Características hematológicas, bioquímicas e biométricas de Piaractus mesopotamicus Holmberg, 1887 (Osteichthyes: Characidae) oriundos de cultivo intensivo

Hematological characteristics are important tools for diagnosis and prognostic features in fish population and they contribute to the comprehension of comparative physiology, phylogenetic relation, food habit and other ecological parameters. Red blood cell count (RBC), white blood cell count (WBC), thrombocytes counts, glucose, cortisol, hepatosomatic and splenosomatic relation were evaluated in Piaractus mesopotamicus. The values of hepatosomatic relation varied from 0.67% to 2.79% and splenosomatic relation from 0.011% to 0.108%. RBC 1.870 to 4.590 × 106/μL, hemoglobin concentration 5.7 to 12.1g/dL, haematocrit 24.0% to 40.0%, mean corpuscular volume 98.0 to 174.0 fL, mean corpuscular haemoglobin concentration 19.1 to 34.2g/dL, glucose 40.6 to 89.2mg/dL and cortisol 4.73 to 110.5ng/dL The number of total thrombocytes was 12,160.0 to 89,280.0/μL and WBC 2,019.0 to 47,473.0/μL. Differential count of leucocytes showed 54.1% limphocytes, 10.9% neutrophils, 10.3% monocytes, 9.9% PAS-positive granular leukocytes (PAS-GL), 6.7% eosinophils, 0.3% basophils and 7.8% immature leucocytes. Variation of these haematological characteristics in the same fish species was observed, being considered as the normal variation in the fish concerned. Hence it is incorrect to assume that biological characteristics in fishes are distributed normally.

Critical protective role for annexin 1 gene expression in the endotoxemic murine microcirculation

The inflammatory response is a protective process of the body to counteract xenobiotic penetration and injury, although in disease this response can become deregulated. There are endogenous biochemical pathways that operate in the host to keep inflammation under control. Here we demonstrate that the counter-regulator annexin 1 (AnxA1) is critical for controlling experimental endotoxemia. Lipopolysaccharide (LPS) markedly activated the AnxA1 gene in epithelial cells, neutrophils, and peritoneal, mesenteric, and alveolar macrophages-cell types known to function in experimental endotoxemia. Administration of LPS to AnxA1-deficient mice produced a toxic response characterized by organ injury and lethality within 48 hours, a phenotype rescued by exogenous application of low doses of the protein. In the absence of AnxA1, LPS generated a deregulated cellular and cytokine response with a marked degree of leukocyte adhesion in the microcirculation. Analysis of LPS receptor expression in AnxA1-null macrophages indicated an aberrant expression of Toll-like receptor 4. In conclusion, this study has detailed cellular and biochemical alterations associated with AnxA1 gene deletion and highlighted the impact of this protective circuit for the correct functioning of the homeostatic response to sublethal doses of LPS. Copyright © American Society for Investigative Pathology.

Spatial and temporal profiles for anti-inflammatory gene expression in leukocytes during a resolving model of peritonitis

The recent appreciation of the role played by endogenous counterregulatory mechanisms in controlling the outcome of the host inflammatory response requires specific analysis of their spatial and temporal profiles. In this study, we have focused on the glucocorticoid-regulated anti-inflammatory mediator annexin 1. Induction of peritonitis in wild-type mice rapidly (4 h) produced the expected signs of inflammation, including marked activation of resident cells (e.g., mast cells), migration of blood-borne leukocytes, mirrored by blood neutrophilia. These changes subsided after 48-96 h. In annexin 1null mice, the peritonitis response was exaggerated (∼40% at 4 h), with increased granulocyte migration and cytokine production. In blood leukocytes, annexin 1 gene expression was activated at 4, but not 24, h postzymosan, whereas protein levels were increased ai both time points. Locally, endothelial and mast cell annexin 1 gene expression was not detectable in basal conditions, whereas it was switched on during the inflammatory response. The significance of annexin 1 system plasticity in the anti-inflammatory properties of dexamethasone was assessed. Clear induction of annexin 1 gene in response to dexamethasone treatment was evident in the circulating and migrated leukocytes, and in connective tissue mast cells; this was associated with the steroid failure to inhibit leukocyte trafficking, cytokine synthesis, and mast cell degranulation in the annexin 1null mouse. In conclusion, understanding how inflammation is brought under control will help clarify the complex interplay between pro- and anti-inflammatory pathways operating during the host response to injury and infection. Copyright © 2006 by The American Association of Immunologists, Inc.

