Effects of 4-nonylphenol on blood cells of the African catfish Clarias gariepinus (Burchell, 1822)

In the present work, the destructive effects of the 4-nonylphenol on one of the most economically important Nile fishes, namely African catfish (Clarias gariepinus) were studied. Apoptosis, erythrocytes alterations, micronucleus test and blood parameters count were used as biological indicators to detect those effects. After exposure to sublethal concentrations of 4-nonylphenol (0, 0.05, 0.08 and 0.1 mg/l), apoptotic red blood cells with many malformations and micronucleated erythrocytes were recorded. Decrease in the blood parameters such as red blood cells (RBCs), hemoglobin (Hb), package cell volume (PCV), mean corpuscular hemoglobin concentration (MCHC), platelets, white blood cells (WBCs), lymphocytes, basophils, monocytes and increase in mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), neutrophils, eosinophils indicated the negative effects of 4-nonylphenol. It was concluded that, the 4-nonylphenol caused genotoxicity in erythrocytes with many malformations in shape and number indicated with other blood parameters.     

Seasonal variations in red deer (Cervus elaphus) hematology related to antler growth and biometrics measurements

The aim of the study was to relate seasonal hematology changes with the rest of physiological variations suffered by red deer, such as antler and biometrics cycle, and to assess the relationship between hematology and the effort performed in antler development. Blood samples were taken from 21 male red deer every 4 weeks during 18 months. Samples were analyzed for the main hematological parameters. Simultaneously, biometrics measurements were taken, such as antler length, body weight, body condition score, testicular diameter (TD), and thoracic and neck girth. All the blood cell types (erythrocytes, leukocytes, and platelets) showed seasonal variations, increasing as antler cleaning approached, as did hematocrit and hemoglobin. The final size of antlers was negatively related to leukocyte count, nonlymphoid leukocyte count, red cell distribution width, mean corpuscular hemoglobin, mean platelet volume, and TD, whereas it was positively related to body condition during antler growth. Huge seasonal variations in some hematological values have been found to be related to changes in antler and biometrics measurements. Since these variations are even greater than the caused by deer handling, they should be taken into account when evaluating hematology in deer populations.     

The effect of repeated acute mental stress on habituation and recovery responses in hemoconcentration and blood cells in healthy men

Acute mental stress elicits hemoconcentration and polycytosis. We investigated whether haematological response to repeated acute mental stress would habituate and be sustained 45 min and 105 min after stress. Twenty-four men underwent a 13-min stressor three times, one week apart; hematological variables were measured at week one and three. Hematocrit, hemoglobin, leukocytes, lymphocytes, erythrocytes, and thrombocytes all increased from rest to immediately post-stress (p's<.001). After 105 min of recovery, leukocytes and platelets both were higher, and hematocrit, hemoglobin, lymphocytes, and erythrocytes were all lower than at rest (p's<.001 to <.05). At all time points, hematocrit (p=.005) and erythrocytes (p=.006) were lower at week three than at week one. In contrast to an attenuation in systolic blood pressure increase from rest to immediately post-stress (p<.001), and in cortisol recovery from immediately post-stress to 45 min post-stress (p<.001), the magnitude of change in hemoconcentration and cell counts in stress and recovery experienced no habituation. Adjustment for stress-induced plasma volume shift altered findings: Elevated leukocytes post-stress persisted at 105 min (p<.001); any changes in lymphocytes became insignificant; erythrocytes decreased from rest to post-stress (p<.001) to increase again during recovery (p's<.05); platelets increased linearly between rest and 105 min of recovery (p=.005). We conclude that the magnitude of changes in hemoconcentration and blood cells during acute mental stress and recovery failed to habituate to stress repeats and, in part, sustained up to 105 min. Plasma volume shift accompanying stress affects the time course of stress polycytosis.     

Erythrocyte lysis in isotonic solution of ammonium chloride: theoretical modeling and experimental verification

A mathematical model of erythrocyte lysis in isotonic solution of ammonium chloride is presented in frames of a statistical approach. The model is used to evaluate several parameters of mature erythrocytes (volume, surface area, hemoglobin concentration, number of anionic exchangers on membrane, elasticity and critical tension of membrane) through their sphering and lysis measured by a scanning flow cytometer (SFC). SFC allows measuring the light-scattering pattern (indicatrix) of an individual cell over the angular range from 10° to 60°. Comparison of the experimentally measured and theoretically calculated light scattering patterns allows discrimination of spherical from non-spherical erythrocytes and evaluation of volume and hemoglobin concentration for individual spherical cells. Three different processes were applied for erythrocytes sphering: (1) colloid osmotic lysis in isotonic solution of ammonium chloride, (2) isovolumetric sphering in the presence of sodium dodecyl sulphate and albumin in neutrally buffered isotonic saline, and (3) osmotic fragility test in hypotonic media. For the hemolysis in ammonium chloride, the evolution of distributions of sphered erythrocytes on volume and hemoglobin content was monitored in real-time experiments. The analysis of experimental data was performed in the context of a statistical approach, taking into account that parameters of erythrocytes vary from cell to cell.     

