Effect of wilting,silage additive,PEG treatment and tannin content on the distribution of N between different fractions after ensiling of three different sainfoin (Onobrychis viciifolia) varieties

Sainfoin (Onobrychis viciifolia) is a tanniniferous, leguminous plant that has potentially beneficial effects on protein utilization in ruminants. As ensiling causes protein breakdown and elevated levels of buffer soluble N (BSN), we studied the distribution of N before and after ensiling sainfoin. Three varieties of sainfoin were either direct‐cut and frozen directly or wilted and frozen before later ensiling in mini‐silos with and without acidification with Promyr (PM; an acidifying commercial mixture of propionic and formic acid) and with or without polyethylene glycol (PEG). Extractable tannins (ET) and protein‐bound tannins (PBT) were measured with an HCl/butanol method in an attempt to correlate tannin levels to N fractions. The sainfoin silages showed good ensiling characteristics and had relatively high concentrations of undegraded protein. The effect of wilting on BSN levels (g/kg N) was dependent on sainfoin variety (P < 0·001). PEG increased and PM decreased the level of BSN in the silages (P < 0·001). PM treatment also produced less non‐protein N and ammonia‐N (P < 0·05) as compared with no additive. Addition of PEG to the silage increased the BSN‐proportion 1·8‐ and 2·6‐fold for both DM stages. A strong tannin‐protein binding effect is, therefore, confirmed in sainfoin. However, correlations between tannin levels (ET and PBT) and BSN were poor in the (non‐PEG) silages, indicating either that the HCl/butanol method is unsuitable for measuring tannin in silages or that qualitative attributes of tannins are more relevant than quantitative. The HCl/butanol method seems therefore not to be useful to predict degradation of protein in sainfoin silages.     

Comparative pharmacokinetics and pharmacokinetic/pharmacodynamic analysis by nonlinear mixed‐effects modeling of cefquinome in nonpregnant,pregnant, and lactating goats after intravenous and intramuscular administration

Cefquinome is a fourth‐generation cephalosporin that is used empirically in goats. Different physiologic factors like pregnancy or lactation could determine the pharmacokinetic behavior of drugs in the organism. The objectives of this study are to (a) compare the pharmacokinetics of cefquinome after intravenous and intramuscular administration in adult nonpregnant (n = 6), pregnant (n = 6), and lactating goats (n = 6), at a dose of 2 mg/kg, with rich sampling by nonlinear mixed‐effects modeling, (b) conduct a pharmacokinetic/pharmacodynamic analysis to evaluate the efficacy of the recommended posology in goats with different physiological states, and (c) determine the optimal posology that achieve a PTA value ≥ 90%, taking into account a T > MIC ≥ 60% of a MIC value ≤ 0.25 µg/ml, in the different subpopulations of goats for both routes. Gestation significantly increased Ka and V1, while reduced F0, Cl, and Q. On the other hand, lactation significantly increased V1 and reduced Tk0. Cefquinome concentrations achieved in placental cotyledon, amniotic fluid, and fetal serum indicate a minimal penetration across the placental barrier. Moreover, milk penetration of cefquinome was minimal. The total body clearance of cefquinome for goats was 0.29 L kg^?1 hr^?1, that is apparently higher than the reported for cows (0.13 L kg^?1 hr^?1) and pigs (0.16 L kg^?1 hr^?1). So, the optimal dose regimen for cefquinome after intravenous and intramuscular administration required higher dose and frequency of administration compared with recommendations for cows or pigs. Therefore, 2 mg kg^?1 8 hr^?1 and 5 mg kg^?1 12 hr^?1 could be used for IV and IM routes, respectively, for the treatment of respiratory infections caused by P. multocida and M. haemolytica, but only 5 mg kg^?1 12 hr^?1 by both routes should be recommended for Escherichia coli infections.     

Swine viscera preservation in hypersaturated salt solution after alcohol fixation as a preparation method for educational purposes

The use of live animals for educational purposes is an old practice that is still employed in teaching and research institutions. However, there are several objections to this practice, whether for ethical or humanitarian reasons. Surgical techniques teaching using anatomical pieces and/or preserved cadavers promotes greater learning efficiency, provides exercise repetition and increases the confidence and satisfaction of the students when compared to the use of live animals. The current work aimed to analyse the feasibility of using fresh swine urinary bladder and small intestines (jejunum), obtained from slaughterhouses, fixed in 99.8% ethyl alcohol (EA) and preserved in sodium chloride hypersaturated solution (SCHS) at 30%, for 7, 14 and 21 days, as an alternative method for surgical skills training (SST). Swine viscera, fixed in EA and preserved in SCHS, presented a realistic appearance, absence of odour and maintained the viable morphological characteristics during the performance of the operative techniques. Preservation solutions had low cost, were easy to acquire and did not offers risks to human health. Therefore, urinary bladders and small intestines fixed in 99.8% EA for 30 days and maintained in 30% SCHS at different periods were demonstrated as a good viable option as a preservation method for surgical skills training.     

