Situación epidemiológica de Pestivirus en el Parque Nacional de Doñana: evaluación de la interacción entre rumiantes domésticos y silvestres

Premio extraordinario de Trabajo Fin de Máster curso 2015-2016. Medicina, Sanidad y Mejora AnimalEn la Península Ibérica se ha descrito una amplia variedad de escenarios epidemiológicos en relación a los Pestivirus (familia Flaviviridae) de rumiantes, Virus de la Diarrea Vírica Bovina -VDVB- y Virus de la Enfermedad de la Frontera -VEF- en poblaciones de rumiantes domésticos y silvestres. Sin embargo, la información relativa al papel de los rumiantes silvestres como reservorios naturales de los Pestivirus en los ecosistemas mediterráneos sigue siendo muy limitada. Durante los meses de octubre y noviembre de 2015 se obtuvieron muestras de suero de 138 bovinos mantenidos en extensivo en el Parque Nacional de Doñana (PND), así como muestras de 101 ciervos (Cervus elaphus) y 102 gamos (Dama dama) que comparten hábitat con el ganado bovino analizado. La presencia de anticuerpos específicos frente a la proteína p80 de Pestivirus se evaluó mediante ELISA de bloqueo (bELISA) y tests de seroneutralización vírica (TSV) (cepas VDVB-1, VDVB-2 y VEF-4). Todas las muestras de suero se analizaron para detectar la presencia de ARN de Pestivirus mediante RT-PCR. El 2,2% (3 de 138) de los bovinos analizados fueron positivos a ELISA y TSV (mayores títulos frente a VDVB-1) mientras que todos los rumiantes silvestres resultaron seronegativos. No se detectó ARN vírico en ninguna de las muestras de suero analizadas. La prevalencia obtenida indica una limitada circulación de Pestivirus en el ganado bovino en la zona de estudio así como la ausencia de circulación en los rumiantes silvestres analizados. Los resultados sugieren que los rumiantes silvestres no representan actualmente un riesgo en la transmisión de Pestivirus para el ganado en el PND. Sin embargo, la presencia de poblaciones no protegidas podría favorecer la aparición de brotes y animales persistentemente infectados tras la introducción de un Pestivirus en el PND.In the Iberian Peninsula a wide range of epidemiological contexts has been described in relation to the ruminant Pestivirus (Flaviviridae family), such as the Bovine Viral Diarrhoea Virus -BVDV- and the Border Disease Virus -BDV- in domestic and wild ruminant populations. However, the information of the wild ruminants‟ role as natural reservoirs of Pestivirus in Mediterranean ecosystems is still limited. During the months of October and November 2015, serum samples were obtained from 138 extensive cattle from Donana National Park (DNP), as well as samples from 101 deer (Cervus elaphus) and 102 fallow deer (Dama dama) sharing habitat with the analyzed cattle. The presence of specific antibodies against p80 protein of Pestivirus was evaluated by using both blocking ELISA (bELISA) and serum neutralization tests (SNT) (BVDV-1, BVDV-2 and BDV-4 strains). All serum samples were analyzed in order to detect the presence of Pestivirus RNA by RT-PCR. A 2.2% (3 out of 138) of the tested cattle was positive to both bELISA and SNT (higher titers against BVDV-1) while all wild ruminants were seronegative. No viral RNA was detected in any of the analyzed serum samples. The obtained prevalence indicates a limited widespread of Pestivirus in cattle inside the studied area, as well as the absence of viral circulation in the wild ruminants performed. The results suggest that currently, wild ruminants do not represent a risk of Pestivirus transmission for livestock in DNP. However, the presence of naïve populations may favor the appearance of outbreaks and persistently infected animals after the introduction of a Pestivirus in the DNP

Identifying environmental drivers of benthic diatom diversity: the case of Mediterranean mountain ponds

[EN] This study aims at elucidating the environmental factors controlling benthic diatom diversity and uniqueness in Mediterranean mountain ponds. Samples of periphytic diatoms were collected in 45 ponds in Andalusia, south of Spain, and analysed by standard methods. Data analysis reveals that diatom diversity is mainly controlled by elevation and hydroperiod. Contrary to the usual findings in the literature, the highest scores on Shannon’s diversity index were found in high-elevation temporary ponds, but this effect is hidden by lake clustering in the analysed dataset. Significant distance-decay similarity (DDS) trends were detected in the analysis of floristic composition among the samples, stressing the importance of spatial factors that may override the effect of other abiotic factors. These findings highlight the role of isolation and dispersal limitation in the configuration of the biogeographical patterns of benthic diatomsSIThis work is part of the project ``Andalusian mountain wetlands: inventory, typologies and conservation'' which is funded by the Center for Advanced Studies in Earth Sciences (CEACTierra) at the University of Jaén. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscrip

