Effects of re-stripping on the seminal characteristics of pacu (Piaractus mesopotamicus) during the breeding season

Seminal characteristics in teleost fish with an annual reproductive period, such as pacu (Piaractus mesopotamicus), may vary during the breeding season. The sperm formed before the beginning of the spawning period may be stored for a long time, causing damage to the cells. Therefore, re-stripping may be an important way to eliminate the old and allow for the collection of new spermatozoids. In this study, we analyzed the seminal characteristics of hormonally induced pacu at the beginning, middle and end of the breeding season, and we analyzed samples from re-stripped males (stripped first at the beginning, re-stripped in the middle, and re-stripped again at the end of the season) during two breeding seasons. The sperm density, ionic composition, pH, and osmolality were similar among the groups. The semen volume, seminal plasma protein concentration and incidence of morphologically anomalous sperm increased over time. In addition, some parameters that are associated with good-quality semen decreased, such as sperm motility, viability and DNA integrity. Moreover, we observed a positive association among motility, viability and DNA integrity for sperm with elevated 11-ketotestosterone, but there was no such association for fshb or lhb mRNA levels in the pituitary. The semen that was obtained earlier (at the beginning) or from re-stripped males exhibited better characteristics than the other samples collected. In conclusion, collecting semen from pacu at the end of breeding season should be avoided; it is preferable to strip early and then re-strip later in the season, and this approach may be used for diverse aquaculture purposes.

Carga de trabalho de equinos da raça Quarto de Milha monitorados com Sistema de Posicionamento Global (GPS) e monitor cardíaco durante exercício de Três Tambores

Pós-graduação em Biotecnologia Animal - FMVZ

Supplementation with protected fat to manage gastro-intestinal nematode infection in Santa Ines sheep

This experiment aimed to determine the influence of supplementation with protected fat on nematode infections in periparturient sheep or sheep in their final stage of pregnancy. Fifty Santa Ines ewes received 200 g of concentrate/animal/day, mineral salt and water and were grouped according to their weight, hematocrit and the number of eggs per gram of feces (EPG). Two diets were used: a control treatment and a treatment consisting of supplementation with 30 g/animal/day of protected fat in the concentrate. Three monthly evaluations were performed over a total of 84 days of testing. The variables analyzed were weight, body condition, EPG and coprocultures; blood tests were performed for the determination of packed cell volume, hemoglobin concentration and total plasma protein, and leukograms and eosinophil counts were performed. For weight and hemoglobin concentrations, interactions were observed between diet and collection date (P<0.05); body condition, total plasma protein, packed cell volume and total leukocytes did not differ statistically among treatments (P> 0. 05) but did differ by collection day (P <0.05). The EPG and eosinophil counts did not differ statistically by either diet or collection date (P>0.05). The genus Haemonchus was predominant, followed by the genera Cooperia, Trichostrongylus and Oesophagostomum. The protected fat did not decrease EPG and did not improve the blood parameters of infected sheep.

Vitamin A affects haematology, growth and immune response of Nile tilapia (Oreochromis niloticus, L.), but has no protective effect against bacterial challenge or cold-induced stress

Vitamin A (vitA) is an essential nutrient that acts as an endocrine regulator of several metabolic pathways, modulating normal growth and health status of animals. Although the importance of vitA for normal haematology and immune response is well documented for higher vertebrates, there is limited information on the physiological effects of vitA on fish. Therefore, we designed a 130-day feeding trial to evaluate the effect of vitA supplementation on growth, haematology, immune function and resistance to experimental infection with Aeromonas hydrophila and cold-induced stress. A group of 320 Nile tilapia fingerlings 7.49 ± 0.19 g weight (mean ± SD) were randomly stocked into 40 250 L-aquaria and fed practical diets containing graded levels of vitA (0, 0.06, 0.12, 0.24, 0.48, 0.96, 1.92, 3.84 mg retinol (ROH) kg−1 diet. Growth, haematology, plasma protein profile and immune response were significantly affected by vitA supplementation; however, no clear protective effect of vitA supplementation on disease and cold stress resistance were observed in this study. Clinical signs of vitA deficiency were: resting and abnormal swimming behaviour, exophthalmia, haemorrhages at the base of fins and on skin, serous fluids in abdominal cavity, neutropenia, reduction in red blood cell count, haematocrit and haemoglobin evolving to high mortality rates in a short period of time. A dietary level of vitA around 1.2 mg ROH kg−1 may be required to prevent gross deficiency signs and promote proper growth and health status of Nile tilapia. VitA does not seem to have a pronounced effect on leucocyte differentiation, but clearly plays an important role on maintaining normal erythropoiesis.

