Agarose gel electrophoresis of cerebrospinal fluid proteins of dogs after sample concentration using a membrane microconcentrator technique

BACKGROUND: Cerebrospinal fluid (CSF) is produced in the cerebral ventricles through ultrafiltration of plasma and active transport mechanisms. Evaluation of proteins in CSF may provide important information about the production of immunoglobulins within the central nervous system as well as possible disturbances in the blood-brain barrier. OBJECTIVE: The objective of this study was to measure the concentration and fractions of protein in CSF samples using a membrane microconcentrator technique followed by electrophoresis, and to compare the protein fractions obtained with those in serum. METHODS: CSF samples from 3 healthy dogs and 3 dogs with canine distemper virus infection were concentrated using a membrane microconcentrator having a 0.5 to 30,000 d nominal molecular weight limit (Ultrafree, Millipore, Billerica, MA, USA). Protein concentration was determined before and after concentration. Agarose gel electrophoresis was done on concentrated CSF samples, serum, and serial dilutions of one of the CSF samples. RESULTS: Electrophoretic bands were clearly identified in densitometer tracings in CSF samples with protein concentrations as low as 1.3 g/dL. The higher CSF protein concentration in dogs with distemper was mainly the result of increased albumin concentration. CONCLUSION: The microconcentrating method used in this study enables characterization of the main protein fractions in CSF by routine electrophoresis and may be useful for interpreting the underlying cause of changes in CSF protein concentrations.     

Haematologic and immunologic parameters of bullfrogs,Lithobates catesbeianus,fed probiotics

The effects of two probiotics (P₁–Lactobacillus acidophilus, Bifidobacterium bifidum and Enterococcus faecium and P₂–Bacillus subtilis) supplemented to commercial feed (40% crude protein) on the haematological and immunological parameters of the bullfrog Lithobates catesbeianus were studied. Two doses of each probiotic (5 and 10 g kg^?1 of food) were added to the diets and fed to frogs, totalling five treatments over 112 days. Haematological analyses consisted of total and differential leucocyte counts, erythrocyte and thrombocyte counts, haematocrit, haemoglobin levels and RBC indices (mean corpuscular volume, mean corpuscular haemoglobin – and mean corpuscular haemoglobin concentration) and the immunological parameters included phagocytic capacity and phagocytic index of peritoneal phagocytes. The results showed that the probiotics did not significantly influence any of the haematological parameters measured. However, immunological assays showed that the probiotics had an immunostimulating effect. The greatest effects were seen with probiotic P₁ fed at a dose of 10 g kg^?1 of diet and probiotic P₂ fed at 5 g kg^?1 of diet.     

Effect of ascorbic acid supplementation on zootechnical performance,haematological parameters and sperm quality of lebranche mullet Mugil liza

The feed provided to breeding fish is one of the main factors influencing the quality of fish gametes. This study was carried out to evaluate the influence of ascorbic acid on growth, haematological parameters and sperm quality of Lebranche mullet males (Mugil liza). Six diets with different levels of ascorbic acid (0; 53; 107; 216; 482 and 708 mg/kg) were tested in triplicate for 75 days. During spermiation (first gonadal maturation), 144 individuals (205.7 ± 11.5 g and 25.7 ± 0.4 cm) were randomly distributed in 18 experimental tanks. Growth parameters were evaluated at the beginning and end of the experiment. Fish blood was collected to analyse glucose, total protein and erythrocyte count (EC) (n = 9). Fish (n = 12) from each treatment were euthanized to determine hepatosomatic index (HSI) and gonadosomatic index (GSI). Semen was collected to evaluate spermatic density, cell membrane integrity and sperm motility. No difference (p > .05) was found on growth parameters, GSI, HSI and total protein. However, EC was lower in fish fed without ascorbic acid (the control group). Ascorbic acid supplementation provided positive effects on sperm characteristics. Fish from treatments with 53 and 107 mg/kg presented the best results for motility time (133.30 ± 4.25 and 135.00 ± 2.77 respectively). Treatments with 107, 216 and 708 mg/kg provided the best results for motility rate (92.0 ± 2.9%, 93.0 ± 5.8% and 93.0 ± 5.8% respectively). Supplementation with 107 and 216 mg/kg provided the best results for plasma membrane integrity (70.12 ± 9.10% and 76.3 ± 3.1% respectively). Lower spermatic density was observed in fish without ascorbic acid supplementation, although no difference was found in sperm density among the treatments with ascorbic acid (p < .05). Considering these results, supplementation of dietary ascorbic acid between 107 and 216 mg/kg optimizes the spermatic quality in males of lebranche mullet.     

