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

Middle-to-late Holocene palaeoenvironmental reconstruction from the A294 ice-cave record (Central Pyrenees, northern Spain)

Perennial ice deposits in caves represent unique, but underexplored, terrestrial sequences that potentially contain outstanding palaeoclimatic records. Here, we present a pioneer palaeoenvironmental study of an ice deposit preserved in a small sag-type cave (A294) in the Central Pyrenees (northern Iberian Peninsula). The 9.25-m thick sequence, which is dated from 6100 ± 107 to 1888 ± 64 cal BP, represents the oldest known firn ice record worldwide. The stratigraphy (detrital layers, unconformities, and cross stratification), plant macrofossils, and isotopic signature (similarity between the ice linear distribution, δ2H = 7.83δ18O + 8.4, and the Global Meteoric Water Line) of the ice point to the diagenesis of snow introduced to the cave by winter snowstorms. Four phases of rapid ice accumulation (6100–5515, 4945–4250, 3810–3155, and 2450–1890 cal BP) are related to wetter and colder winters. Comparison of the isotopic composition (δ18O and deuterium excess) of the ice with other paleoclimate records show that both source effects and the North Atlantic Oscillation (NAO) mechanism exert a dominant influence on the ice cave record. The NAO signal may be a combination of source effects and rainfall amount. Three intervals with low ice accumulation occurred between the phases of rapid accumulation and were related to drier, and possibly warmer, winters. These centennial-scale episodes appear to be in-phase with regional arid events, as established from high altitude lacustrine records and can be correlated to global Rapid Climate Change events. The current warming trend has dramatically decreased the volume of the ice deposit in cave A294This work was supported by the Spanish Government and the European Regional Development Fund (projects CGL2009-10455/BTE, CTM2013-48639-C2-1-R CGL2016-77479-R). This is a contribution by PaleoQ and Geomorfología y Cambio Global groups (Aragón Government and European Social Fund) and IUCA (University of Zaragoza

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

Retained Austenite Control for the Soft Machining of High-Hardness Tool Steels

Most high-hardness tool steels comprising forming dies require expensive finish machining operations to compensate for the dimensional distortion and surface oxidation caused by the die heat treatment. Precipitation-hardening (PH) tool steels allow for soft finish machining followed by an aging treatment without major deformation or oxidation in the die, but exhibit poor wear performance owing to the lack of carbides in their structure. This drawback can be overcome by combining laser cladding technology, austenite retention, and cryogenic treatments. Hence, an alternative die manufacturing route based on laser cladding was explored. The forming surface of a modified chemistry tool steel die was subjected to cladding. The martensite finish (M-f) temperature of the steel was tuned to enhance austenite retention at room temperature. The cladded surface was then machined in a reduced-hardness condition resulting from retained austenite formation. Subsequent deep cryogenic treatment of the die favoured the retained-austenite-to-martensite transformation, thereby increasing the die hardness without major distortion or oxidation. This process combined the advantages of high-carbide-bearing tool steels and PH steels, allowing for a die with hardness exceeding 58 HRC to be finish machined at <52 HRC. Controlling the occurrence of retained austenite represents an effective strategy for achieving new manufacturing scenarios.The authors gratefully acknowledge the funding provided by the Department of Research and Universities of the Basque Government under Grant No. IT947-16 and the University of the Basque Country UPV/EHU under Program No. UFI 11/29

Seasonal changes in fish assemblages in the River Minho tidal freshwater wetlands, NW of the Iberian Peninsula

