sexos, concluyendo que en machos los mejores resultados obtenidos fueron con la asociación a los α2-agonistas, mientras que en las hembras, al menos a las dosis empleadas, no pudimos afirmar que ninguna de las asociaciones fuese óptima.The aim of this work has been to evaluate the physiological status of animals by controlling the respiratory rate and heart rate and oxygen saturation during anesthesia with ketamine in combination with other drugs. We have used 40 consanguine FVB mice (20 males and 20 females) from 11 weeks of age, who were administered intraperitoneally with a ketamine-associated central nervous system depressant:acepromazina, diazepam, medetomidine, midazolam or xylazine. We obtained results that differed greatly between the sexes, in males, concluding that the best results were obtained with the association of α2- agonists, while in females, at least at the doses employed, did not say that none of the associations were optima

Assessment of an oral Mycobacterium bovis BCG vaccine and an inactivated M. bovis preparation for wild boar in terms of adverse reactions, vaccine strain survival, and uptake by nontarget species

Wildlife vaccination is increasingly being considered as an option for tuberculosis control. We combined data from laboratory trials and an ongoing field trial to assess the risk of an oral Mycobacterium bovis BCG vaccine and a prototype heat-inactivated Mycobacterium bovis preparation for Eurasian wild boar (Sus scrofa). We studied adverse reactions, BCG survival, BCG excretion, and bait uptake by nontarget species. No adverse reactions were observed after administration of BCG (n = 27) or inactivated M. bovis (n = 21). BCG was not found at necropsy (175 to 300 days postvaccination [n = 27]). No BCG excretion was detected in fecal samples (n = 162) or in urine or nasal, oral, or fecal swab samples at 258 days postvaccination (n = 29). In the field, we found no evidence of loss of BCG viability in baits collected after 36 h (temperature range, 11°C to 41°C). Camera trapping showed that wild boar (39%) and birds (56%) were the most frequent visitors to bait stations (selective feeders). Wild boar activity patterns were nocturnal, while diurnal activities were recorded for all bird species. We found large proportions of chewed capsules (29%) (likely ingestion of the vaccine) and lost baits (39%) (presumably consumed), and the proportion of chewed capsules showed a positive correlation with the presence of wild boar. Both results suggest proper bait consumption (68%). These results indicate that BCG vaccination in wild boar is safe and that, while bait consumption by other species is possible, this can be minimized by using selective cages and strict timing of bait deployment.This is a contribution to Plan Nacional I+D+i AGL2011-30041 from the Ministerio de Economía y Competitividad (MINECO), Spain, and FEDER and to EU FP7 grant WildTBvac. B.B.-B. and I.D.-D. were supported by MINECO grants BES-2009-017401 and BES-2012-052490, respectively. J.A.B. was supported by MICINN grant FPU12/00980.Peer Reviewe

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

Evaluation of the clinical evolution and transmission of SARS-CoV-2 infection in cats by simulating natural routes of infection

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the current pandemic disease denominated as Coronavirus Disease 2019 (COVID-19). Several studies suggest that the original source of this virus was a spillover from an animal reservoir and its subsequent adaptation to humans. Of all the different animals affected, cats are one of the most susceptible species. Moreover, several cases of natural infection in domestic and stray cats have been reported in the last few months. Although experimental infection assays have demonstrated that cats are successfully infected and can transmit the virus to other cats by aerosol, the conditions used for these experiments have not been specified in terms of ventilation. We have, therefore, evaluated the susceptibility of cats using routes of infection similar to those expected under natural conditions (exposure to a sneeze, cough, or contaminated environment) by aerosol and oral infection. We have also evaluated the transmission capacity among infected and naïve cats using different air exchange levels. Despite being infected using natural routes and shed virus for a long period, the cats did not transmit the virus to contact cats when air renovation features were employed. The infected animals also developed gross and histological lesions in several organs. These outcomes confirm that cats are at risk of infection when exposed to infected people, but do not transmit the virus to other cats with high rates of air renovation

Long-term determinants of the seroprevalence of the bluetongue virus in deer species in southern Spain