Improving the estimation of soil water evaporation based on days after wetting

Soil evaporation constitutes a major pathway of water loss in agriculture. Understanding its dynamics in the face of drying and soil cover is fundamental to improve both simulation models and the sustainability of production systems. Thus, the objective of this study was to estimate soil evaporation as a function of drying and percentage of soil cover. Three experiments were carried out in three different periods. In each of the periods, in parallel and in an adjacent area, an experiment was carried out to evaluate the influence of weighing micro-lysimeter (ML) height on the estimation of direct soil evaporation. The experiments were installed with a completely randomized design. For the experiments that measured evaporation as a function of cover, the treatments consisted of six different percentages of cover of the internal area of the ML (0%, 10%, 25%, 50%, 75% and 100%), with four replicates, using artificial plants to cover the soil. The experiments that estimated evaporation as a function of ML height were conducted with eight repetitions and three different ML heights (100, 200 and 300 mm). It was observed that the accumulated evaporation up to 25 days after wetting was 52% and 53% lower in micro-lysimeters with 100 mm height, compared to those with 200 and 300 mm height, respectively, for all experiments. The new model developed to calculate soil evaporation as a function of the days after wetting and percentage of soil cover showed excellent performance (NSE > 0.95).     

Are tree breeders properly predicting genetic gain? A case study involving <Emphasis Type="Italic">Corymbia</Emphasis> species

The estimation of quantitative genetic parameters in breeding programs is important to ensure efficient selection. In this context, knowledge of the mating system is critical, as it underpins assumptions about inter-relatedness on which variance component estimation depends. However, proper account of the breeding system is not always taken, either because it is unknown and/or because it is ignored. That eucalypts have a mixed-mating system is well-established, however many breeders use models that assume outcrossed mating with an infinite number of male parents (i.e. allogamous mating), from which genetic parameter estimates are then used to predict genetic gains. First-generation, open-pollinated progeny tests of Corymbia citriodora subsp. citriodora and C. citriodora subsp. variegata, being managed for seed production, were used to investigate the likely bias, resulting in overestimation of genetic parameters under the assumption of allogamous mating. When we assumed allogamous mating, we observed inflated predictions of additive variance and narrow-sense heritability (\(\hat{h}_{a}^{2}\)) of diameter at breast height and height. The overestimate of \(\hat{h}_{a}^{2}\) was approximately 32% for C. citriodora subsp. citriodora and 21% for C. citriodora subsp. variegata. Inappropriate modelling of relatedness in eucalypts that assumes panmictic outcrossing when in fact these species have a mixed-mating system results in overestimates of the population genetic gain with selection.     

Submicroscopy of Aggregates of Luvic Phaeozems under Different Land Uses in the Southeast of the Buenos Aires Province,Argentina

Eurasian Soil Science - Soil aggregates are key elements of soil structure that play an important role in several soil processes. One of the major factors that affect soil structure are the...     