Dynamics of hematologic parameters and of the erythrocyte deformability index at the juvenal period of rats and guinea pigs

In experiments on Wistar rats and guinea pigs, dynamics of blood hematological parameters and of erythrocyte deformability index (DI) was studied from birth to the 3-month age. In rats, significant changes were revealed in the picture of blood for the first month of postnatal ontogenesis. The number of erythrocytes and leukocytes sharply increased, the number of reticulocytes decreased, their ratio in the degree of maturity was changed. The mean erythrocyte volume and their acidic resistance also decreased. Prior to the 4-week age, DI increased, which was accompanied by an increase in the hemoglobin hydration and flattening of the erythrocyte shape. Then ID decreased and erythrocytes acquired the more spherical shape. After the critical period of 3-4 weeks the studied parameters reached plateau. In guinea pigs, at this period of ontogenesis the hematologic parameters had sufficiently stable values, except for a DI decrease from birth to 1.5 months, then it rose without reaching the initial values. Like in the case with rats, DI had a close correlation with the erythrocyte parameter of shape (O min).     

Dynamics of hematologic parameters and of the erythrocyte deformability index at the juvenal period of rats and guinea pigs

In experiments on Wistar rats and guinea pigs, dynamics of blood hematological parameters and of erythrocyte deformability index (DI) was studied from birth to the 3-month age. In rats, significant changes were revealed in the picture of blood for the first month of postnatal ontogenesis. The number of erythrocytes and leukocytes sharply increased, the number of reticulocytes decreased, their ratio in the degree of maturity was changed. The mean erythrocyte volume and their acidic resistance also decreased. Prior to the 4-week age, DI increased, which was accompanied by an increase in the hemoglobin hydration and flattening of the erythrocyte shape. Then DI decreased and erythrocytes acquired the more spherical shape. After the critical period of 3–4 weeks the studied parameters reached plateau. In guinea pigs, at this period of ontogenesis the hematologic parameters had sufficiently stable values, except for a DI decrease from birth to 1.5 months, then it rose without reaching the initial values. Like in the case with rats, DI had a close correlation with the erythrocyte parameter of shape (O_min).     

Comparative evaluation of morphofunctional organization of vertebrate nuclear blood cells

Morphometrical parameters, osmoregulatory possibilities, and the membrane reserve value of nuclear hemocytes (leukocytes and erythrocytes) were studied in the main classes of vertebrates by using method of hypoosmotic loads. It has been established that in the fish--mammals line in erythrocytes the absolute reserve of the plasmalemma decreases and the relative area of the cell surface increases. Evolution of leukocytes is accompanied by an increase of the membrane reserve and of the surface area due to a decrease of volume and to a rise of folding of the plasmalemma.     

Influence of pregnancy, lactation and environment on haematological profiles in Danish landrace dairy goats (Capra hircus) of different parity.

1. Haematological analysis was performed in young and adult non-pregnant, non-lactating, early and late pregnant, early mid- and late lactating Danish landrace goats from five herds. The purpose was to determine whether their levels are significantly different in these states and the effect of age and parity on the changes. 2. The haematocrit, haemoglobin concentration, number of erythrocytes and leukocytes were higher in 8-12 month old non-pregnant and 1-2 year old pregnant goats than in adult pregnant and lactating goats of over 2 years of age. Haematocrit, haemoglobin concentration and the number of erythrocytes decreased in late pregnancy and early lactation. 3. At the same time mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration slightly increased. 4. The changes were directly proportional to parity, more in pluriparous than in primiparous goats. 5. After parturition the former group of parameters increased and the latter decreased. 6. There was an increase in the number of total leukocytes close to and after parturition due to neutrophils and lymphocytes and it was more pronounced in the first lactation than in pluriparous goats. 7. There were significant differences in many parameters between goats from different herds (within similar physiological states).     

Circadian rhythms of blood components in dementia senilis

In eleven patients with dementia senilis circadian rhythms of hematocrit, hemoglobin, erythrocytes, leukocytes, serum iron and transferrin were studied by means of the cosinor method. An evident circadian rhythm was observed for all blood components, except leukocytes. There were no significant differences between patients with dementia senilis and healthy aged persons as far as hematocrit, hemoglobin, erythrocytes, and leukocytes are concerned. Acrophase for serum iron and transferrin occurs a little earlier and mesor is lower than in healthy aged persons.     

