In vitro amplification of BVDV field strains isolated in Argentina: effect of cell line and culture conditions

The aim of this work was to study the in vitro amplification of BVDV (Pestivirus, Flaviridae) field isolates from Argentina in MDBK, BoTur and BHK-21 continuous cell lines. Field isolates 99/134 (mucosal disease), 00/693 (mucosal disease), 04P7016 (respiratory disease) and 04/89 (mucosal disease), genotype 1b, were used and compared with the Singer and NADL reference strains, genotype 1a. Additionally, cell lines derived from explants of bovine testis (RD-420), bovine uterus (NCL-1) and porcine kidney (PKZ) were tested as alternative substrates for BVDV propagation in vitro. The effect of cell line, harvest time and infection protocol was evaluated. The viral titers observed depended on the virus and harvest time but not on the infection protocol. We found that MDBK and BoTur cell lines were susceptible to the infection whereas BHK-21 and PKZ were not. NADL viral titers, 00/693 and 04/89, increased from 24 to 48 h p.i. in BoTur cells and then reached a plateau, whereas those of 99/134 and 04P7016 remained constant between 24 and 72 h p.i. BVDV Singer, on the other hand, presented a maximum titer at 24 h p.i. and then decreased. BVDV-NADL titers increased in MDBK and NCL-1 but not in RD-420 between 24 and 48 h p.i., and then decreased at 72 h p.i. These facts lead us to conclude that neither the subgenotypes (1a, 1b) nor the clinical symptoms of the animal from the virus had been isolated seem to affect the virus cell line kinetics of viral replication in vitro. On the other hand, the most homogenous behavior, the most similar replication curves, and highest titers observed in MDBK and NCL-1 seem to indicate that these lines are generally more susceptible to BVDV replication.The aim of this work was to study the in vitro amplification of BVDV (Pestivirus, Flaviridae) field isolates from Argentina in MDBK, BoTur and BHK-21 continuous cell lines. Field isolates 99/134 (mucosal disease), 00/693 (mucosal disease), 04P7016 (respiratory disease) and 04/89 (mucosal disease), genotype 1b, were used and compared with the Singer and NADL reference strains, genotype 1a. Additionally, cell lines derived from explants of bovine testis (RD420), bovine uterus (NCL-1) and porcine kidney (PKZ) were tested as alternative substrates for BVDV propagation in vitro. The effect of cell line, harvest time and infection protocol was evaluated. The viral titers observed depended on the virus and harvest time but not on the infection protocol. We found that MDBK and BoTur cell lines were susceptible to the infection whereas BHK-21 and PKZ were not. NADL viral titers, 00/693 and 04/89, increased from 24 to 48 h p.i. in BoTur cells and then reached a plateau, whereas those of 99/134 and 04P7016 remained constant between 24 and 72 h p.i. BVDV Singer, on the other hand, presented a maximum titer at 24 h p.i. and then decreased. BVDV-NADL titers increased in MDBK and NCL-1 but not in RD-420 between 24 and 48 h p.i., and then decreased at 72 h p.i. These facts lead us to conclude that neither the subgenotypes (1a, 1b) nor the clinical symptoms of the animal from the virus had been isolated seem to affect the virus cell line kinetics of viral replication in vitro. On the other hand, the most homogenous behavior, the most similar replication curves, and highest titers observed in MDBK and NCL-1 seem to indicate that these lines are generally more susceptible to BVDV replication.Fil: Odeón, Anselmo Carlos. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Leunda, Maria Rosa. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Faverin, Claudia. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Boynak, Natalia. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Vena, M. M.. Biogénesis-Bagó S. A.; ArgentinaFil: Zabal, O.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentin

Diet of 0+ brown trout (Salmo trutta L., 1758) from the river Erro (Navarra, north of Spain)

