Characterization and quantification of blood cells from the umbilical cord of dogs

Background: Models for the study of hematopoietic stem cells in dogs provide important information for bone marrow transplantation in humans. Recent studies have reported the importance of human umbilical cord blood (UCB) as an alternative to allogenic bone marrow for hematopoietic reconstitution. However, there are no studies on the UCB cells of dogs. Objective: the aim of this experiment was to characterize and quantify the blood cells of the umbilical cord of dogs. Methods: the blood of the umbilical cord of 20 neonatal dogs, delivered at term, with a median gestation time of 58 days, was collected with a 5-mL syringe containing EDTA. Total RBC, WBC, and platelet counts, HCT, hemoglobin (Hgb) concentration, and RBC indices were determined using an automatic cell counter. The differential leukocyte count was determined manually in blood smears stained with May-Grunwald-Giemsa. Reticulocyte percentages were determined on blood smears stained with brilliant cresyl blue and counterstained with May-Grunwald Giemsa. Results: the MCHC and numbers of RBCs, WBCs, neutrophils, and eosinophils in UCB were lower as compared with reference values for the peripheral blood of healthy neonatal and adult dogs; whereas, the MCV and reticulocyte percentages were higher. Conclusion: Erythrocyte macrocytosis and hypochromasia in UCB were consistent with marked reticulocytosis and indicative of high erythropoietic activity. The results of this study are an important first step in the characterization of UCB from neonatal dogs.

Genetic Parameters For the Somatic Cells Count In the Milk of Buffaloes Using Ordinary Test Day Models

The buffaloes dairy milk production (BDMP) has increased in the last 20 years, mainly for the manufacturing of mozzarella cheese, which is recognized by its high nutritional quality. However, this quality can be affected by several factors i. e. high somatic cells count (SCC) provokes changes in the milk's constituents. As in bovine dairy milk, the SCC is used as diagnostic tool for milk quality; because it enables the diagnosis of sub-clinic mastitis and also allows the selection of individuals genetically resistant to that disease. Based on it, we collected information about SCC and BDMP along the lactation in Murrah breed buffaloes, during the period between 1997 and 2005. Curves were designed to estimate genetic parameters. These parameters were estimated by ordinary test-day models. There were observed variations in the estimated heritability for both characteristics the lowest score for somatic cells count (SSCC) was seen at first month (0.01) and the highest at sixth months (0.29 the genetic correlation between these traits varied from -1 at the 1 and 9(th) months to 0.31 and 0.30 in the2 and 4(th) month of lactation. Phenotypic correlations were all negative (-0.07 in the second month and up to -0.35 in the eighth month of lactation). These results showed that environmental factors are more important than genetics in explain SCC, for this reason, selection for genetic resistance to mastitis in buffalos based in SCC should not be done. In the other hand, negative phenotypic correlations demonstrated that as the SCC increased, the milk production decreased.

Purification and properties of buffalo (Bubalus bubalis) erythrocyte hexokinase

Buffalo erythrocytes contain one isozyme of hexokinase that apparently lacks microheterogeneity as shown by chromatographic properties. A single protein band was detected by means of Western blotting using an antibody raised in rabbits against homogeneous rat brain hexokinase I. The native protein has a molecular weight of 200,000 +/- 2880 by gel filtration. Partial purification of erythrocyte hexokinase by a combination of several procedures, including affinity chromatography, which was previously applied successfully to the purifica tion of other mammalian type I hexokinases, produced a partially purified enzyme that showed several contami nants after SDS-polyacrylamide gel electrophoresis. The affinity of buffalo erythrocyte hexokinase for glucose (K-m = 0.012 +/- 0.001 mM) is lower than most other mammal hexokinases type I. It phosphorylates other sugars, with considerably higher K-m values. This isozyme is able to use MgATP but does not use MgGTP, MgCTP or MgUTP. We used inhibition patterns, obtained with products to elucidate enzyme sequential mechanisms. Our results are clearly in agreement with a random sequential mechanism and in disagreement with an ordered sequential mechanism with either glucose or ATP as the obligatory first substrates. The ADP inhibition was of mixed type with both ATP and glucose as substrates. (C) 1997 Elsevier B.V.

Using Mastitis Records and Somatic Cell Count Data

On-farm records are essential for managing mastitis in dairy herds. Mastitis records are a useful tool for caring for an individual cow, to monitor compliance of farm personnel working with groups of animals, to understand the epidemiology of mastitis in the herd, to ensure responsible drug utilization, and to document accountability in care of the cow. Herds have become larger and more people are involved with individual animal care. This article describes a records plan that can be used to monitor mastitis at the herd level, aid in decision-making processes for individual cows, and improve drug use on dairy herds.