Influência do tratamento anti-helmíntico e da suplementação no desempenho de bezerros nelore no período pós-desmame

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Acid-base and biochemical stabilization and quality of recovery in male cats with urethral obstruction and anesthetized with propofol or a combination of ketamine and diazepam

This study compared acid-base and biochemical changes and quality of recovery in male cats with experimentally induced urethral obstruction and anesthetized with either propofol or a combination of ketamine and diazepam for urethral catheterization. Ten male cats with urethral obstruction were enrolled for urethral catheterization and anesthetized with either ketamine-diazepam (KD) or propofol (P). Lactated Ringer's solution was administered by intravenous (IV) beginning 15 min before and continuing for 48 h after relief of urethral obstruction. Quality of recovery and time to standing were evaluated. The urethral catheter was maintained to measure urinary output. Hematocrit (Hct), total plasma protein (TPP), albumin, total protein (TP), blood urea nitrogen (BUN), creatinine, pH, bicarbonate (HCO3-), chloride, base excess, anion gap, sodium, potassium, and partial pressure of carbon dioxide in mixed venous blood (pvCO(2)) were measured before urethral obstruction, at start of fluid therapy (0 h), and at subsequent intervals. The quality of recovery and time to standing were respectively 4 and 75 min in the KD group and 5 and 16 min in the P group. The blood urea nitrogen values were increased at 0, 2, and 8 h in both groups. Serum creatinine increased at 0 and 2 h in cats administered KD and at 0, 2, and 8 h in cats receiving P, although the values were above the reference range in both groups until 8 h. Acidosis occurred for up to 2 h in both groups. Acid-base and biochemical stabilization were similar in cats anesthetized with propofol or with ketamine-diazepam. Cats that received propofol recovered much faster, but the ketamine-diazepam combination was shown to be more advantageous when treating uncooperative cats as it can be administered by intramuscular (IM) injection.

Is zinc-alpha 2-glycoprotein a cardiovascular protective factor for patients undergoing hemodialysis?

Background: Zinc-alpha 2-glycoprotein (ZAG) is a lipid mobilizing factor. Its anti-inflammatory action and expression pattern suggest that ZAG could act by protecting against the obesity-associated disorders. In hemodialysis (HD) patients, ZAG levels were described to be elevated but its effects on markers of inflammation and LDL oxidation are still unclear. We investigated the relationship between ZAG and markers of systemic inflammation and LDL atherogenic modification profile in HD patients. Methods: Forty-three patients regularly on HD were studied and compared to 20 healthy subjects. Plasma ZAG, adiponectin, electronegative LDL [LDL(-)], an atherosclerotic negatively charged LDL subtraction, and anti-LDL(-) autoantibodies levels were measured by ELISA. Markers of inflammation and atherogenic cell recruitment (TNF-alpha, interleukin-6, VCAM-1, ICAM-1, MCP-1 and PAI-1) were also determined. Results: Inflammatory markers and atherogenic cell recruitment were higher in HD patients when compared to healthy subjects. ZAG levels were also higher in HD patients (151.5 +/- 50.1 mg/l vs 54.6 +/- 23.0 mg/l; p<0.0001) and its levels were negatively correlated with TNF-alpha (r= -0.39; p = 0.001) and VCAM-1 (r= -0.52; p<0.0001) and, positively correlated with anti-LDL(-) autoantibodies (r = 038; p = 0.016). On multivariate analyses, plasma ZAG levels were independently associated with VCAM-1 (p = 0.01). Conclusion: ZAG is inversely associated with markers of pro-atherogenic factors linked to systemic inflammation and oxidative stress. Thus, this adipokine may constitute a novel marker of a favorable metabolic profile regarding cardiovascular risk factors in HD population. (C) 2011 Elsevier B.V. All rights reserved.