The diet composition of 41 0+ brown trout (Salmo trutta L., 1758) (33-97 mm TL) captured in August 2002 in the Erro River (North of Spain) is described. The diet was mainly composed by aquatic invertebrates. Excluding nematodes because they are possible parasites, the most consumed prey items were mayflies, dipterans, and crustaceans. Fingerling brown trout refused Chironomidae, Elmidae, and terrestrial invertebrates from the drift, and Elmidae, Heptageniidae, and Leuctridae from the benthos, while they showed preference for Rhyacophilidae from the benthos. Fingerling brown trout seems to act as an opportunistic predator, and the consumption of different preys seems to be influenced by their accessibility, predation risk, and their energetic value.Se analizó la dieta de 41 alevines de trucha común (Salmo trutta L., 1758) (33-97 mm LT) del río Erro (Norte de España). Su alimentación estuvo constituida principalmente por invertebrados acuáticos. Si no se tiene en cuenta a los nematodos por la posibilidad de tratarse de parásitos, las presas más consumidas fueron efémeras, dípteros y crustáceos. Los alevines de trucha común rechazaron los quironómidos, élmidos y los invertebrados terrestres de la deriva, así como los élmidos, heptagénidos y leúctridos del bentos, mientras que mostraron preferencia por los riacofílidos del bentos. Los alevines de trucha común parecen comportarse como depredadores oportunistas, pareciendo estar el consumo de las diferentes presas influenciado por su accesibilidad, el riesgo de predación y el valor energético de cada presa

Upstream migration of anadromous and potamodromous brown trout: patterns and triggers in a 25-year overview

Producción CientíficaRiver fragmentation and alterations in flow and thermal regimes are the main stressors affecting migrating fish, which could be aggravated by climate change and increasing water demand. To assess these impacts and define mitigation measures, it is vital to understand fish movement patterns and the environmental variables affecting them. This study presents a long-term (1995–2019) analysis of upstream migration patterns of anadromous and potamodromous brown trout in the lower River Bidasoa (Spain). For this, captures in a monitoring station were analyzed using Survival Analysis and Random Forest techniques. Results showed that most upstream movements of potamodromous trout occurred in October–December, whereas in June–July for anadromous trout, although with differences regarding sex and size. Both, fish numbers and dates varied over time and were related to the environmental conditions, with different influence on each ecotype. The information provided from comparative studies can be used as a basis to develop adaptive management strategies to ensure freshwater species conservation. Moreover, studies in the southern distribution range can be crucial under climate warming scenarios, where species are expected to shift coldwards.Proyecto de innovación docente de la Universidad de Valladolid (PIF-UVa 2017)Ministerio de Ciencia, Innovación y Universidades (grant PTQ2018-010162)Publicación en abierto financiada por el Consorcio de Bibliotecas Universitarias de Castilla y León (BUCLE), con cargo al Programa Operativo 2014ES16RFOP009 FEDER 2014-2020 DE CASTILLA Y LEÓN, Actuación:20007-CL - Apoyo Consorcio BUCL

International Standardization of Common Names for Iberian Endemic Freshwater Fishes

Las especies de peces dulceacuícolas endémicas de la península Ibérica carecen de nombres comunes en inglés, lo cual frecuentemente causa inconvenientes a los autores en el momento de publicar para una audiencia internacional. Con el objetivo de llenar este vacío, se presenta una lista actualizada de las especies de peces dulceacuícolas endémicas de la península Ibérica con una propuesta razonada de designación internacional estandarizada junto con los nombres comunes en español y/o portugués adoptados en los Libros Rojos NacionalesIberian endemic freshwater fishes do not have standardized common names in English, which is usually a cause of inconveniences for authors when publishing for an international audience. With the aim to tackle this problem, an updated list of Iberian endemic freshwater fish species is presented with a reasoned proposition of a standard international designation along with Spanish and/or Portuguese common names adopted in the National Red Data BooksOs peixes dulciaquícolas endémicos da Península Ibérica näo possuem um nome comum devidamente padronizado em Inglês, o que causa problemas aos investigadores quando publicam em revistas com uma audiência internacional. O presente trabalho procurou resolver esta questäo, incluindo uma lista actualizada das espécies piscícolas endémicas da Península Ibérica e uma proposta fundamentada de nomes comuns em Inglês, juntamente com as designaçöes comuns em Espanhol e/ou Português adoptadas nos respectivos Livros Vermelhos Nacionai

Advantages and insights from a hierarchical Bayesian growth and dynamics model based on salmonid electrofishing removal data