Morphological, cytochemical, and ultrastructural study of thrombocytes and leukocytes in neotropical fish, Brycon orbignyanus Valenciennes, 1850 (Characidae, Bryconinae)

Morphological, cytochemical and ultrastructural studies are important to demonstrate the function of the blood cells, which is very little understood in teleosts. In peripheral blood of piracanjuba' Brycon orbignyanus, thrombocytes, lymphocytes, monocytes, neutrophils and heterophils were studied and characterized. Thrombocytes had a fusiform or oval shape with PAS-positive granules. Lymphocytes presented small size with sparse basophilic cytoplasm. Monocytes were large in size, presented basophilic cytoplasm that may be foamy or vacuolated, with non-specific esterase staining. The neutrophils presented lightly neutrophilic granule cytoplasm, with positivity for PAS and peroxidase. The heterophils were large in size, with eosinophilic and basophilic granules cytoplasm and PAS-positive. Transmission electron microscopy study demonstrated that the thrombocytes, lymphocytes and monocytes features were similar to other teleosts. In ultrastructural study only one type of neutrophils was observed. Cytochemical findings indicated that neutrophils and monocytes of B. orbignyanus may be involved in phagocytosis, and neutrophils play an important microbicidal role.

The influence of dietary vitamin C and E supplementation on the physiological response of pirarucu, Arapaima gigas, in net culture

This study evaluated the efficacy of dietary vitamin C (ascorbic acid or AA), vitamin E (α-tocopherol or α-T), and C + E supplementation on the blood parameters of Arapaima gigas grown in net cages for 45 days. Four treatments were tested: control (commercial feed); C800; E500 and C + E (800 + 500) with supplementation of 800 mg AA kg- 1, 500 mg α-T kg- 1 and 800 + 500 mg AA + α-T kg- 1, respectively. Hematocrit (Ht), red blood cells (RBC), and hemoglobin concentration (Hb) (oxidative status indicators), thrombocytes and leukocytes (immunological indicators), plasma protein and glucose were evaluated. Fish fed vitamin C and C + E supplemented diets showed greater weight gain and survival. Dietary vitamin C and C + E diet supplementation resulted in increased Ht, Hb, RBC, MCHC, total leukocytes, total proteins, thrombocytes and eosinophils compared to the control and α-T. The α-tocopherol-supplemented diet reduced the number of total thrombocytes, lymphocytes and neutrophils and increased glucose and eosinophils relatively to the control. In general, leukocytes and thrombocytes were good indicators of the efficiency of vitamin on the defense mechanism of the A. gigas reared in cages. Results indicate that high α-T diet supplementation provides no benefit for the maintenance of the oxidative or the immunological status of A. gigas. However, it was demonstrated that high dietary AA improves A. gigas immunological status. Red blood cell indices and immune system indicators showed no synergistic effect between the vitamins after supplementing the A. gigas diet with α-T + AA. © 2006 Elsevier Inc. All rights reserved.

Tissue inhibitor of metalloproteinase-2 (TIMP-2) location in the ventral, lateral, dorsal and anterior lobes of rat prostate by immunohistochemistry

Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play a major role in extracellular matrix component degradation in several normal and abnormal tissue situations; they are also found in human seminal plasma. MMPs have been found in rat prostate secretions and are nearly lobe specific in expression pattern. The aim of this study was to evaluate whether TIMP-2, like other semen components, is expressed differently from different rat prostatic lobes. Immunohistochemical staining was performed in both young and adult rat ventral (VP), lateral (LP), dorsal (DP), and anterior (AP) prostatic lobes and confirmed by western blotting. TIMP-2 expression was found in the epithelial cells in the following sequence: LP > AP > DP > VP, in both young and adult rats. In this study, 100% of adult LP presented histological signs of prostatitis, where TIMP-2 immunostaining was positive in normal epithelium even with intraluminal neutrophils, but was reduced or absent in the epithelium with intraepithelial leukocytes or with periductal stroma disorganization associated with mononuclear cell infiltration. However, TIMP-2 expression in LP was not induced by prostatitis, since younger rat LPs were also strongly TIMP-2 positive. The distal and intermediate VP regions were TIMP-2 negative, but the proximal regions were strongly stained. Western blotting results confirmed the high TIMP-2 expression in the LP lobe. Thus, TIMP-2 is expressed differently between the prostatic lobes and is another nearly lobe-specific protein, which plays a role in the regulation of MMP activity in seminal plasma and glandular homeostasis. TIMP-2 is also another regional ductal variation of VP. Further studies should address whether TIMP-2 expression is related to the highest incidence of rat LP prostatitis and adenocarcinoma. © 2006 International Federation for Cell Biology.

