The efficiency of in vitro ovine embryo production using an undefined or a defined maturation medium is determined by the source of the oocyte

In vitro oocyte maturation can be influenced by oocyte source and maturation media composition. The aim of the present study was to compare the efficiency of a defined in vitro maturation medium (TCM199 supplemented with cysteamine and epidermal growth factor; Cys + EGF) with an undefined medium (TCM199 supplemented with follicle-stimulating hormone and follicular fluid; FSH + FF) for in vitro production (IVP) of ovine embryos, using oocytes obtained by laparoscopic ovum pick-up from FSH-stimulated [n=11; 158 cumulus-oocyte complexes (COCs)] and non-stimulated (n=16; 120 COCs) live ewes, as well as abattoir-derived oocytes (170 COCs). The produced blastocysts were vitrified and some of them were transferred to synchronized recipients. The best and the worst final yields of embryo IVP observed in this study were obtained using oocytes from FSH-stimulated ewes matured in FSH + FF (41.3%; 33/80) and in Cys + EGF (19.2%; 15/78) medium, respectively (p<0.01). No significant differences between both media were attained in the blastocyst development rate or in the final yield of embryo IVP using oocytes from non-stimulated ewes or abattoir-derived oocytes. The overall in vivo survival rate of the transferred vitrified blastocysts was 13.1% (8/61), without significant differences between oocyte sources or maturation media. In conclusion, under the experimental conditions of the present study, TCM199 supplemented with cysteamine and EGF is a convenient defined maturation medium for IVP of embryos from oocytes of live non-stimulated ewes or from oocytes of abattoir-derived ovaries. However, the best final yield of embryo IVP observed in this study was attained when oocytes came from FSH-stimulated donors and TCM199 was supplemented with FSH and follicular fluid.     

Subunit influenza vaccine candidate based on CD154 fused to HAH5 increases the antibody titers and cellular immune response in chickens

World Health Organization has a great concern about the spreading of avian influenza virus H5N1. To counteract its massive spread, poultry vaccination is highly recommended together with biosecurity measures. In our study, a recombinant vaccine candidate based on the fusion of extracellular segments of hemagglutinin (HA) H5 of avian influenza virus and chicken CD154 (HACD) is tested with the aim of enhancing humoral and cellular immune responses in chickens. Protein expression was carried out by transducing several mammalian cell lines with recombinant adenoviral vectors. HACD purification was assessed by three distinct purification protocols: immunoaffinity chromatography by elution at acidic pH or with a chaotropic agent and size exclusion chromatography. Humoral and cellular immune responses were measured using the hemagglutination inhibition assay and the semiquantitative real time PCR, respectively. The results showed that humoral response against HACD was significantly higher than the obtained with HA alone after booster (P<0.01, P<0.05). From HACD molecules purified by distinct protocols, only the obtained by size exclusion chromatography generated hemagglutinationin-inhibition activity. IFN-γ levels indicated that cellular immune response was significantly higher with HACD, in its pure or impure form, compared to its counterpart HA (P<0.01). These data demonstrate that HACD is able to significantly enhance humoral and cellular immune responses against HA antigen, which make this fusion protein a promising subunit vaccine candidate against H5N1 virus outbreaks.     