Hematologic Reference Values for Clinically Healthy Captive Golden Conures (Guaruba guarouba)

Golden conures or ararajubas (Guaruba guarouba) are endangered parrots endemic to the Brazilian Amazon forest. Body mass, blood cell counts, and total plasma protein were determined for 70 clinically healthy golden conures captive at zoologic parks and private breeder facilities in Brazil. Hematologic results (mean +/- SD) were: Erythrocytes 3.6 +/- 0.5 x 10(6) cells/mm(3), hemoglobin 12.8 +/- 1.4 g/dl, packed cell volume 46 +/- 3.8%, mean corpuscular volume 132 +/- 20 fl, mean corpuscular hemoglobin (MCH) 36 +/- 5.7 pg, mean corpuscular hemoglobin concentration (MCHC) 28 +/- 3.5%, thrombocytes 26.3 +/- 9.3 x 10(3) cells/mm(3), leukocytes 11.9 +/- 4.5 x 10(3) cells/mm(3), heterophils 6284 +/- 2715 cells/mm(3), lymphocytes 5473 +/- 2408 cells/mm(3), monocytes 113 +/- 162 cells/mm(3), eosinophils 10 +/- 42 cells/mm(3), basophils 27 +/- 64 cells/mm(3). Body mass was 254 +/- 24.9 g and total plasma protein (TPP) was 3.54 +/- 0.58 g/dl. No statistical differences were observed between genders within age groups. Differences between juveniles (J) and adults (A) were identified for TPP < A), MCH (J > A), and MCHC (J > A). These results provide reliable reference values for the clinical interpretation of hematologic results for the species. Hematology may be an important tool for population health investigations on free-ranging golden conure populations and will also be essential to survey the health of release candidates in future reintroduction programs.

Structure of the haptoglobin-haemoglobin complex

Red cell haemoglobin is the fundamental oxygen-transporting molecule in blood, but also a potentially tissue-damaging compound owing to its highly reactive haem groups. During intravascular haemolysis, such as in malaria and haemoglobinopathies(1), haemoglobin is released into the plasma, where it is captured by the protective acute-phase protein haptoglobin. This leads to formation of the haptoglobin-haemoglobin complex, which represents a virtually irreversible non-covalent protein-protein interaction(2). Here we present the crystal structure of the dimeric porcine haptoglobin-haemoglobin complex determined at 2.9 angstrom resolution. This structure reveals that haptoglobin molecules dimerize through an unexpected beta-strand swap between two complement control protein (CCP) domains, defining a new fusion CCP domain structure. The haptoglobin serine protease domain forms extensive interactions with both the alpha- and beta-subunits of haemoglobin, explaining the tight binding between haptoglobin and haemoglobin. The haemoglobin-interacting region in the alpha beta dimer is highly overlapping with the interface between the two alpha beta dimers that constitute the native haemoglobin tetramer. Several haemoglobin residues prone to oxidative modification after exposure to haem-induced reactive oxygen species are buried in the haptoglobin-haemoglobin interface, thus showing a direct protective role of haptoglobin. The haptoglobin loop previously shown to be essential for binding of haptoglobin-haemoglobin to the macrophage scavenger receptor CD163 (ref. 3) protrudes from the surface of the distal end of the complex, adjacent to the associated haemoglobin alpha-subunit. Small-angle X-ray scattering measurements of human haptoglobin-haemoglobin bound to the ligand-binding fragment of CD163 confirm receptor binding in this area, and show that the rigid dimeric complex can bind two receptors. Such receptor cross-linkage may facilitate scavenging and explain the increased functional affinity of multimeric haptoglobin-haemoglobin for CD163 (ref. 4).

Enhancement of blood-tumor barrier permeability by Sar-[D-Phe8]des-Arg9BK, a metabolically resistant bradykinin B1 agonist, in a rat C6 glioma model

Abstract Background While it is well known that bradykinin B2 agonists increase plasma protein extravasation (PPE) in brain tumors, the bradykinin B1 agonists tested thus far are unable to produce this effect. Here we examine the effect of the selective B1 agonist bradykinin (BK) Sar-[D-Phe8]des-Arg9BK (SAR), a compound resistant to enzymatic degradation with prolonged activity on PPE in the blood circulation in the C6 rat glioma model. Results SAR administration significantly enhanced PPE in C6 rat brain glioma compared to saline or BK (p < 0.01). Pre-administration of the bradykinin B1 antagonist [Leu8]-des-Arg (100 nmol/Kg) blocked the SAR-induced PPE in the tumor area. Conclusions Our data suggest that the B1 receptor modulates PPE in the blood tumor barrier of C6 glioma. A possible role for the use of SAR in the chemotherapy of gliomas deserves further study.