The ichthyofauna of the River Minho tidal freshwater wetlands (TFWs) was studied in a semienclosed area, between June 2007 and May 2010, to determine temporal patterns of abundance, biomass and species composition. Fish catches, standardized by the number of fyke nets and by the fishing effort, were analysed and related to river flow, water temperature and precipitation. In total, 21 fish species were identified including six non-indigenous species (NIS) which represented 15% of the total captures (yet 43% of the biomass). Regarding ecological guilds, 82% were freshwater species (61% of the biomass) which included all NIS, whereas 13% of the catches were catadromous species (31% biomass) corresponding to eels. Only 4% were estuarine species (1% biomass) and together marine estuarine opportunists and anadromous species accomplished 1% (2 and 6% biomass, respectively). The fish assemblage showed seasonal patterns with a clear distinction between autumn/winter and spring/summer periods. Significant differences in abundance and biomass were also detected between autumn/winter of 2007/2008 and 2008/2009, i.e., inter-annual variations, with the latter period with lower catches yet higher dominance of NIS. Besides, higher water temperature had a significant negative effect on the observed number of species, but a positive effect on the abundance and biomass of NIS. More attention has to be given to TFWs and their role in structuring fish assemblages, because this information is vital for a sound management, conservation and restoration of estuarine areas.The authors would like to thank Rodrigo Lopez for the study area map arrangement and Eduardo Martins for the field work assistance. This research was partially supported by the European Regional Development Fund (ERDF) through the COMPETE - Operational Competitiveness Programme and national funds through FCT - Foundation for Science and Technology, under the project PEst-/MAR/LA0015/2013

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

GMO Regulatory Aspects of Novel Investigational Vaccine Candidates

Recent scientific and technical developments create novel opportunities for vaccine development. Regulatory compliance has to be ensured from preclinical research to market authorization, whereby different legal frameworks that go beyond quality, efficacy or patient safety aspects need to be taken into account. As academia and start-ups are often focused on gathering scientific evidence, the regulatory maze is often regarded by applicants as challenging in the overall pathway to clinical translation. This is particularly true for applications concerning vaccine candidates containing or consisting of genetically modified organisms (GMOs). Active communication between applicants and competent authorities or advisory bodies early in the development stages facilitates a correct implementation of the regulatory frameworks and is of utmost importance to identify challenges or hurdles in order to avoid unnecessary delay in scientific review. Based on the state-of-play in Belgium, this chapter discusses examples of regulatory journeys of applications with genetically modified viral vectors and novel vaccine candidates that have been reviewed by GMO national competent authorities in Belgium and in Europe. They highlight the need of having a comprehensive view of global perspectives early in the development to facilitate the translation of research to clinical development or even market authorization

Concomitant infection of Neospora caninum and Bovine Herpesvirus type 5 in spontaneous bovine abortions1

Bovine Herpesvirus type 5 (BoHV-5) has not been conclusively demonstrated to cause bovine abortion. Brain lesions produced by Neospora caninum and Bovine Herpesvirus type 1 (BoHV-1) exhibit common features. Therefore, careful microscopic evaluation and additional diagnostic procedures are required to achieve an accurate final etiological diagnosis. The aim of the present work was to investigate the occurrence of infections due to BoHV-1, BoHV-5 and N. caninum in 68 cases of spontaneous bovine abortions which showed microscopic lesions in the fetal central nervous system. This study allowed the identification of 4 (5.9%) fetuses with dual infection by BoHV-5 and N. caninum and 33 (48.5%) cases in which N. caninum was the sole pathogen identified. All cases were negative to BoHV-1. The results of this study provide evidence that dual infection by BoHV-5 and N. caninum occur during pregnancy in cattle; however, the role of BoHV-5 as a primary cause of bovine abortion needs further research. Molecular diagnosis of BoHV-5 and N. caninum confirmed the importance of applying complementary assays to improve the sensitivity of diagnosing bovine abortion.Fil: Marin, Maia Solange. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires. Estación Experimental Agropecuaria Balcarce; Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina;Fil: Morrell, Eleonora Lidia. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires. Estación Experimental Agropecuaria Balcarce; Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina;Fil: Perez, Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Tandil. Centro de Investigacion Veterinaria de Tandil; Argentina;Fil: Leunda, Maria R.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires. Estación Experimental Agropecuaria Balcarce; Argentina;Fil: Moore, Dadin Prando. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires. Estación Experimental Agropecuaria Balcarce; Argentina;Fil: Jones, Leandro Roberto. Fund.Playa Union. Estacion de Fotobiologia Playa Union; Argentina;Fil: Campero, Carlos Manuel. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires. Estación Experimental Agropecuaria Balcarce; Argentina;Fil: Odeon, Anselmo Carlos. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires. Estación Experimental Agropecuaria Balcarce; Argentina

