Digestive Enzyme Activity during Larval Development of Black Snook,Centropomus nigrescens

Black snook, Centropomus nigrescens, have been identified as a promising candidate for aquaculture although, like many of the Centropomid species, high mortality associated with early larval stages presents a significant bottleneck to their commercialization. The digestive capacity of black snook larvae throughout the first 37 d after hatch (d.a.h.) was evaluated by quantifying digestive enzyme activities using biochemical techniques. Results showed that black snook larvae have alkaline proteases at hatching, which are known to be important during the first days of feeding for digestion. Toward the end of the study, acid proteases concentration increased (37 d.a.h.). Enzymes for lipid digestion, pancreatic lipase and bile salt‐activated lipase, were already present in the larvae before exogenous feeding commenced, and their activity increased with age and growth (length). Intracellular digestion, measured as the activity of leucine‐alanine peptidase, was high early on (5 d.a.h.) and decreased as development progressed (next 32 d). In contrast, alkaline phosphatase activity was lowest at first feeding and subsequently increased with age. Overall patterns in enzyme activity suggest the possibility of live feed weaning before 32 d.a.h. if artificial diets can be properly balanced.     

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.     

Early interspecific dynamics,dry matter production and nitrogen use in Kernza (Thinopyrum intermedium) – alfalfa (Medicago sativa L.) mixed intercropping

ABSTRACT

The global interest in growing perennial grain crops such as intermediate wheatgrass (Thinopyrum intermedium) (Kernza) for production of food and feed is increasing. Intercropping Kernza with legumes may be a sustainable way of supplying nitrogen to soil and associated intercrop. We determined the competitive interactions between intercropped Kernza (K) and alfalfa (Medicago sativa L.) (A) under three inorganic nitrogen (N) rates N0, N1, N2 (0, 200, 400 kg ha^?1) and five species relative frequencies (SRF) (100%K:0%A, 75%K:25%A, 50%K:50%A, 25%K:75%A and 0% K:100%A) in mixed intercrops (MI) in a greenhouse pot experiment. After 11 weeks of growth. Kernza dry matter yield (DM) and N accumulated (NACC) were low, but alfalfa DM and NACC high at 0 kg N ha^?1. 200 and 400 kg N ha^?1 fertiliser application increased the competitive ability (CA) of Kernza and reduced the CA of alfalfa. SRF had large impacts on alfalfa DM, NACC and NFIX only at 0 kg N ha^?1 fertiliser, and insignificant impacts on Kernza at all N fertiliser levels, indicating that adjustment of SRF may not be an effective way to modulate the interspecific competition of Kernza. Further research on the other factors that influence the interspecific competition are warranted.     

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.     

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.     

Venom immunotherapy for Hymenoptera allergy in a dog

A 1.5-year-old male castrated dog was presented in anaphylactic shock after suffering an apparent bee sting. Immunotherapy with bee venom was initiated based upon history, skin testing and serological testing for allergen-specific immunoglobulin (Ig)E. The dog was maintained on venom immunotherapy for five years and showed no signs of adverse effects from therapy or from repeated bee stings.     

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.