Antisperm antibodies in infertile men and their correlation with seminal parameters

Aim: Antisperm antibodies (ASA) in males cause the autoimmune disease 'immune infertility'. The present study intended to detect the presence of ASA and their incidence in men with unexplained infertility, as well as to evaluate the correlation between the presence of ASA and semen parameter alterations. Methods: Blood and sperm assessment were collected to carry out a direct and indirect mixed antiglobulin reaction (MAR) test and semen analysis in infertile and fertile men from the University Hospital of the Faculty of Medicine, Sao Paulo State University, Sao Paulo. Results: In the MARtest, 18.18% of infertile men were positive for ASA. In fertile men, no positivity was found. A significant correlation between the presence of ASA with an increased white blood cell count plus a decreased hypoosmotic swelling test result was observed. Conclusions: The results indicate that ASA are involved in reduced fertility. It is not ASA detection per.se that provides conclusive information about the occurrence of damage to fertility. The correlation between infertility and altered seminal parameters reinforce the ASA participation in this pathology. © 2007 The Authors Journal compilation. © 2007 Japan Society for Reproductive Medicine.

Does cotinine act upon reactive oxygen species and peroxidases?

Nicotine, an oxidizing agent, is certainly one of the most widely used alkaloids in the world. It is, together with its main metabolite, cotinine, responsible for tobacco-dependence. The use of tobacco is closely associated with lung disease, morphological leukocyte modification and generation of oxidant species. The aim of this study was to look for a possible relationship between cotinine, oxidant species generation and oxidative processes. After studying the action of cotinine in some chemical oxidation models and on the enzymatic kinetics of peroxidases (myeloperoxidase and horseradish peroxidase), we concluded that cotinine does not act directly upon H 2O 2, HOCl, taurine chloramines, horseradish peroxidase or myeloperoxidase.

The effect of age on hematological and serum biochemical values on juvenile ring-necked pheasants (Phasianus colchicus)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Influence of sex, age, and fasting on blood parameters and body, bursa, spleen and yolk sac weights of broiler chicks

The effects of water and feed fasting for 24, 48 and 72 hours post-hatching on blood parameters (mean corpuscular volume, MCV; red blood-cell, RBC; hematocrit, HCT; hemoglobin, HGB; plasma glucose, CGP; plasma total protein, PP, and differential leukocytes count), and on body, liver, spleen, bursa, and yolk sac weights were analyzed. Erythrogram data were obtained with a blood cell counter. Total plasma protein and plasma glucose were determined by using the Bradford method (1976) and a glucose PAP liquiform kit (Labtest, cat. n. 84), respectively. Specific leukocyte counts were carried out on blood smears stained with Rosenfeld solution. According to the obtained data, water and feed post-hatching fasting reduced MCV values, which also were lower in males than that in females. Fasting for 48 hours promoted an increase in PP, while fasting for 72 hours reduced HCT. Chicks submitted to fasting presented lower body weights as compared to fed chicks, but their liver weight did not increase between 48 and 72 hours of age. Fasting decreased spleen weight, but bursa and yolk sac weight were not affected. Data showed that female and male chicks react in a similar way to post-hatching fasting, which affects body weight, liver and spleen weight, and HCT and PP values. Moreover, 72 hours of fasting affected more intensely HCT and MCV values.

Morphological characterization of the leukocytes in circulating blood of the turtle (Phrynops hilarii)

The Phrynops hilarii specie of turtle has its characterization not well defined in the literature, it was proposed in this study the leukocyte characterization of the blood, stained by Leishman and analyzed under light and transmission electron microscope. It was not observe any cellular type with similar characteristics to neutrophils in mammalian group. We believed, based on the data obtained in this study that the heterophils have a morphofuncional analogy with another neutrophils belonged to mammalian group. This conclusion is being supported in many recent studies found in the literature.

