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.     

Cellular activity and development of enteric,hepatic and muscle tissues of juvenile pacu Piaractus mesopotamicus (Holmberg 1887) fed with lyophilized bovine colostrum

Bovine colostrum, an alternative food of animal origin provided as partial protein source, may constitute an innovative ingredient in fish feeding. Serum insulin‐like growth factor 1 (IGF‐1) and cellular activity of enteric, hepatic and muscle tissues of juvenile pacu Piaractus mesopotamicus fed with lyophilized bovine colostrum (LBC) were evaluated. Pacu (8.46 ± 0.74 g; 7.75 ± 0.27 cm, n = 18 in triplicate) were stocked into 18 tanks (300 L; 18 fish per tank, in triplicate) and fed with control diet containing 0% of LBC or experimental diet containing 10% and 20% of LBC inclusion for either 30 or 60 days of experiment. IGF‐1 serum concentration was not affected (P > 0.05) by the diet containing LBC and experimental period. The intestinal and hepatic cellular activity was not influenced by LBC (P > 0.05); however, intestine protein/RNA decreased at 60 days (P < 0.05). The muscle cellular activity was affected by LBC, highest in fish fed with diet containing 10% of LBC (P < 0.05). Between the periods DNA and protein/RNA increased at 60 days, while RNA, protein/DNA and RNA/DNA ratio decreased in this same period (P < 0.05). Increased DNA muscle content indicates fish growth by hyperplasia. The results of the present study is related with suitable development of enteric, hepatic and muscle tissues and proper condition of juvenile pacu fed with LBC considering the absence of differences in relation to juveniles that consumed diet without colostrum.     

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.     

Application of morphometric measures in estimation of body weight and discrimination of Astyanax lacustris and Astyanax fasciatus

The objective of this study was to verify the direct and indirect correlation between morphometric measures, ratios, body weight and yield in two lambari species Astyanax lacustris and Astyanax fasciatus and whether the discriminant analysis is capable of separating and allocating the species. We used 102 lambari yellow tail and 60 lambari red tail. The fish were weighed and submitted to the evaluation of the morphometric measurements. The direct and indirect effects were evaluated by the method of track analysis, considering weight at slaughter, weight of body parts and body yields as dependent variables and measures and morphometric ratios as explanatory variables. Astyanax lacustris presented higher height and body width, carcass yield and trunk, while A. fasciatus presented higher head yield and viscera weight. The discriminant analysis was able to classify 79.5% of the two species. Track analysis demonstrated that the morphometric measurements can be used for estimation of body and body components’ weight in A. lacustris and A. fasciatus. However, regarding the corporal yield; the morphometric measures were insufficient to explain the yield variations of the species.     

Nile tilapia broodfish fed high‐protein diets: Digestive enzymes in eggs and larvae

The aim of this study was to assess the activity of gastric, pancreatic and intestinal digestive enzymes during the embryonic and larval development of Nile tilapia (Oreochromis niloticus) GIFT strain Aqua America^® 1 obtained from a broodstock fed two levels of crude protein (CP). A total of 72 females and 24 males, 10 hapas, two CP levels (32% and 38%) and six phases of embryonic (cleavage, blastula, gastrula) and larval (hatching, 7 and 10 days post hatch, dph) stages were used. The eggs were collected in cleavage, blastula and gastrula stages, 300 mg was collected, and kept in cryogenic tubes in liquid nitrogen. For the samples at larval stage, the remaining eggs were separated according to crude protein level and kept in hatcheries and samples were collected on 7 and 10 dph the same way as before. A total of 48 samples were collected: at each protein level (32% and 38% CP), four samples were collected in each phase of embryonic and larval development. Statistical differences were not observed during embryonic development for acid proteases, trypsin, amylase and lipase activity at both levels of crude protein (32 and 38% CP). Differences for acid proteases were noticed on 7 dph; trypsin and amylase on 7 dph and 10 dph. Significant differences on blastula and 7 dph for protease; as for aminopeptidase, there was significant difference on 7 dph. The data indicated early appearance of digestive enzymes in Nile tilapia broodfish receiving 32% CP taking into account the rapid growth and development of this species.     

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.     

Combining ability of common bean parents in different seasons,locations and generations

The objectives of this study were to obtain information about genetic parameters of a partial diallel in common bean, as well as to evaluate the effects of growing seasons, locations and generations in these estimates in order to define the best strategies for the use of diallel analysis as methodology of selection of parents and populations in common bean breeding. Twelve parents were crossed in a partial diallel scheme (5?×?7), and the F₁ and F₂ generations were evaluated in four experiments (I, II, III, and IV). The effects of seasons were determined in the Experiments I and II, in which the F₁ generation was evaluated in two distinct growing seasons. To quantify the effect of locations, we used Experiments III and IV, in which the F₂ generation was evaluated in the same season, but at different locations, and to quantify the effect of generations, the experiments II (generation F₁) and III (F₂ generation) were evaluated in the same season and location. As for grain yield the genetic control varied according to the environment. An effect of interaction on the genetic parameters of the diallel was observed for generation, whereas for grain yield, effects of season and location were observed. For plant architecture, parental combining ability should therefore be estimated using the F₁ generation, while for grain yield, both the F₁ and F₂ can be used. The recombination between the best hybrids is a promising strategy for extraction of superior black bean lines.     

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...