Resumen del póster presentado a la Joint Virtual Conference of the WDA and EWDA, celebrada del 31 de agosto al 2 de septiembre de 2021.Bluetongue is a vector-borne disease affecting domestic and wild ruminants, with a major socioeconomic impact. Endemic circulation of the bluetongue virus serotype 4 (BTV-4) and BTV-1 have occurred in Spain since 2004 and 2007, respectively. However, epidemiological studies have seldom been approached from a long-term perspective in wild ruminants. 881 deer were necropsied as part of the DNP health-monitoring program. Serum samples were tested for antibodies against BTV. We assessed the main drivers of BTV maintenance in the wild ruminant hosts of Doñana National Park using generalised linear mixed models. Red deer displayed statistically significant higher seroprevalences of BTV (SBT; 78.6%) than fallow deer (53.1%). The detection of BTV-1 and BTV-4 by the serum neutralization test in calves suggested the circulation of both serotypes for the study period. For red deer, wet years together with high densities could associate to most suitable conditions for vector borne BTV transmission. Moreover, proximity to permanent pasturelands was associated with higher SBT, which may be mediated by the higher suitability of this habitat to Culicoides. The differences in the ecology and behaviour of deer species influencing the exposure to the vectors could determine the differences found in the SBT patterns. This study evidences the role that deer species may play in the maintenance of BTV in host communities. However, elucidating the epidemiological role deserves further research.Peer reviewe

Long-term determinants of the seroprevalence of the bluetongue virus in deer species in southern Spain

Bluetongue is a vector-borne disease affecting domestic and wild ruminants, with a major socioeconomic impact. Endemic circulation of the bluetongue virus serotype 4 (BTV-4) and BTV-1 have occurred in Spain since 2004 and 2007, respectively. However, epidemiological studies have seldom been approached from a long-term perspective in wild ruminants. A total of 881 deer (red deer and fallow deer) were necropsied from 2005 to 2018 as part of the DNP health-monitoring program. Serum samples were tested for antibodies against BTV with the aims of assessing the temporal trend and to evaluate the modulating factors: individual, populational, environmental, and stochastic. Red deer displayed statistically significant higher seroprevalences of BTV (SBT; 78.6 ± 3.8%) than fallow deer (53.1 ± 4.7%). The detection of BTV-1 and BTV-4 by the serum neutralization test in calves suggested the circulation of both serotypes over the study period. For red deer, wet years together with high densities could provide suitable conditions for vector borne BTV transmission. Moreover, proximity to high suitability habitat for Culicoides, permanent pasturelands, was associated with higher SBT. The differences in the ecology and behaviour of deer species influencing the exposure to the vectors could determine the differences found in the SBT patterns. This study evidences the role that deer species may play in the maintenance of BTV, however, elucidating the epidemiological role of host in different contexts as well as the consequences of climate change on the competent vector populations and its potential effect on the dynamics of BTV infection in hosts communities deserve further research.This research was funded by the Ministerio de Economía y Competitividad (MINECO; AEI/FEDER, UE; AGL2016-76358-R). S.J. is co-supported by the UCLM and the European Social Fund (2018/12504). P.P. received support from the MINECO (FPU/16/00039).Peer reviewe

Trained immunity is not universal: oral heat-inactivated Mycobacterium bovis confers no protection against the non-enveloped Porcine Circovirus 2

[Background] Trained immunity, the enhanced response of innate cells leading to an improved innate immune response, and antibodies against the glycan galactose-α-1,3-galactose (α-Gal), produced by animals unable to synthesize α-Gal epitopes, have been suggested to provide the host certain advantage in infections with enveloped viruses. Conversely, the evidence of protection against non-enveloped viruses attributed to the referred mechanisms remains scarce. Aiming to evaluate whether a heat-inactivated Mycobacterium bovis (HIMB) immunostimulant, which had proven to protect against related and non-related pathogens, confers an advantage against non-enveloped viruses, we performed an immunization and challenge experiment with porcine circovirus 2 (PCV-2) in swine. Sixteen piglets were randomly assigned to the immunized group (n = 8), which received two oral doses of HIMB with an interval of three weeks, or to the control group (n = 8). All animals were infected by intranasal inoculation with PCV-2 21 days later and euthanized at day 21 post-challenge.[Results] No differences in body weight and body temperature, viremia and viral burden in target tissues, antibody production and histopathological changes in target tissues were observed between the immunized and the control group. Overall, oral immunization with HIMB did not protect pigs against PCV-2 infection.[Conclusions] Our study suggests that HIMB confers no advantage against pathogens lacking α-Gal, mainly non-enveloped viruses such as PCV-2, in α-Gal-producing hosts, such as the swine.The present study has been funded by project MYCOTRAINING SBPLY/19/180501/000174 (Junta de Castilla-La Mancha, Spain, and EU-FEDER). E. Ferreras-Colino (2020/3836) and R. Vaz-Rodrigues (2022/20675) were supported by the predoctoral contract from Universidad de Castilla-La Mancha (UCLM), co-financed by the European Social Fund (ESF). Marinela Contreras was supported by the Ministerio de Ciencia, Innovación y Universidades, Spain, grant IJC2020-042710-I.Peer reviewe