Comparative Characterization of Enzymatic Digestion from Fish and Soybean Meal from Simulated Digestive Process of Pacific Bluefin Tuna,Thunnus orientalis

The digestive process of the Pacific bluefin tuna (PBT), Thunnus orientalis, was simulated through two phases of in vitro digestion: acidic digestion with porcine pepsin, followed by alkaline digestion with pancreatic crude extract (PCE) obtained from the PBT to hydrolyze fish meal (FM) and soybean meal (SBM) as protein substrates. The crude protein from FM resulted in a lower degree of hydrolysis (73.3%) compared with SBM (79.2%). However, the resulting digested products showed that FM contained 35% more small peptides, with sizes <6.5 kDa than those from the starting material (>150 kDa). The SBM had an increase of only 1.3% in the similar peptide cut‐offs found after hydrolysis. These results suggested that FM appeared to be a better source of protein according to the amount of low‐molecular weight peptides. In addition, the proteolytic activity of PCE showed that 88.9% of its alkaline proteolytic activity corresponded to trypsin and 2.9% corresponded to chymotrypsin activity. The results shown here demonstrate that peptide sizes are important in identifying suitable protein sources for aquafeed production to reinforce the primary results obtained from the in vitro digestibility using the pH‐Stat system. These results also contribute to a better understanding of the digestibility process in aquatic organisms.     

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.     

Anatomical Variations of the Blood Vascular System in Veterinary Medicine. The Internal Iliac Artery of the Dog. Part Two

The aim of this study was to investigate the variability of the internal pudendal artery. Two hundred and thirty‐two pelvic halves from 116 adult dogs were examined. Twenty‐six anatomical variations were found, thirteen occurring in more than 5% of the dogs. Anatomical variations were grouped in relation to the origin of the prostatic/vaginal arteries, middle rectal artery, urethral artery, ventral perineal and caudal rectal arteries. The chi‐squared test was used to analyse differences in sex, side of the body, profile and size, and the results were considered statistically significant when P ≤ 0.05. An identical vascular pattern in both hemipelvises was found for most of the anatomical variations described.     

Anatomical Variations of the Blood Vascular System in Veterinary Medicine: The Internal Iliac Artery of the Dog — Part Three

The aim of this study was to analyse and describe the variability of the umbilical artery. Two hundred and thirty‐two pelvic halves from 116 adult dogs were examined. To study the permeability of the umbilical artery, ten adult dogs, nine newborns and thirteen foetuses between 35 and 50 days of gestation were also used. In relation to the origin of the umbilical artery, six anatomical variations were found. From which five involved a cranial (n = 4) or caudal (n = 1) relocation of its origin, and in one case (n = 1), the umbilical artery arose from the median sacral artery. In eight cases, the umbilical artery gave off the prostatic (n = 1) or vaginal (n = 7) arteries. The permeability of the umbilical artery was the most significant anatomical variation: permeability was detected in 45% (106 of 232 pelvic halves) of all cases, from which 36 were males and 70 females. Interestingly, an equal vascular permeability in both hemipelvises was found for 82% of the dogs, thus additional data related to such feature of the umbilical artery was also recorded. In accordance with the statistical study, the main anatomical variations described showed significant values for gender, side of the body, size and profile variables.     

Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy‐Associated Glycoproteins 1 and 2 in Foetal Fluids

Plasma concentrations of PAG‐1 are used for pregnancy diagnosis and as a marker of placental/foetal well‐being, while those of PAG‐2 may be an indicator of abortion risk in Neospora caninum‐infected cows. Studies have shown that N. caninum infection modifies PAG‐1 and PAG‐2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG‐1, PAG‐2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG‐2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non‐infected foetuses. In infected foetuses, significantly higher concentrations of PAG‐1 (p < 0.001) and PAG‐2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non‐infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG‐1 and PAG‐2 levels and pH values than fluids from non‐infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.     

Gastrointestinal parasitism reduces the plasma availability of doramectin in lambs

A study was undertaken to investigate the effect of parasitism on plasma availability and pharmacokinetic behaviour of doramectin (DRM) in lambs. Fourteen parasitised grey face Suffolk lambs (26.9 ± 1.5 kg bodyweight) were selected for the study. Seven pairs of lambs were allocated to two groups to obtain an approximately even weight distribution. Group I (non-parasitised) was pre-treated with three repeated administrations of 5 mg/kg fenbendazole to maintain a parasite free condition. In group II (parasitised), the lambs did not receive any anthelmintic treatment. After the 85-day pre-treatment period, both groups of animals were treated with DRM by subcutaneous (SC) injection in the shoulder area at 200 μg/kg. Throughout the experimental period, both groups were maintained together under similar feeding and management conditions. Blood samples were collected by jugular venepuncture at different set times between 0.5 h and 60 days post-treatment. After plasma extraction and derivatisation, samples were analysed by high performance liquid chromatography (HPLC) with fluorescence detection. A computerised kinetic analysis was performed and the data were compared using the Student’s paired t test.The parent molecule was detected in plasma between 30 min and either day 20 (parasitised) or day 35 (non-parasitised) post-DRM treatment. The AUC values of the parasitised group (143.0 ± 18 ng d/mL) were significantly lower (P < 0.05) than those observed in the parasitically naïve animals (229.6 ± 21.7 ng d/mL). The mean residence time (MRT) in the parasitised group (3.4 ± 0.3 days) was significantly shorter (P < 0.05) than in the healthy group (6.6 ± 0.6 days). Study results have shown that parasitic disease, through alteration in the body condition, can produce significant changes in the plasma disposition of DRM when administered SC to parasitised lambs.