New techniques for an old disease: sarcoptic mange in the Iberian wolf

Sarcoptic mange, a parasitic skin infection caused by the burrowing mite Sarcoptes scabiei, has been reported in over 100 mammals, including humans. In endangered species, mange causes conservation concerns because it may decimate isolated populations and contribute to extinction. The Iberian Peninsula still maintains one of the largest wolf (Canis lupus) populations in Europe. In Iberia, sarcoptic mange is endemic in red foxes (Vulpes vulpes) and the first confirmed wolf mange cases were recently reported. However, knowledge on S. scabiei in wolves is scarce because of the sampling difficulties inherent to research on scarce species. In order to describe wolf mange epidemiology and to infer conservation implications, this study combined traditional laboratory techniques with the revision of wolf carcass pictures taken by field biologists and original information obtained by camera trapping. A total of 125 necropsies and 8783 camera-trap days allowed insights into wolf mange epidemiology between 2003 and 2010. Living Sarcoptes mites were detected in 19% of the fresh carcasses. Alopecic (delayed) type IV hypersensitive response reactions were observed, while parakeratotic lesions were infrequent. The number of mites isolated per wolf ranged from 1 to 78, and had a negative correlation with the percentage of alopecic skin. No effect by sex on mange prevalence was found. Yearlings showed a lower probability to present mange-compatible lesions than pups or adults. Wolves with mange-compatible lesions had a lower kidney fat index than apparently healthy ones. ELISA testing of 88 sera yielded an antibody prevalence of 20%. Photo-trapping recorded mange-compatible lesions since 2003 with a peak in 2008. The percentage of wolves with mange-compatible lesions registered in camera-traps during 1 year correlated with the percentage of red foxes with lesions in the previous year. This is the first large survey on sarcoptic mange in the Iberian wolf. Necropsy data, with alopecia as the main feature and a slight effect on body condition, and trends derived from camera trapping coincided in showing a rather low prevalence and an apparently stable situation of the disease and its host, suggesting that this parasite is currently not a major threat for this wolf population. However, more information is needed in order to assess the effect of mange on aspects such as pup survival.     

Assessing the potential effect of different land cover management practices on sediment yield from an abandoned farmland catchment in the Spanish Pyrenees

Purpose  

Sediment delivery from headwater catchments to reservoirs is a serious threat to reservoir sustainability and is a critical issue in Mediterranean environments where water resources are scarce. In this study we assessed the consequences of two landscape management scenarios (natural vegetation recovery and scrub clearance) on soil erosion and sediment yield. The results were analyzed in relation to predicted and measured rates of soil erosion and sediment yield, with the aim of promoting better management practices.     

Pigs naturally exposed to porcine circovirus type 2 (PCV2) generate antibody responses capable to neutralise PCV2 isolates of different genotypes and geographic origins

Porcine circovirus type 2 (PCV2) is the essential infectious agent for PCV2-systemic disease (PCV2-SD, formerly known as postweaning multisystemic wasting syndrome) and other pathological conditions. Recent studies indicated antigenic variability amongst different PCV2 isolates and suggested that single amino acid changes within the capsid protein determine differences in the level of neutralization by specific monoclonal antibodies. The objective of the present study was to examine the cross-reactivity of PCV2 antibodies induced in the context of a natural infection against different PCV2 isolates belonging to genotypes PCV2a and PCV2b. Sera taken from several farms from animals of varying health status (PCV2-SD and age-matched healthy pigs and a set of slaughter-aged animals) were assayed for neutralizing activity against four PCV2 isolates from both predominant genotypes (PCV2a and PCV2b) and of differing geographic origins (Europe and North-America). Results showed that most of studied pigs (79 out of 82) contained neutralizing antibodies (NA) able to neutralize all four studied viral strains. Overall, pigs had significantly higher NA titres against PCV2a than against PCV2b (P < 0.001). Accordingly, studied serums were able to better neutralize Burgos390L4 and Stoon-1010 strains (PCV2a) than L-33-Sp-10-54 and MO/S-06 strains (PCV2b) (P < 0.001). No differences between capabilities of seroneutralization of viruses from different geographic origin were observed. Present data suggests that sequence differences between PCV2 isolates translate to functional antigenic differences in viral neutralization in vivo.     

Humus accumulation, humification, and humic acid composition in soils of two post-mining chronosequences after coal mining

Purpose

To assess how the rates of humus formation and humification are affected by land use and age of ecosystems, this study investigated soil development in two post-mining chronosequences (spoil heaps formed from open-cast coal mining near Sokolov, Czech Republic). The following characteristics were measured: content, composition, and properties of humic acids; organic carbon (C) and total nitrogen (N) contents; pH; and amorphous iron content.

Materials and methods

Two chronosequences were studied. One consisted of unreclaimed spontaneously revegetated spoils (3, 12, 20, and 40 years old). The other consisted of heaps that were reclaimed by planting alder in graded heaps (7, 15, 20, 30, and 40 years old). Humus and iron contents as well as pH were determined, and humic acids were extracted for detailed chemical analyses (C, H, N, ¹³C-NMR, and pyrolysis mass-spectrometry).