Plasmonic nanoparticles as sensors : a novel approach to quantify adsorption and diffusion kinetics

Biosensors find wide application in clinical diagnostics, bioprocess control and environmental monitoring. They should not only show high specificity and reproducibility but also a high sensitivity and stability of the signal. Therefore, I introduce a novel sensor technology based on plasmonic nanoparticles which overcomes both of these limitations. Plasmonic nanoparticles exhibit strong absorption and scattering in the visible and near-infrared spectral range. The plasmon resonance, the collective coherent oscillation mode of the conduction band electrons against the positively charged ionic lattice, is sensitive to the local environment of the particle. I monitor these changes in the resonance wavelength by a new dark-field spectroscopy technique. Due to a strong light source and a highly sensitive detector a temporal resolution in the microsecond regime is possible in combination with a high spectral stability. This opens a window to investigate dynamics on the molecular level and to gain knowledge about fundamental biological processes.rnFirst, I investigate adsorption at the non-equilibrium as well as at the equilibrium state. I show the temporal evolution of single adsorption events of fibrinogen on the surface of the sensor on a millisecond timescale. Fibrinogen is a blood plasma protein with a unique shape that plays a central role in blood coagulation and is always involved in cell-biomaterial interactions. Further, I monitor equilibrium coverage fluctuations of sodium dodecyl sulfate and demonstrate a new approach to quantify the characteristic rate constants which is independent of mass transfer interference and long term drifts of the measured signal. This method has been investigated theoretically by Monte-Carlo simulations but so far there has been no sensor technology with a sufficient signal-to-noise ratio.rnSecond, I apply plasmonic nanoparticles as sensors for the determination of diffusion coefficients. Thereby, the sensing volume of a single, immobilized nanorod is used as detection volume. When a diffusing particle enters the detection volume a shift in the resonance wavelength is introduced. As no labeling of the analyte is necessary the hydrodynamic radius and thus the diffusion properties are not altered and can be studied in their natural form. In comparison to the conventional Fluorescence Correlation Spectroscopy technique a volume reduction by a factor of 5000-10000 is reached.

The adsorption of biomolecules to multi-walled carbon nanotubes is influenced by both pulmonary surfactant lipids and surface chemistry

Background During production and processing of multi-walled carbon nanotubes (MWCNTs), they may be inhaled and may enter the pulmonary circulation. It is essential that interactions with involved body fluids like the pulmonary surfactant, the blood and others are investigated, particularly as these interactions could lead to coating of the tubes and may affect their chemical and physical characteristics. The aim of this study was to characterize the possible coatings of different functionalized MWCNTs in a cell free environment. Results To simulate the first contact in the lung, the tubes were coated with pulmonary surfactant and subsequently bound lipids were characterized. The further coating in the blood circulation was simulated by incubating the tubes in blood plasma. MWCNTs were amino (NH2)- and carboxyl (-COOH)-modified, in order to investigate the influence on the bound lipid and protein patterns. It was shown that surfactant lipids bind unspecifically to different functionalized MWCNTs, in contrast to the blood plasma proteins which showed characteristic binding patterns. Patterns of bound surfactant lipids were altered after a subsequent incubation in blood plasma. In addition, it was found that bound plasma protein patterns were altered when MWCNTs were previously coated with pulmonary surfactant. Conclusions A pulmonary surfactant coating and the functionalization of MWCNTs have both the potential to alter the MWCNTs blood plasma protein coating and to determine their properties and behaviour in biological systems.

