Renal handling of calcium and phosphorus in experimental renal hyperparathyroidism in dogs

Twenty-four hour urinary excretion, fractional excretion and the filtered load of calcium and phosphorus were monitored as hyperparathyroidism evolved in a model of progressive canine renal failure. Thirteen beagles of both sexes aged four and a half months were used. Nine of them were subjected to a renal damaging schedule (neomycine, 60 mg/kg/48 h, IM, 32 weeks) in order to induce chronic renal failure leading to secondary hyperparathyroidism (2HPT group). The remaining four were kept as the control group. The experiment was conducted over 32 weeks. Blood and 24 h urine were collected every four weeks. Calcium, phosphorus and creatinine were analyzed. Plasma parathormone and calcitonin were determined at weeks 0, 12, 24 and 32. The level of renal function in the 2HPT animals was reduced to 25% of that of the controls (endogenous creatinine clearance was 0.45 +/- 0.22 mL/min/kg as opposed to 1.81 +/- 0.54 mL/min/kg). Hyperparathyroidism was confirmed by a progressive increase in the levels of the parathyroid hormone. Calcitonin levels were not modified. A tendency to hypocalcaemia was observed, reaching statistically significant levels from the twenty-eighth week of the study, when hyperphosphataemia also became significant. Daily urinary excretion of calcium and phosphorus remained at values considered normal throughout the experiment with no alteration imputable to the impaired renal function. This is explained by the decrease in the filtered load of these elements (in both cases statistically significant from the 24th week on) being associated with an increase in their fractional excretion. Thus, calcium and phosphorus urinary excretion values could be maintained in a normal range up to the end of the experiment, showing that renal calcium handling in dogs with experimentally induced renal failure seems to differ from that observed in human patients.     

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.     

Extracellular proteins of Clostridium chauvoei are protective in a mouse model

The anaerobic bacillus Clostridium chauvoei is the causative agent of blackleg, a lethal disease that has an important impact on the sheep and cattle industry worldwide. Immunity to C. chauvoei is considered to be mainly anticellular, and for this reason there is scarce information about the immunogenicity of extracellular proteins. In this work variations in protein profiles, immune response by ELISA and protective capacity of culture supernatants of three C. chauvoei strains, collected at different growth phases, are reported. Sera raised against extracellular antigens also recognised cellular antigens of the same molecular masses. Partially purified cell-free supernatants and those concentrated 10 times by ultrafiltration (C-CFS), obtained at the early stationary phase of growth, induced a strong immunoprotective response, even at low doses, that was more marked for C. chauvoei strain ATCC 10092 (p < or = 0.05). With C-CFS formulations, a clear relationship was observed between IgG titres, protective capacity and concentration of the antigen doses, indicating a specific immune response.     

Effects of dietary arginine on hematological parameters and innate immune function of channel catfish

The effects of elevated dietary arginine on the hematology and immune function of juvenile channel catfish Ictalurus punctatus were evaluated by means of in vivo and in vitro experiments. Healthy juvenile channel catfish (average weight, 34.8 g) were fed casein-gelatin-based diets containing 28% crude protein and supplemented with crystalline L-arginine (ARG) at 0.5, 1, 2, or 4% of diet. An intact-protein diet containing 1.3% arginine also was included to investigate the effects of amino acid form (crystalline-free amino acids versus intact protein). Each purified diet was fed to apparent satiation to triplicate groups of fish for 6 weeks. At the end of the experimental feeding period, the fish were injected intraperitoneally with two doses (3 d apart) of 2 mg lipopolysaccharide/kg body weight. Six days after the initial injection, the fish were anesthetized and tissue samples were obtained to evaluate hematological and humoral and cellular immune parameters, including phagocytic activity of peritoneal macrophages, hemoglobin, hematocrit, mean corpuscular volume (MCV), blood cell counts, plasma protein, and hepatic superoxide dismutase activity. High dietary levels (4% ARG) resulted in significantly higher levels of hemoglobin, hematocrit, and circulating erythrocytes. Dietary ARG did not significantly affect MCV and the number of circulating leukocytes, lymphocytes, eosinophils, and monocytes. In vitro, a moderate level (2 mM) of ARG in the culture media was found to be ideal in significantly enhancing phagocytosis. This study demonstrates that some aspects of the immune system of channel catfish are sensitive to changes in dietary ARG.     