The relationship between dairy cow hygiene and somatic cell count in milk

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

Influence of hot environments on some blood variables of sheep

Thirty-two Polwarth sheep of ages up to 1 year were observed under temperatures varying from 10.5 to 46.5°C. The following blood cell counts were made: erythrocyte (RBC), leucocyte (WBC), eosinophil (EOS), neutrophil (NEU), lymphocyte (LYM) and monocyte (MON). Other traits measured were: haemoglobin (HB), haematocrit (HT), blood glucose (GLU) and serum protein (PROT). Multivariate analysis of variance was used and the results showed a significant (P<0.001) effect for the interaction of shearing and temperature treatment. Under temperatures >25°C, sheep presented a decrease of RBC, WBC, HB and HT, these differences being greater in the shorn than in the unshorn animals. Unshorn animals presented higher variations in EOS, NEU, LYM, MON and GLU. Blood glucose increased under high temperatures in the shorn animals (from 56.36±0.65 mg/100 ml to 60.52±0.69 mg/100 ml) as in the unshorn animals (from 54.72±0.74 mg/100 ml to 57.56±0.77 mg/100 ml). © 1992 International Society of Biometeorology.

Short communication: Resazurin reducing time as an indicator of Bradyrhizobium viable cell count

A method based on the use of resazurin (RSZ) is described to determine the number of viable Bradyrhizobium japonicum cells in culture medium. The observation of RSZ reduction can be done spectrophotometrically or visually. B. japonicum strains behaved differently when the reducing time was considered. This methodology can be used to determine the number of viable cells during the liquid culture stage of inoculant production.

Avaliação clínica e hematológica em bezerros Nelore infectados experimentalmente com isolados de Babesia bigemina das regiões Sudeste, Nordeste e Norte do Brasil

A comparative study was made regarding the clinical and hematological alterations caused by isolates of Babesia bigemina from southeastern, northeastern and northern Brazil in experimentally infected Nelore calves. Eighteen calves between 7 and 9 months of age, without antibodies against Babesia sp and raised free from ticks, were used. Three animals were previously inoculated with 2.0×109 parasitic erythrocytes (PE) for each stabilate. The other 15 calves were subdivided into three groups, with five animals each, that were subinoculated with 1.0×1010 PE of the respective isolates. The clinical and hematological alterations were evaluated by the determination of parasitaemia, haemogram, plasmatic fibrinogen, reticulocyte count, descriptive analysis of the bone marrow and erythrocytic osmotic fragility, for 30 days, totalizing seven moments of observation. The follow-up of the immunological response by the indirect fluorescent antibody test was carried out daily until the 10th day after inoculation (DAI) and after that, on the 15th, 20th, 25th and 30th DAI. A mild clinical manifestation of the disease was observed. The laboratory findings revealed low levels of parasitaemia; decrease of the erythrogram values; absence of reticulocytes, initial decrease in the total count of leukocytes, neutrophils and lymphocytes with a posterior elevation of the number of these cells; hypercellularity of the erythrocytic series and decrease of the myeloid: erythroid relation which was more accentuated between the 8th and 12th DAI, and an increase of the erythrocytic osmotic fragility in the groups inoculated with the Southeast and Northeast isolates. None of the three isolates of B. bigemina gave rise to the clinical characteristic form of the disease, although they induced an humoral immune response.

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.

Immune Response to the Rhodococcus equi infection in high and low antibody-producing mice (selection IV-A)

Rhodococcus equi is a Gram-positive, facultative intracellular bacterium which infects macrophages and causes rhodococcal pneumonia and enteritis in foals. Recently, this agent has been recognized as an opportunistic pathogen for immunocompromised humans. Several murine experimental models have been used to study R. equi infection. High (H IV-A) and Low (L IV-A) antibody (Ab)-producers mice were obtained by bi-directional genetic selections for their ability to produce antibodies against sheep and human erythrocytes (Selection IV-A). These lines maintain their phenotypes of high and low responders also for other antigens than those of selection (multispeciflc effect). A higher macrophage activity in L IV-A mice has been described for several intracellular infectious agents, which could be responsible for their intense macrophage antigens (Ag)-handling and low Ab production. Due to these differences, L IV-A mice were found to exhibit a better performance to trigger an effective immune response towards intracellular pathogens. The objective of this work was to characterize the immune response of Selection IV-A against R. equi. H IV-A and L IV-A mice were infected with 2.0 × 10 6 CFU of ATCC 33701 +R. equi by intravenous route. With regards to bacterial clearance and survival assays, L IV-A mice were more resistant than H IV-A mice to virulent R. equi. L IV-A mice presented a higher hydrogen peroxide (H 2O 2) and nitric oxide (NO) endogenous production by splenic macrophages than H IV-A mice. L IV-A expressed the most intense cellular response, available by the Delayed-Type Hypersensitivity (DTH) reaction, which activated macrophages and produced more H 2O 2 and NO. The three times higher specific antibodies titres in H IV-A indicated that Selection IV-A maintained the multispecific effect and the polygenic control of humoral and cellular responses also to R. equi.