Biocompatibility assessment of the first generation PediaFlow pediatric ventricular assist device

The PediaFlow pediatric ventricular assist device is a miniature magnetically levitated mixed flow pump under development for circulatory support of newborns and infants (3-15 kg) with a targeted flow range of 0.3-1.5 L/min. The first generation design of the PediaFlow (PF1) was manufactured with a weight of approximately 100 g, priming volume less than 2 mL, length of 51 mm, outer diameter of 28 mm, and with 5-mm blood ports. PF1 was evaluated in an in vitro flow loop for 6 h and implanted in ovines for three chronic experiments of 6, 17, and 10 days. In the in vitro test, normalized index of hemolysis was 0.0087 ± 0.0024 g/100L. Hemodynamic performance and blood biocompatibility of PF1 were characterized in vivo by measurements of plasma free hemoglobin, plasma fibrinogen, total plasma protein, and with novel flow cytometric assays to quantify circulating activated ovine platelets. The mean plasma free hemoglobin values for the three chronic studies were 4.6 ± 2.7, 13.3 ± 7.9, and 8.8 ± 3.3 mg/dL, respectively. Platelet activation was low for portions of several studies but consistently rose along with observed animal and pump complications. The PF1 prototype generated promising results in terms of low hemolysis and platelet activation in the absence of complications. Hemodynamic results validated the magnetic bearing design and provided the platform for design iterations to meet the objective of providing circulatory support for young children with exceptional biocompatibility.

PAPP-A and osteoprotegerin, together with interleukin-8 and RANTES, are elevated in the peritoneal fluid of women with endometriosis

OBJECTIVE: The purpose of this study was to evaluate whether pregnancy-associated plasma protein A, glycodelin, osteoprotegerin, and soluble CD163 are possible peritoneal fluid markers for endometriosis and to compare them with the established chemokine markers interleukin-8 and regulated on activation, normal T-cell expressed and secreted. STUDY DESIGN: Determination of the concentrations of interleukin-8, regulated on activation, normal T-cell expressed and secreted, pregnancy-associated plasma protein A, glycodelin, CD163, osteoprotegerin, and progesterone in the peritoneal fluid collected from women undergoing laparoscopy. RESULTS: From a total of 132 women, 77 women were diagnosed with endometriosis, and 55 women were free of the disease and served as control subjects. Pregnancy-associated plasma protein A and osteoprotegerin showed significantly (P < 0.05) elevated peritoneal fluid concentrations as a function of the severity of the disease, together with interleukin-8 and regulated on activation, normal T-cell expressed and secreted (P < .001). Glycodelin and CD163 did not differ between cases and control subjects. Many of these marker concentrations were intercorrelated strongly. CONCLUSION: Pregnancy-associated plasma protein A and osteoprotegerin may play a role in the inflammation process of endometriosis, but interleukin-8 and regulated on activation, normal T-cell expressed and secreted are superior peritoneal fluid markers.

Inflammation and NO(X)-induced nitration: assay for 3-nitrotyrosine by HPLC with electrochemical detection

The identification of 15N-labeled 3-nitrotyrosine (NTyr) by gas chromatography/mass spectroscopy in protein hydrolyzates from activated RAW 264.7 macrophages incubated with 15N-L-arginine confirms that nitric oxide synthase (NOS) is involved in the nitration of protein-bound tyrosine (Tyr). An assay is presented for NTyr that employs HPLC with tandem electrochemical and UV detection. The assay involves enzymatic hydrolysis of protein, acetylation, solvent extraction, O-deacetylation, and dithionite reduction to produce an analyte containing N-acetyl-3-aminotyrosine, an electrochemically active derivative of NTyr. We estimate the level of protein-bound NTyr in normal rat plasma to be approximately 0-1 residues per 10(6) Tyr with a detection limit of 0.5 per 10(7) Tyr when > 100 nmol of Tyr is analyzed and when precautions are taken to limit nitration artifacts. Zymosan-treated RAW 264.7 cells were shown to have an approximately 6-fold higher level of protein-bound NTyr compared with control cells and cells treated with N(G)-monomethyl-L-arginine, an inhibitor of NOS. Intraperitoneal injection of F344 rats with zymosan led to a marked elevation in protein-bound NTyr to approximately 13 residues per 10(6) Tyr, an approximately 40-fold elevation compared with plasma protein of untreated rats; cotreatment with N(G)-monomethyl-L-arginine inhibited the formation of NTyr in plasma protein from blood and peritoneal exudate by 69% and 53%, respectively. This assay offers a highly sensitive and quantitative approach for investigating the role of reactive byproducts of nitric oxide in the many pathological conditions and disease states associated with NO(X) exposure such as inflammation and smoking.