Epidemiological survey and risk factors associated with hepatitis E virus in small ruminants in southern Spain

Autochthonous cases of hepatitis E (HE) associated with zoonotic genotypes HEV-3 and HEV-4 have significantly increased in industrialized countries over the last decade. Suidae are generally recognized as the main reservoirs of these genotypes. Susceptibility to HE virus (HEV) infection and zoonotic potential have also been confirmed in other species, including sheep and goat. However, the information about their role in the epidemiology of HEV remains very scarce. The objective of this study was to assess the prevalence, spatial distribution and risk factors associated with HEV exposure in sheep and goats in southern Spain, the country with the highest census of small domestic ruminants in the European Union. Blood samples from 240 sheep and 240 goats were collected between 2015 and 2017. Sera were analysed in parallel using a commercial double-antigen ELISA and real-time PCR. A total of 38 (7.9%; 95%CI: 5.5–10.3) out of 480 sampled animals showed anti-HEV antibodies. By species, the seroprevalences found in sheep and goats were 2.1% (5/240; 95%CI: 0.3–3.9) and 13.8% (33/240; 95%CI: 9.4–18.1) respectively. Anti-HEV antibodies were found on 19 (59.4%; 95%CI: 42.4–76.4) of the 32 sampled farms. The GEE model showed that species (goat) and number of small ruminants in the farm (≤348 animals and ≥538 animals) were risk factors potentially associated with HEV exposure in small ruminants in the study area. HEV RNA was not detected in any of the 480 (0.0%; 95%CI: 0.0–0.8) tested animals. Our results confirm that sheep and goats are naturally, but not equally exposed to HEV and indicate the widespread spatial distribution of HEV among small ruminant populations in southern Spain. Further studies are required to elucidate the role of sheep and goat in the epidemiology of HEV and their potential implications for public health

Taxonomy and morphology of Craticula gadorensis sp. nov. (Bacillariophyta, Stauroneidaceae)

[ES] Introducción y objetivos: Se describe la diatomea Craticula gadorensis sp. nov. a partir de muestras de algas epilíticas recogidas en una balsa ganadera de la Sierra de Gádor, en la provincia de Almería. M&M: Las muestras se tomaron y procesaron siguiendo los protocolos metodológicos para diatomeas. Los caracteres morfológicos y ultraestructurales de este nuevo taxón se describen e ilustran mediante imágenes de microscopía óptica y electrónica. Resultados: La nueva especie se caracteriza por presentar valvas elípticolanceoladas a rómbico-lanceoladas, con ápices ligeramente prolongados, subrostrados, con un área axial lineal, estrecha y un área central un poco dilatada, lineal-lanceolada. Conclusiones: La comparación morfológica con otras especies del género muestra que la combinación de caracteres de Craticula gadorensis (principalmente las dimensiones, la densidad de areolas y la forma del área central) es única[EN] Background and Aims: The diatom Craticula gadorensis sp. nov. is described from samples of epilithic algae collected in a shallow pond from the Gador mountains, in the province of Almería (Spain). M&M: Samples were taken and processed following the usual protocols for diatoms. Morphological and ultrastructural characteristics of this new taxon are described and illustrated using light and scanning electron microscopy. Results: The new species is characterised by having elliptic-lanceolate to rhombiclanceolate valves, with slightly protracted, subrostrate apices, a linear, narrow axial area and a slightly dilated, linear-lanceolate central area. Conclusions: A morphological comparison with related species shows that the combination of features in Craticula gadorensis is unique (mainly the dimensions, areola density and the shape of the central area)SIEste trabajo se enmarca en el proyecto “Humedales de montaña de Andalucía: inventario, tipologías y conservación” financiado por el Centro de Estudios Avanzados en Ciencias de la Tierra (CEACTierra) de la Universidad de Jaé