Optimizing injection time of GFP plasmid into sheep zygote

Microinjection of exogenous DNA into the cytoplasm of matured oocytes or zygotes is a promising technique to generate transgenic animals. However, the data about the microinjection time and procedure in sheep are limited and have not treated in detail. To obtain more in-depth information, the Sarda sheep oocytes from abattoir-derived ovaries were subjected to IVM and IVF. Then, the GFP plasmid as a reporter gene was injected into the cytoplasm of MII oocytes (n: 95) and zygotes at different post-insemination intervals (6–8 hpi, n: 120; 8–10 hpi, n: 122; 10–12 hpi, n: 110 and 12–14 hpi, n: 96). There were no significant differences in the cleavage rates between the groups. However, blastocyst rate of injected zygotes at all-time intervals was significantly lower than injected MII oocytes and control group (p < 0.05). Interestingly, the proportion of GFP-positive embryos was higher at 8–10 hpi compared with other injected groups (4 % versus 0 %, p  < 0.01). Among these, the proportion of mosaic embryos was high and two of those embryos developed to the blastocyst stage. In conclusion, we settled on the cytoplasmic microinjection of GFP plasmid at 8–10 hpi as an optimized time point for the production of transgenic sheep and subsequent experiments.     

Use of dimethylformamide to cryopreserve alpaca semen previously incubated with collagenase

The objective of this study was to evaluate the effect of collagenase and two final dimethylformamide (DMF) concentrations (4% and 7%) on alpaca frozen-thawed sperm quality. A total of 25 ejaculates from 5 alpaca were obtained using electroejaculation. Each individual ejaculate was evaluated and then diluted 4:1 in a solution of 1 mg/ml collagenase in HEPES-TALP medium and incubated for 4 min at 37°C. Subsequently, samples were diluted in TRIS-fructose-citric acid-egg yolk and cooled to 5°C. Then, each sample was divided in two aliquots and DMF at final concentration of 4% or 7% was added, equilibrated for 1 hr at 5°C and frozen over liquid nitrogen vapours. A Kruskal–Wallis test was used to evaluate the sperm morphometry, and Completely Random Block designs were used to analyse sperm motility, viability, membrane function and acrosome status. After collagenase incubation, none of the samples showed thread formation, and sperm parameters were preserved. Non-progressive motile sperm were higher (p < .05) in equilibrated samples (4% DMF: 31.8 ± 8.3% and 7% DMF: 36.3 ± 11.8%) compared to raw (10.1 ± 4.3%) and frozen-thawed semen (4% DMF: 9.7 ± 1.8% and 7% DMF: 7.5 ± 3.2%). Sperm membrane function, membrane integrity and intact acrosomes were higher (p < .05) in raw semen (40.1 ± 12.2%, 94.6 ± 3.2% and 91.3 ± 8.1%) compared to frozen-thawed samples (4% DMF: 19.8 ± 4.7%, 53.2 ± 2.7%, 65.7 ± 8.7% and 7% DMF: 20.4 ± 4.5%, 54.1 ± 1.4%, 64.6 ± 9.1%). Length of the sperm head was lower in frozen-thawed samples, being statistically different with 4% DMF compared to pre-freezing samples. The ratio between acrosome and head areas was greater (p < .05) in frozen-thawed samples. Incubation of raw alpaca semen with collagenase decreased the thread formation without affecting sperm quality. Frozen of collagenase treated alpaca semen with 4% or 7% DMF did not preserve the sperm parameters in thawed samples.     

Impact of Land Use and Land Cover Changes on Organic Carbon Stocks in Mediterranean Soils (1956–2007)

During the last few decades, land use changes have largely affected the global warming process through emissions of CO₂. However, C sequestration in terrestrial ecosystems could contribute to the decrease of atmospheric CO₂ rates. Although Mediterranean areas show a high potential for C sequestration, only a few studies have been carried out in these systems. In this study, we propose a methodology to assess the impact of land use and land cover change dynamics on soil organic C stocks at different depths. Soil C sequestration rates are provided for different land cover changes and soil types in Andalusia (southern Spain). Our research is based on the analysis of detailed soil databases containing data from 1357 soil profiles, the Soil Map of Andalusia and the Land Use and Land Cover Map of Andalusia. Land use and land cover changes between 1956 and 2007 implied soil organic C losses in all soil groups, resulting in a total loss of 16·8 Tg (approximately 0·33 Tg y⁻¹). Afforestation increased soil organic C mostly in the topsoil, and forest contributed to sequestration of 8·62 Mg ha⁻¹ of soil organic C (25·4 per cent). Deforestation processes implied important C losses, particularly in Cambisols, Luvisols and Vertisols. The information generated in this study will be a useful basis for designing management strategies for stabilizing the increasing atmospheric CO₂ concentrations by preservation of C stocks and C sequestration. Copyright © 2012 John Wiley & Sons, Ltd.     

Effects of Annealing and Concentration of Calcium Salts on Thermal and Rheological Properties of Maize Starch During an Ecological Nixtamalization Process

The objective of the present work was to study the effect of annealing and concentration of Ca(OH)₂ (lime) and calcium salts on the thermal and rheological properties of maize starch during an ecological nixtamalization process. Thermal and rheological properties of maize starch changed during the ecological nixtamalization process because of three main causes: the annealing phenomenon, type of calcium salt, and calcium salt concentration. In all treatments thermal properties (T_o, T_p, and T_f) of nixtamal starch increased owing to the annealing process, whereas the type of salt or lime increased thermal properties and decreased pasting properties in this order: CaCl₂ > CaSO₄ > Ca(OH)₂ ≈ CaCO₃. This behavior was because of the dissociation of each salt or lime in water. Anions (OH⁻) can penetrate much more easily into the starch granule and start the gelatinization process by rupturing hydrogen bonds. Additionally, amylose‐lipid complexes were formed during the nixtamalization processes, as indicated by an increasing peak at 4.5 Å in X‐ray diffraction patterns.