Determination of in vitro digestibility of forage species used in ruminant feeding

Within the evaluation of the quality of forage resources, the main parameter that defines it is the digestibility of dry matter, which together with the amount of neutral and acidic detergent fibers and crude protein constitutes the basic information to assess forages which are supplied in the diet of the cattle. This research was carried out at the University of Los Llanos (Villavicencio, Colombia), and its objective was to determine the digestibility of three forages in cattle through three different in vitro techniques: inoculation with ruminal fluid and with feces and enzymatic digestibility technique, making the comparison with the in situ technique in order to validate the techniques and equipment that are being used for these procedures. The following species were evaluated: Pennisetum purpureum (PP), Tithonia diversifolia (TD), and Bauhinia variegata (BV), assessing the curve and rate of degradation of dry matter (DM), neutral detergent fiber (NDF), and total protein (TP) (0 to 72 h). A design of repeated measures was used, under which the analysis of variance was carried out to determine the ranges of deviation between the techniques and thus establish the trend of the data; the variables evaluated were the DM, NDF, and TP digestibilities of the three forages using the four techniques (three in vitro and one in situ). After verifying the differences between the variances of the digestibilities and checking the sphericity assumption with the Mauchly test, multiple comparisons were made with the Bonferroni test with a significance of 5%. The digestibility of DM, NDF, and TP varied between 38.62 and 44.22, 54.18 and 66.97, and 47.54 and 57.05%; 49.07 and 70.70, 72.52 and 75.44, and 62.61 and 74.02%; 29.93 and 34.84, 26.21 and 70.88, and 25.67 and 50.60% respectively in forages PP, TD, and BV, depending on the technique used for their estimation. Despite finding statistically significant differences between several of the comparisons made in the digestibility techniques, a high coefficient of determination and a high correlation between the in vitro estimations with respect to the in situ estimation were found; therefore, it is possible to use these techniques routinely thus avoiding the need to have cattle with fistulae to perform digestibility tests, with enzymatic digestibility technique being the most practical one.

     

Use of sewage sludge in silvopastoral systems under Pinus radiata D. Don: soil,tree growth,and pasture production

Agroforestry Systems - Short-term production in silvopastoral systems is often limited due to inappropriate soil-fertility management. Sewage sludge fertilisation could enhance productivity of...     

Do pine plantations provide mycorrhizal inocula for seedlings establishment in grasslands from Patagonia,Argentina?

We investigated if Pinus ponderosa plantations in Patagonia are able to produce viable mycorrhizal inocula towards adjacent grasslands, which only harbor endomycorrhizal vegetation. We hypothesized that these inocula have the potential to contribute to the establishment of naturally disseminated seedlings. Also, we determined the main fungal taxa involved in this process. Seven plantations in the onset of their reproductive phase and located in the Patagonian native forest/steppe ecotone (Argentina) were selected. Soil samplings were obtained at nine points along a 450 m long, W-E transect established in each plantation. Soil bioassays were performed in a greenhouse, with P. ponderosa seedlings acting as hosts for mycorrhizal inocula present in soil samples, during 12 months. Mycorrhization percentage, morphotype richness and morphotype composition was determined through morphological evaluation. Viable ecto- and ectendomycorrhizal inocula were found disseminated outside plantations. The amount of mycorrhizal inoculum followed a decreasing function with distance to plantation edges. Mycorrhizal fungal genus Rhizopogon and “E-strain” mycorrhizal types appeared as pioneering taxa regarding seedlings colonization, being the most persistent and frequent symbionts found. Plantations, thus, facilitate the surrounding terrain for newcoming seedlings through the dispersion of mycorrhizal fungal inocula.