Monitoring of Schmallenberg virus in Spanish wild artiodactyls, 2006-2015

Schmallenberg disease is an emerging disease that affects domestic and wild ruminants in Europe. An epidemiological survey was carried out to assess exposure to Schmallenberg virus (SBV) in wild artiodactyls in Spain between 2006 and 2015. A total of 1751 sera from wild artiodactyls, including 1066 red deer, 304 fallow deer, 192 mouflon, 109 wild boar, 49 roe deer and 31 Spanish ibex were tested for antibodies against SBV by ELISA and confirmed by virus neutralization test. SBV was not detected between the 2006/2007 and the 2010/2011 hunting seasons. Overall seroprevalence (including samples collected between the 2011/2012 and 2014/2015 hunting seasons) was 14.6% (160/1099; 95%CI: 12.7-16.6). Mean SBV seroprevalence was 13.3±2.6% in red deer, 23.9±4.2% in fallow deer, 16.4±6.1% in mouflon and 2.8±3.1% in wild boar. No antibodies against SBV were found in roe deer or Spanish ibex. The presence of SBV RNA was confirmed in three of 255 (1.2%) spleen samples from wild ruminants analysed by rRT-PCR. In a multivariate mixed-effects logistic regression model, the main risk factors associated with SBV seroprevalence were: species (fallow deer, red deer and mouflon), age (adults) and interactions between hunting areas of more than 1000 hectares and hunting season (2012/2013, 2013/2014 and 2014/2015). The hypothesis of endemic circulation of SBV in the last few years is supported by the detection of SBV RNA in animals sampled in 2011 and 2015, as well as antibodies detected at low level in juveniles in 2012, 2013 and 2014. The results indicate that SBV circulated in wild ruminant populations in Spain during the same period when the virus was first reported in northern Europe, and at least five months before the first case was officially reported in livestock in Spain

Evaluation of two enzyme-linked immunosorbent assays for diagnosis of bluetongue virus in wild ruminants

Bluetongue (BT) is a reportable re-emerging vector-borne disease of animal health concern. Enzyme-linked immunosorbent assays (ELISA) are frequently used in BT surveillance programs in domestic ruminants, but their diagnostic accuracy has not been evaluated for wild ruminants, which can play an important role as natural reservoirs of bluetongue virus (BTV). The aim of this study was to assess two commercial ELISAs for BT diagnosis in wild ruminants using control sera of known BTV infection status and field samples. When control sera were tested, the double recognition ELISA (DR-ELISA) showed 100 % sensitivity (Se) and specificity (Sp), while the competitive ELISA (C-ELISA) had 86.4 % Se and 97.1 % Sp. Using field samples, the selected latent-class analysis model showed 95.7 % Se and 85.9 % Sp for DR-ELISA, 58.2 % Se and 95.8 % Sp for C-ELISA and 84.2 % Se for the serum neutralization test (SNT). Our results indicate that the DR-ELISA may be a useful diagnostic method to assess BTV circulation in endemic areas, while the C-ELISA should be selected when free-areas are surveyed. The discrepancy between control and field samples point out that the inclusion of field samples is required to assess the accuracy of commercial ELISAs for the serological diagnosis of BTV in wild ruminants.info:eu-repo/semantics/acceptedVersio

Description of the first Schmallenberg disease outbreak in Spain and subsequent virus spreading in domestic ruminants

Schmallenberg disease (SBD) is an emerging disease transmitted mainly among ruminant species by biting midges of the genus Culicoides. Since the Schmallenberg virus (SBV) was first identified in Germany in late 2011, it rapidly spread to other European countries. The aims of the present study were to describe the first SBD outbreak in Spain and to assess the spread and risk factors associated with SBV infection in domestic ruminants from nearby farms during the following year. In March 2012, one malformed stillborn lamb from a sheep farm located in Cordoba province (Southern Spain) was subjected to necropsy. Pathological compatible lesions and molecular analyses confirmed the first SBV infection in Spain. Afterwards, serum samples from 505 extensively reared domestic ruminants from 29 farms were analysed using both blocking ELISA and virus neutralization test against SBV. The overall seroprevalence was 54.4% (CI95%: 50.0–58.7). Antibodies were detected in 70.6%, 46.0% and 34.8% of cattle, sheep and goats, respectively. A generalized estimating equation model indicated that the main risk factors associated with SBV infection were: species (cattle), age (adult), and absence of animal insecticide treatment. Pathological and molecular results confirmed the presence of SBV in Spain few months after it was firstly identified in Germany. The seroprevalence detected indicates a widespread circulation of SBV in nearby domestic ruminant farms one year after this first outbreak was reported in Spain. Further studies are warranted to determine the spatio-temporal trend of SBV in domestic ruminants in this country.info:eu-repo/semantics/acceptedVersio