Results and discussion

C and N accumulated faster in the reclaimed sites than in the unreclaimed sites; organic matter accumulation results in the decrease of pH in all soils and in spontaneous sites also increase of amorphous iron content. Humic acids (HA) and fulvic acids (FA) increased with the site age in both chronosequences. The CHA/CFA was higher in the reclaimed soils than in the unreclaimed soils, and the CHA/CFA ratio increased with age in the unreclaimed soils. Humic acid aromaticity was higher in the reclaimed site than in the unreclaimed sites but increased with the age of unreclaimed sites. H and O content decreased with age of reclaimed soils. Humification led to an increase in HA caloricity in the reclaimed sites but not in unreclaimed sites because of the influence of residual wax-type substances in the unreclaimed sites. Degree of HA oxidation (ω) for the reclaimed plots was similar to that in typical zonal soils but was highly variable in unreclaimed soils.

Conclusions

Relative to spontaneous revegetation in the unreclaimed sites, reclamation increased the rates of humus accumulation, humification, and humus acid transformations. The differences between reclaimed and unreclaimed sites, however, decreased with site age and were very small in 40-year-old sites. These differences correspond to the rapid colonization of the reclaimed sites and the slow colonization of the unreclaimed sites by soil biota.     

Blood mineral profile of the critically endangered Western Derby eland (Taurotragus derbianus derbianus) in two conservation breeding reserves in Senegal

For many nondomestic species, nutritional requirements and the challenges faced in their current habitats are unknown. This is the case of small semi-captive population of the critically endangered Western Derby eland held in two wildlife reserves (the Bandia and the Fathala) in Senegal. The aim of this study was to determine the mineral profile (Ca, P, Mg, Cu, Fe, Zn, Se, K, S) in blood serum and to identify potential mineral deficiencies. Serum data (11 individuals) were compared to other Tragelaphineae, where it was lower in almost all elements. Considerably low concentrations were recorded for Cu, Fe and Zn. Animals in the Bandia reserve had higher serum levels of Fe and K compared to the Fathala reserve and a higher serum level of S in the Fathala reserve compared to the Bandia reserve. Recorded mineral levels may reflect most likely the limited mineral background in the local environment. The knowledge of adequate nutritional requirements and health status of these animals is relevant for the conservation breeding programme. No other serum mineral reference values exist for Western Derby eland, neither for free-ranging nor captive animals. Therefore, the knowledge of reference intervals for minerals in serum may serve for monitoring of the population's health.     

Connections between the internal and the external capsules and the globus pallidus in the sheep: A dichromate stain X-ray microtomographic study

Sheep are recognized as useful species for translational neurodegeneration research, in particular for the study of Huntington disease. There is a lack of information regarding the detailed anatomy and connections of the basal ganglia of sheep, in normal myeloarchitectonics and in tract-tracing studies. In this work, the organization of the corticostriatal projections at the level of the putamen and globus pallidus (GP) are explored. For the first time, the myeloarchitectonic pattern of connections between the internal (IC) and the external (EC) capsules with the GP have been investigated in the sheep. Formaldehyde-fixed blocks of the striatum were treated with a metallic stain containing potassium dichromate and visualized using micro-CT (µ-CT). The trivalent chromium (Cr3+), attached to myelin phospholipids, imparts a differential contrast to the grey and white matter compartments, which allows the visualization of myelinated fascicles in µ-CT images. The fascicles were classified according to their topographical location in dorsal supreme fascicles (X, Y, apex) arising from the IC and EC; pre-commissurally, basal fascicles connecting the ventral part of the EC with the lateral zone of the ventral pallidum (VP) and, post-commissurally, superior (Z₁), middle (Z₂) and lower (Z₃) fascicles, connecting at different levels the EC with the GP. The results suggest that the presumptive cortical efferent and afferent fibres to the pallidum could be organized according to a dorsal to ventrolateral topography in the sheep, similar to that seen in other mammals. The proposed methodology has the potential to delineate the myeloarchitectonic patterns of nervous systems and tracts.