Significado da presença de esquizócitos no sangue periférico de gestantes com pré-eclâmpsia

PURPOSE: to evaluate the significance of schizocytes presence in peripheral blood smear of pregnant women with pre-eclampsia, identifying and correlating them with other markers of hemolysis and of the disease severity. METHODS: Seventh six glass slides of peripheral blood smear of pregnant women with pre-eclampsia have been evaluated. After the smear, the slides have been stained with Leishman's dye and stored till they were examined with a Leica, model DLMB microscope, provided with the Qwin Lite 2.5 software that made it possible to record the images of selected fields in CD-ROM. Ten fields with approximately 100 erythrocytes were counted in each glass slide. Schizocytes (irregular fragment or helmet-shaped, bite-shaped or triangular) were considered as present, when their percentage was equal or higher than 0.2%, their presence being correlated with other hemolysis markers (hemoglobin, total bilirubin, lactic desidrogenasis and reticulocytes), pre-eclampsia markers (proteinuria and platelet number). The Statistical Package in Social Science for Windows (SPSS), 10.0 version has been used for statistical analysis, at p<0.05. RESULTS: schizocytes have been present in 31.6% of the pregnant women with pre-eclampsia. In most (75%) of the blood smears there have been three or four schizocytes. There has been no correlation between schizocyte presence and any other hemolysis marker, any pre-eclampsia marker or disease severity. CONCLUSIONS: schizocytes have been identified in a small number and in less than a third of the pregnant women with pre-eclampsia. There has been no correlation with other hemolysis marker parameters or with the disease severity. This way, the presence of schizocytes is not a marker of the clinical evolution of pre-eclampsia.

Eritrograma e medição dos eritrócitos de avestruzes (Struthio camelus) em Sãojosé do Rio Preto-SP, Brasil

Ostriches' erythrocytic parameters help on the diagnosis of specific pathologies and serve as basic knowledge for studies in comparative avian pathology. To obtain reference values of erythrocyte indices for ostriches (Struthio camelus) raised in a commercial system in Brazil and verify if there are differences between gender and age groups, 240 healthy from both sexes animals were bled. Heparinized blood samples were analyzed using standard techniques to determine the red blood cell (RBC) count, hemoglobin concentration, packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC) values, besides the red cell morphometry analysis using a computer software that calculates the greater and the smaller diameters of the erythrocytes. Prior to data analysis, ostriches were divided into three different age groups: from four to 13 months, from 13 to 23 months and from 23 to 30 months the. Younger ostriches presented lower erythrocyte indices than the older ones. The age group effect was only significant in females for the erythrocyte, hemoglobin, MCV, MCH, and MCHC variables. The female ostriches presented PCV, MCV, MCH, and MCHC values significantly higher than males in some age groups. It was observed that the erythrocytes of the female ostriches are more elongated and larger than males. It was concluded that erythrocytic parameters of ostriches in São José do Rio Preto-SP, Brazil are influenced by gender and age, highlighting the importance of consider besides these factors also the geoclimatic conditions to an adequate interpretation of the erythrogram.

Determinação da toxicidade do selênio por meio de análises hematológicas em Oreochromis niloticus

Selenium (Se) is described as an essential micronutrient and participates in different biological functions, as the antioxidant defense systems maintenance and regulation. However, when in high concentrations, Se may cause toxic effects as well as hematological changes in fish. The aim of the present study was to determine the toxicity of selenium in the form of sodium selenate (Na 2Se 6+O 4) in Oreochromis niloticus based on hematological parameters, after exposure to different concentrations (0.01, 0.14 and 1.4 mg Se 6+ L -1). The erythrocytic and leukocytic series were examined over 14 days at intervals of 0, 3, 5, 7,10 and 14 days. The erythrocytic series showed significant alterations in the first 7 days, including the control group. Neutrophils and monocytes showed variations in the first 3 days at a concentration of 1.40 mgSe 6+ L -1 characterizing an acute response. The total number of leukocytes was different in relation to time zero on all Se concentrations. The thrombocyte count also differed statistically from time zero and control in the first 3 days at 0.14 mgSe 6+ L -1. These results indicate that different concentrations induce an acute response with diminution of total leukocytes, neutrophilia, monocytosis and thrombocytosis.