Metal-induced abnormalities in diatom girdle bands

P. 1-14There have been a number of studies that described a serial of type of teratology occurring in different diatom taxa and that highlight the relation between metal concentration and diatom deformities, but this subject still remain not deeply understood. The present study refers to the effect of metal pollution on the diatom Achnanthidium minutissimum s.l. by describing a new form of teratology. The samples were collected in a mine area, Rosia Montana, from Romania. We observed that, exposed to environmental stress, the frustule of diatom cells appeared altered in several ways, with abnormal forms occurring in different diatom species that presented deformed valve outlines, modifications of the raphe canal system, irregular striation or mixed teratologies. In a particular sampling location where A. minutissimum s.l. was identified as the dominant species, 20.53% of the individuals presented an unreported type of deformity. This kind of teratology affects the cingulum, the valvocopula more exactly, by becoming markedly undulat

Distribution of Pestivirus exposure in wild ruminants in Spain

A large‐scale study was carried out to determine the prevalence of antibodies against Pestivirus species in wild ruminants and describe their spatial variation in mainland Spain. Serum samples of 1,874 wild ruminants from different regions of this country were collected between the years 2000 and 2017. A total of 6.6% (123/1,874) animals showed antibodies against Pestivirus by both blocking ELISA (bELISA) and virus neutralization tests (VNT). The prevalence of antibodies against pestiviruses was different both among species and regions. Seroprevalence by species was 30.0% (75/250) in Southern chamois (Rupicapra pyrenaica), 7.0% (25/357) in fallow deer (Dama dama), 2.5% (10/401) in red deer (Cervus elaphus), 2.4% (8/330) in Iberian wild goat (Capra pyrenaica), 1.1% (4/369) in roe deer (Capreolus capreolus) and 0.8% (1/130) in mouflon (Ovis aries musimon), not detecting seropositivity (0/37) in Barbary sheep (Ammotragus lervia). The results confirm that exposure to pestiviruses was detected throughout mainland Spain, with significantly higher seroprevalence in Northern regions associated with the presence of Southern chamois. This indicates an endemic circulation of pestiviruses in Southern chamois and a limited circulation of these viruses in the remaining wild ruminant species during the last two decades, thus suggesting that non‐chamois species are not true Pestivirus reservoirs in Spain. Nonetheless, the high spatial spread of these viruses points out that new epidemic outbreaks in naïve wild ruminant populations or transmission to livestock may occur, evidencing the usefulness of monitoring pestiviruses in wild ruminants, especially at the wildlife–livestock interface.info:eu-repo/semantics/acceptedVersio

Tomato plant extract (Lycopersicon esculentum) obtained from agroindustrial byproducts and its antifungal activity against Fusarium spp.

Phytopathogenic fungi are a constant danger in the production of different crops around the world, especially in melons, since they can cause significant economic losses during the harvest, affecting the quality and shelf life. In recent years, producers have increasingly used chemical pesticides indiscriminately, causing environmental problems and damage to public health. For this reason, phytopathogenic fungi become more resistant. However, it is essential to guarantee the safety, quality, and shelf life of food after harvest, during transportation, storage and marketing. The presence of fungi in food can cause diseases transmitted through the production of toxins. Most producers depend on the discriminated use of chemical pesticides, which is a great challenge to guarantee food safety and sustainable agricultural production. To solve this problem, some extracts derived from tomato plants after harvest containing bioactive compounds have been implemented. These compounds can be natural antifungal agents as they contain phenols, flavonoids, and vitamins. Bioactive compounds emerge as a sustainable and safe opportunity in the search for new antifungal and antimicrobial agents. Therefore, the objective of this study was to determine the in vitro antifungal activity of whole tomato plant extracts on three phytopathogenic fungi. The research findings indicated that a concentration of 74.7 μg/mL of TPE resulted in a complete inhibition of mycelial growth in Fusarium oxysporum, Fusarium graminearum, and Fusarium verticillioides. Additionally, TPE exhibited both fungistatic and fungicidal effects on these Fusarium species, with a MIC50 of 30.7, 31.5, and 29.5, and a MFC of 82.4, 78.6, and 75.8 μg/mL, respectively. As a result, this study suggests that TPE can be considered as an environmentally friendly solution for extracting tomato plants, which can be applied to the surface of whole fruits or incorporated into semi-processed foods