Growth is a fundamental ecological process of stream-dwelling salmonids which is strongly interrelated to critical life history events (emergence, mortality, sexual maturity, smolting, spawning). The ability to accurately model growth becomes critical when making population predictions over large temporal (multi-decadal) and spatial (meso) scales, e.g., investigating the effect of global change. Body length collection by removal sampling is a widely-used practice for monitoring fish populations over such large scales. Such data can be efficiently integrated into a Hierarchical Bayesian Model (HBM) and lead to interesting findings on fish dynamics. We illustrate this approach by presenting an integrated HBM of brown trout (Salmo trutta) growth, population dynamics, and removal sampling data collection processes using large temporal and spatial scales data (20 years; 48 sites placed along a 100 km latitudinal gradient). Growth and population dynamics are modelled by ordinary differential equations with parameters bound together in a hierarchical structure. The observation process is modelled with a combination of a Poisson error, a binomial error, and a mixture of Gaussian distributions. Absolute fit is measured using posterior predictive checks, which results indicate that our model fits the data well. Results indicate that growth rate is positively correlated to catchment area. This result corroborates those of other studies (laboratory, exploratory) that identified factors besides water temperature that are related to daily ration and have a significant effect on stream-dwelling salmonid growth at a large scale. Our study also illustrates the value of integrated HBM and electrofishing removal sampling data to study in situ fish populations over large scales

Millennial land use explains modern high‐elevation vegetation in the submediterranean mountains of Southern Europe

Aim Pinewood decline and scrubland expansion are major features of Late Holocene vegetation history in the Cantabrian Range. However, the drivers of this remarkable vegetation shift remain to be investigated. Here, we aim at disentangling the role of past land use and climate in shaping the high-elevation Cantabrian landscape during the past two millennia. Location Cantabrian Range (northern Iberia). Taxa Pinus sylvestris, Betula, Ericaceae, Juniperus, Poaceae. Methods We conducted high-resolution multiproxy palaeoecological analyses (pollen, plant macrofossils, microscopic charcoal and dung fungi) on lake sediments from Lago del Ausente to reconstruct vegetation, fire occurrence and grazing through time. The chronology is based on 14C (terrestrial plant macrofossils) and 210Pb dating, and Bayesian age-depth modelling (‘rbacon’). We carried out cross-correlation analysis to quantify vegetation responses to fire. Results Between 250 and 900 CE, the vegetation above 1700 m a.s.l. consisted of subalpine scrubland and scattered P. sylvestris trees/stands. Pinewoods with Betula were widespread at slightly lower elevation. This vegetation was resilient to moderate fire disturbance associated with limited pastoral activities. In contrast, enhanced fire occurrence alongside heavier pastoralism led to the demise of pinewoods and their replacement with Betula stands, subalpine scrublands, and meadows between 900 and 1100 CE. Later, the subalpine scrubland-birch tree line did not respond to Little Ice Age cooling. However, further intensification of transhumant herding between 1300 and 1860 CE (‘La Mesta’) triggered birch decline and the establishment of the modern treeless landscape. Main conclusions The extant high-elevation vegetation of the Cantabrian Range is largely the legacy of intensive land use starting more than one millennium ago. Recurrent and severe fires to promote pasturelands led to the regional extirpation of the previously widespread Pinus sylvestris. Future management should aim at preserving the valuable cultural open landscape of mountain scrubland and meadows and also at restoring patches of ancient pine-birch woodlands

Genomic analysis of bovine herpesvirus type 4 (BoHV-4) from Argentina: High genetic variability and novel phylogenetic groups