Collection Rates and Morphology of Equine Oocytes Obtained from Immature Follicles after Treatment with Equine Pituitary Extract (EPE) and Human Chorionic Gonadotropin

This study examined the effect of treating mares with equine pituitary extract (EPE) in combination with human chorionic gonadotropin (hCG; EPE/hCG) on the recovery rate of immature oocytes by ovum pick-up (OPU) and on oocyte morphology. Ten mares were subjected to each of two treatments in a random sequence: superstimulated with EPE (25 mg, twice daily) and treated with hCG (2,500 IU) or control (no exogenous treatment). The cytoplasmic morphology of oocytes recovered was evaluated through transmission electron microscopy. Follicular fluid was collected at aspiration for progesterone analysis, which was performed by radioimmunoassay. The EPE/hCG did not increase the oocyte recovery rate from immature follicles when compared with the controls (15.5% and 16.7%, respectively). A significantly higher oocyte recovery rate per mare was observed (70% versus 50%). However, precocious granulosa cell expansion was observed with EPE/hCG treatment in contrast to the control (64.4% and 33% of follicles with expanded cumulus, respectively), and increased intrafollicular progesterone concentration was also seen (158.80 ng/mL versus 82.05 ng/mL). The ultrastructural analysis of oocytes from both groups showed morphologic features related to immaturity. Numerous vesicles containing cortical granules were found, distributed in clusters into the cytoplasm, and junctional complexes were still seen between oocyte and granulosa cells. In conclusion, EPE/hCG treatment induced some follicular modifications, but the recovery rate was not increased. All oocytes examined presented signs of immaturity.     

Pancytopenia in a cat with visceral leishmaniasis

Abstract: A 4‐year‐old, domestic shorthair, female spayed cat was presented for decreased appetite and depression. Severe pancytopenia with erythrocyte autoagglutination was found. The cat was seronegative for feline immunodeficiency and leukemia viruses. Immune‐mediated hemolytic anemia was suspected but no response to treatment with a blood transfusion, enrofloxacin, and prednisone was observed. Blood and bone marrow smears obtained 11 days later contained Leishmania amastigotes in the cytoplasm of neutrophils and macrophages, respectively. Serologic and PCR testing of peripheral blood confirmed infection with Leishmania infantum. Despite treatment, the cat worsened clinically and was euthanized. At necropsy, visceral dissemination of the parasite was confirmed. The findings in this case indicate that visceral leishmaniasis should be considered as a differential diagnoses in cats with pancytopenia in areas endemic for Leishmania. In addition, amastigotes may be observed in peripheral blood neutrophils.     

Dose-related antinociceptive effects of intravenous buprenorphine in cats

The dose-related antinociceptive effects of intravenous (IV) buprenorphine were evaluated in cats. Thermal (TT) and mechanical threshold (MT) devices were used for nociceptive stimulation. After baseline threshold recordings, buprenorphine was administered IV (0.01, 0.02 or 0.04 mg/kg; B1, B2 and B4, respectively) in a randomised, blinded and cross-over study. Data were analysed by ANOVA (P < 0.05) using 95% confidence intervals (CI). TT increased 15, 30, 45 min and 1 (5.2 ± 2.7 °C), 2, 3 and 4 h after B1; 15, 30, 45 min and 1 (5.1 ± 3.9 °C) and 2 h after B2, and 15, 30, 45 min and 1 (5.4 ± 3.3 °C), 2, 3, 6 and 8 h after B4. MT increased 15 and 45 min after B2 (260 ± 171 mmHg), and 30 (209 ± 116 mmHg) and 45 min and 1 and 2 h after B4. At 45 min, MT values were significantly higher after B2 compared to B1 (P < 0.05). With MT, B2 and B4 produced more antinociception and longer duration of action than B1, respectively. No dose response to thermal stimulation was detected.     

Silica content in soil solution and its relation with phytolith weathering and silica biogeochemical cycle in Typical Argiudolls of the Pampean Plain,Argentina—a preliminary study

Purpose  

Little is known on the silica biogeochemical cycle in terrestrial environments. The aim of this work is to assess phytolith’s role on the biogeochemical cycle of Si in Typical Argiudolls under different vegetation of the Pampean Plain, Argentina.