Human eosinophil adhesion and degranulation stimulated with eotaxin and RANTES in vitro: Lack of interaction with nitric oxide

Background: Airway eosinophilia is considered a central event in the pathogenesis of asthma. The toxic components of eosinophils are thought to be important in inducing bronchial mucosal injury and dysfunction. Previous studies have suggested an interaction between nitric oxide (NO) and chemokines in modulating eosinophil functions, but this is still conflicting. In the present study, we have carried out functional assays (adhesion and degranulation) and flow cytometry analysis of adhesion molecules (VLA-4 and Mac-1 expression) to evaluate the interactions between NO and CC-chemokines (eotaxin and RANTES) in human eosinophils. Methods: Eosinophils were purified using a percoll gradient followed byimmunomagnetic cell separator. Cell adhesion and degranulation were evaluated by measuring eosinophil peroxidase (EPO) activity, whereas expression of Mac-1 and VLA-4 was detected using flow cytometry. Results: At 4 h incubation, both eotaxin (100 ng/ml) and RANTES (1000 ng/ml) increased by 133% and 131% eosinophil adhesion, respectively. L-NAME alone (but not D-NAME) also increased the eosinophil adhesion, but the co-incubation of L-NAME with eotaxin or RANTES did not further affect the increased adhesion seen with chemokines alone. In addition, L-NAME alone (but not D-NAME) caused a significant cell degranulation, but it did not affect the CC-chemokine-induced cell degranulation. Incubation of eosinophils with eotaxin or RANTES, in absence or presence of L-NAME, did not affect the expression of VLA-4 and Mac-1 on eosinophil surface. Eotaxin and RANTES (100 ng/ml each) also failed to elevate the cyclic GMP levels above baseline in human eosinophils. Conclusion: Eotaxin and RANTES increase the eosinophil adhesion to fibronectin-coated plates and promote cell degranulation by NO-independent mechanisms. The failure of CC-chemokines to affect VLA-4 and Mac-1 expression suggests that changes in integrin function (avidity or affinity) are rather involved in the enhanced adhesion. © 2008 Lintomen et al; licensee BioMed Central Ltd.

Hematology of the bronze Turkey (Meleagris gallopavo): Variations with age and gender

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Tratamento hepatoprotetor favorece a resposta leucocitária de ratos wistar intoxicados por CCL4

The aims of this investigation was to evaluate the effect of hepatoprotective treatments with a compound prepared by the association of N-Acetyl DL-Methionine (5%) + Choline chloride (2%) + Caffeine (1%) + Thiamine hydrochloride (1%) + Nicotinamida (0,5%)+ Pyridoxine hydrochloride (0.04%), administered through intramuscular (IM) route, at doses of 0.2, 0.6 and 1.0 mL/kg of BW, through the study of leukocytes responses in rats submitted to acute intoxication with CCl4. 147 females were randomized into 21 groups, performing five different treatments, which were evaluated seven animals in four periods: two, four, six and eight days after CCl4-induced intoxication. In this study, it was observed absolute eosinophilia and monocytosis in animals untreated and treated with the lowest dose of 0.2 mL. These responses were significantly better in animals treated with 0.6 and 1.0 mL/ kg BW. The untreated animals showed thrombocytopenia, when compared to treated animals. Absolute neutropenia and lymphocytosis was observed in all rats intoxicated with CCl4, there is no difference among treatments. The analysis of white blood cells demonstrated that the hepatoprotective treatments favored the leukocyte response, by act beneficially on the population of these cells, supporting the hypothesis that these events may reduce the deleterious effects in liver tissue after intoxication by CCl4.

Diferentes fontes proteicas em rações de leitões sobre atividade da tripsina e parâmetros sangüíneos

Were used 64 piglets submitted eight treatments: ration with skim milk (SM), three rations with crescent levels of swine plasma (SP), three rations with whole egg (WE) and a ration with high inclusion of soybean meal (SB). Were monitored the blood parameters (BP) in pigs at 27 and at 34 days of age. The piglets were slaugther at 28 and at 35 days of age, for collections pancreas and posterior mensurements of absolut (AW) and relative weigth (RW) of pancreas and trypsin activity (TA). Treatments not influencied AW and TA. Significant effect of the crescent levels was verified of SP, with lineal reduction of the leukocytes and increase of the globular volume, to the 27 days; while to the 34 days, lineal increase of the hematias was observed. At 27 days, animals feds rations with crescent levels of SP have inferior percentage of eosinophils than others that consumed crescent levels of WE. The utilization of SP promoted smaller stimulus to the immune reply, while the use of WE promoted larger humoral reply of the piglets.