Bovine herpesvirus 4 (BoHV-4) is a γ-herpesvirus that has been isolated both from apparently healthy animals and from cattle with a variety of clinical signs, including post-partum endometritis and abortion. BoHV-4 causes either a persistent or a latent infection in cells of the monocyte/macrophage lineage. Two groups of BoVH-4 strains have been defined based on their restriction patterns: the Movar-like strains (European prototype) and the DN 599-like strains (American prototype). The purpose of the present study was to genetically characterize wild type BoHV-4 strains isolated from vaginal discharges of aborted cows in Argentina. The virus was identified by isolation and nested PCR in all vaginal discharge samples from aborted cows, either as a sole agent or in association with other pathogens. Restriction enzyme profiling and phylogenetic analysis demonstrated that there is a high genetic variability among the studied field isolates. The existence of three groups of strains, which were designated as genotypes 1, 2 and 3, is described. Genotypes 1 and 2 possibly correspond to the Movar-like and DN 599-like groups, respectively, whereas Genotype 3 corresponds to a novel group. Two viral strains did not cluster into any of these three groups, indicating that other genotypes could be circulating in Argentina. These results suggest a complex epidemiological background for the Argentinean BoHV-4 strains, probably influenced by independent events of genetic drift. This hypothesis cannot be rejected based on the available data. However, there is no direct evidence supporting this possibility. Thus, it seems speculative to suggest that interspecific jumps are responsible for the observed phylogenetic grouping. On the other hand, our analyses suggest a geographical structure for the observed viral genotypes, since genotypes 1 and 2 included the European (Movar-like) and American (DN599-like) reference strains, respectively. Geographic dispersion is known to be a driver of herpes viruses diversification, and independent evolution in geographical isolated places ensures the emergence of particular mutations in each location due to genetic drift (Compans, 2007; Zong et al., 1999). Therefore, at this point, the genetic drift hypothesis is the one that requires less ad-hoc considerations and thus, to our understanding, is the one that fits to the findings from this study. The involvement of this genetic variability in the detection and pathogenesis of BoHV-4 remains to be investigated.Fil: Verna, Andrea Elizabeth. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Área de Investigación en Producción y Sanidad Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Manrique, Julieta Marina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación Playa Unión. Estación de Fotobiología Playa Unión; ArgentinaFil: Pérez, Sandra Elizabeth. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Leunda, M. R.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Área de Investigación en Producción y Sanidad Animal; ArgentinaFil: Pereyra, Susana Beatriz. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Área de Investigación en Producción y Sanidad Animal; ArgentinaFil: Jones, Leandro Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación Playa Unión. Estación de Fotobiología Playa Unión; ArgentinaFil: Odeón, Anselmo Carlos. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Área de Investigación en Producción y Sanidad Animal; Argentin

Effect of Dexamethasone in Neospora Caninum Seropositive Calves

The aim of this study was to evaluate the effect of Dexamethasone (DXM) on clinical parameters, absolute and differential leukocyte count, specific IgG titers, IFN-γ production and parasitemia in beef calves seropositive to Neospora caninum. Fifteen calves were assigned to four experimental groups as follow: group A: 4 sero- positive calves treated with DXM; group B: 4 seropositive calves without DXM; group C: 3 seronegative calves treated with DXM, and group D: 4 seronegative calves without DXM treatment. The absolute leukocyte count was higher in both groups A and C in the 3rd day after the first dose of DXM mainly due to a significant neutrophilia (p0.05). Similarly, IFN-γ levels did not change among experimental groups (p>0.05). DNA was de- tected in calves from groups A and B at 7 Days Post Administra- tion (dpa) and from group A at 14 dpa. Even when DXM adminis- tration induced hemotological parameter changes, no reactivation of Neospora-infection was observed according to specific IgG titers and presence of the parasite on leukocytes in naturally seropositive beef male calves.Fil: Idarraga Bedoya, S.. Universidad Nacional de Mar del Plata; ArgentinaFil: Armendano, Joaquín Ignacio. Universidad Nacional de Mar del Plata; ArgentinaFil: Gual, Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata; ArgentinaFil: Hecker, Yanina Paola. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Leunda, M. R.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Pereyra, S.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Verna, Andrea Elizabeth. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Campero, C. M.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Odeón, Anselmo Carlos. Universidad Nacional de Mar del Plata; ArgentinaFil: Moore, Dadin Prando. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Ice cave reveals environmental forcing of long-term Pyrenean tree line dynamics