Irlanda: euroa, higiezinen burbuila eta gaur egungo egoera

90. hamarkadaren hasieran Gobernu irlandarrak bertako ekonomian eragin positiboa izango zuten hainbat erabaki estrategiko hartu zituen. Erabaki hauek zergen murrizketak eta ekonomiaren liberalizazioa suposatzen zituzten. Honi esker esportazioak hazkunde ekonomikoaren bultzatzaile nagusi bihurtu ziren. XXI. mendearen hasieran higiezinen sektoreak kanpo sektorea ordezkatuko du ekonomiaren dinamizatzaile nagusi bezala. Euroan sartzeak, atzerriko aurrezkia etxebizitzen erosketara bideratuz, higiezinen burbuilaren sorrera ahalbidetu zuen. Higiezinen sektorean oinarritutako hazkunde ekonomikoa 2007. urtean eztanda egin zuen finantza krisi globalaren ondorioz bukatu egingo da. Gaur egun, Irlandako ekonomia hazkunde jasangaitzeko garai horren ondorioak pairatzen ari da. Hau dela eta, krisi sakoneko egoera bat igarotzen ari da, Europar Batasunak erreskatatu behar izan du eta bere fi nantza sektorearen zati handi bat sektore publikoaren esku dago.KUTXA ; Vicerrectorado de Campus de Gipuzkoa de la UPV/EH

Human–landscape interactions in the Conquezuela–Ambrona Valley (Soria, continental Iberia): From the early Neolithic land use to the origin of the current oak woodland

The sedimentological, geochemical and palynological analyses performed in the Conquezuela palaeolake (41°11′N; 2°33′W; 1124 m a.s.l.) provide a detailed, multiproxy palaeoenvironmental reconstruction in one of the key areas of inner Iberian Neolithic colonization. Combined with archaeobotanical and archaeological data from well-dated settlements along the Conquezuela–Ambrona Valley we investigate how environmental conditions may affect both socio-economic adaptations and livelihood strategies of prehistoric communities. The first evidences of early Neolithic occupation in the valley ca. 7250–6450 cal yr BP (5300–4500 BC) coincided with the onset of a period (7540–6200 cal yr BP, 5590–4250 BC) with higher water availability and warmer climate as alluvial environments were substituted by carbonate-wetland environments in the basin. The Conquezuela record supports an early Neolithic colonization of the inner regions of Iberia favored by warmer and humid climate features and with preferential settlement patterns associated to lakes. The maximum human occupation of the valley occurred during the mid–late Neolithic and Chalcolithic (6200–3200 cal yr BP, 4250–1250 BC) as evidenced by the high number of archaeological sites. Although a number of hydrological oscillations have been detected during this period, the intense landscape transformation at basin-scale, leading to a deforested landscape, was largely a consequence of widespread farming and pastoral practices. Socio-economic activities during Bronze, Iron and Roman times modified this inherited landscape, but the second largest ecosystem transformation only occurred during Mediaeval times when a new agrarian landscape developed with the expansion of stockbreeding transhumance. The current vegetation cover characterized by patches of holm and marcescent oaks and fields reflects an intense human management combining both extensive herding with agrarian activities in order to transform the previous forested landscape into a dehesa-like system.The funding for the present study derives from DINAMO2 (CGL-BOS 2012-33063) and AGRIWESTMED (ERC Grant Agreement #230561) projects, provided by the Spanish Inter-Ministry Commission of Science and Technology (CICYT) and the European Research Council under the European Union's Seventh Framework Programme (FP7/2007–2013). XRF data were obtained at the XRF Core Scanner Laboratory (CRG Marine Geosciences, University of Barcelona). Josu Aranbarri acknowledges the predoctoral funding provided by the Basque Country Government (ref: FI-2010-5). Graciela Gil-Romera hold a post-doctoral contract funded by “Juan de la Cierva” (ref: JCI2009-04345) program. Eduardo García-Prieto and Maria Leunda are supported by predoctoral FPI grants BES-2010-038593 and BES-2013-063753, respectively. We also thank Elena Royo for her help with the lab procedures and the two anonymous referees for their valuable suggestions.Peer reviewe