1. Tree lines are supposed to react sensitively to the current global change. However, the lack of a long-term (millennial) perspective on tree line shifts in the Pyrenees prevents understanding the underlying ecosystem dynamics and processes. 2. We combine multiproxy palaeoecological analyses (fossil pollen, spores, conifer stomata, plant macrofossils, and ordination) from an outstanding ice cave deposit located in the alpine belt c. 200 m above current tree line (Armeña-A294 Ice Cave, 2, 238 m a.s.l.), to assess for the first time in the Pyrenees, tree line dynamics, and ecosystem resilience to climate changes 5, 700–2, 200 (cal.) years ago. 3. The tree line ecotone was located at the cave altitude from 5, 700 to 4, 650 cal year bp, when vegetation consisted of open Pinus uncinata Ramond ex DC and Betula spp. Woodlands and timberline were very close to the site. Subsequently, tree line slightly raised and timberline reached the ice cave altitude, exceeding its today''s uppermost limit by c. 300–400 m during more than four centuries (4, 650 and 4, 200 cal year bp) at the end of the Holocene Thermal Maximum. After 4, 200 cal year bp, alpine tundra communities dominated by Dryas octopetala L. expanded while tree line descended, most likely as a consequence of the Neoglacial cooling. Prehistoric livestock raising likely reinforced climate cooling impacts at 3, 450–3, 250 cal year bp. Finally, a tree line ecotone developed around the cave that was on its turn replaced by alpine communities during the past 2, 000 years. 4. Synthesis. The long-term Pyrenean tree line ecotone sensitivity suggests that rising temperatures will trigger future P. uncinata and Betula expansions to higher elevations, replacing arctic–alpine plant species. Climate change is causing the rapid melting of the cave ice; rescue investigations would be urgently needed to exploit its unique ecological information

Reconstrucción de la vegetación asociada al depósito tobáceo fluvial holoceno del nogal de el batán, las parras del martín, cordillera ibérica

Development of Quaternary fluvial tufa buildings are often associated with warm and humid phases and are commonly used as climate indicators. El Nogal de El Batán Holocene fluvial tufa buildup, located in Las Parras River valley (Iberian Range, NE Spain), is investigated, following radiocarbon dating, stratigraphic description and palynological analysis. The studied deposit, 8.2 m-thick, is formed of gravels, phytoclast rudstones, calcite- coated stem boundstones and fine carbonate deposits (sands, silts and marls) with peaty horizons and carbonaceous debris. It has been dated within the upper early Holocene (ca. 8400 cal BP) and correlated with the environmental changes reconstructed for the region, identifying at least, three main phases. Pollen results highlight the existence of a conifer landscape, mainy dominated by Pinus nigra/sylvestris type and Juniperus, and locally dense riparian woodland with Corylus, Alnus, Salix, Populus, Celtis, Fraxinus, Tamarix and Rosaceae. Deciduous and evergreen oaks were the main spread regional forests accompanied by many warm-loving taxa like Pistacia, Rhamnus, Arbutus and Olea during the middle Holocene (ca. 8000-5000 cal BP). The transition towards the late Holocene (ca. 4300 cal BP) is evidenced by the rise of herbs and ruderal taxa, and especially the presence of coprophilous fungi, which are linked to the spread of agricultural and grazing practices along the river valley.El desarrollo de edificios tobáceos de origen fluvial se asocia a menudo con fases cálidas y húmedas cuaternarias y son comúnmente empleados como indicadores climáticos. En el presente trabajo se estudia el depósito tobáceo fluvial Holoceno del Nogal de El Batán, situado en el valle del río Las Parras (Cordillera Ibérica, NE España), bajo una perspectiva cronológica, estratigráfica y palinológica. La acumulación tobácea de 8,2 m de espesor, intercala gravas, facies de rudstones de fitoclastos, boundstones de tallos, limos y arenas de composición carbonatada, además de margas, lime mud y niveles turbosos. La acumulación tobácea se emplaza en el Holoceno temprano final (ca. 8400 cal BP) y se correlaciona con los cambios ambientales ocurridos en la región, identificando tres fases principales. Los análisis polínicos señalan un paisaje dominado por coníferas en la base de la secuencia, esencialmente con Pinus nigra/sylvestris tipo y Juniperus, mientras que localmente se desarrolla un bosque ripario donde destacan Corylus, Alnus, Salix, Populus, Celtis, Fraxinus, Tamarix y Rosaceae. En el Holoceno medio (ca. 8000-5000 cal BP) se produce la máxima expansión de las quercíneas, junto con taxones termófilos como Pistacia, Rhamnus, Arbutus y Olea. El incremento de taxones herbáceos y ruderales, y en especial la aparición de esporas coprófilas, marca la expansión agro-pastoril a lo largo del valle fluvial durante el Holoceno tardío (ca. 4300 cal BP)