On the kinetics of erythroid cell differentiation in fetal mice. II. DNA and hemoglobin measurements of individual erythroblasts during gestation

Microspectrophotometric absorption measurements were used to determine the hemoglobin content of erythroid cells derived from the yolk sac during gestation of fetal C3H mice, from day 9 to day 15. Using the DNA content as a marker for the mitotic state between 2C and 4C phase, five successive cell generations and their mean hemoglobin contents were distinguished: 12 pg (pg, picogram = 10^?12 gm). 22.2 pg, 37 pg, 50 pg and 56 pg. In the final state, nucleated erythrocytes contained 98 ± 22 pg hemoglobin. Erythroid cells derived from the liver were measured on day 15 of fetal gestation. The hemoglobin content of proerythroblasts was below 0.3 pg. The two cell generations in the basophilic state had 0.6 pg and 1.7 pg respectively. Polychromatic erythroblasts yielded a hemoglobin content of 5.1 pg in the first cell generation and 7.5 pg in the second one. Orthochromatic erythroblasts contained 8 pg, reticulocytes 12 pg and mature erythrocytes 28 ± 7 pg hemoglobin. Calculations based on these data suggest that the rate of total hemoglobin synthesis is similar in both yolk sac and liver erythropoiesis. The difference between the final hemoglobin content in nucleated erythrocytes of yolk sac origin and that in hepatic erythrocytes can be explained by the different cell generation times.     

Relations between number and volume of erythrocytes and leukocytes in the human blood

Total concentration of erythrocytes and leukocytes, a relative contents of neutrophils, lymphocytes, monocytes, average volume of these types of cells, were determined in humans. Changes in the cell indices seem to be correlated and sex-dependent. Thus, erythrocytes and leukocytes are correlated in their contents and volume characteristics. A control mechanism seems to exist in humans, involving microcirculation in capillaries.     

Hematotoxicity and blood glutathione levels after cisplatin treatment of tumor-bearing mice

The involvement of glutathione, a major cellular antioxidant, in cisplatin-mediated development of various hematological changes in mice bearing ascites Dalton lymphoma tumor was investigated. With tumor growth, glutathione levels decreased in blood but increased in tumor cells. Cisplatin treatment of tumor-bearing mice caused a decrease in glutathione levels in blood, ascites supernatant, and tumor cells. Blood hemoglobin, erythrocytes, packed cell volume and leukocytes (eosinophils, basophils, and lymphocytes) were also decreased along with the development of various morphological abnormalities in erythrocytes (microcytes, macrocytes, echinocytes, acanthocytes, etc.) after cisplatin treatment. All these hematotoxic features were noted to be increased more when buthionine sulfoximine (a specific glutathione-depleting agent) was also given prior to cisplatin treatment. However, combination treatment of cysteine (precursor for glutathione synthesis) plus cisplatin resulted in an improvement in the glutathione levels and decrease in hematological toxicities. It is noted that the glutathione levels in blood and abnormalities in erythrocytes and other hematological parameters are inversely related in cisplatin-mediated cancer chemotherapy. It is suggested that blood glutathione may play an important role in the development of cisplatin-mediated hematological toxicity in the host. This revised version was published online in July 2006 with corrections to the Cover Date.     

Hematology and serum chemistry of harp (Phoca groenlandica) and hooded seals (Cystophora cristata) during the breeding season, in the Gulf of St. Lawrence, Canada

Standard hematologic and serum chemistry parameters were determined from 28 harp seals (Phoca groenlandica) and 20 hooded seals (Cystophora cristata) sampled from 6 March 2001 to 13 March 2001 during the breeding season. Whole blood was collected immediately postmortem from harp seal mother-pup pairs and from six hooded seal pups, and from live-captured adult hooded seals and three hooded seal pups; blood was analyzed within 24 hr at a local human hospital. A certified veterinary laboratory validated subsamples of whole blood and analyzed all serum chemistry parameters. Significant interlaboratory differences in mean values of packed cell volume (PCV) and mean cell volume (MCV) were found. Significant differences were found between samples from the five seal groups (adult male hooded seals, lactating female hooded seals, unweaned hooded seal pups; lactating female harp seals, and unweaned harp seal pups) for hematology and most serum chemistry parameters. In general, age-class influenced mean values of PCV, hemoglobin (HB), red blood cell (RBC) counts, MCV, mean cell hemoglobin (MCH), mean cell hemoglobin concentration (MCHC), and nucleated red blood cell (NRBC) counts per 100 leucocytes, but most age-related variations were species specific. Harp seal pups had significantly lower mean values of HB, PCV, MCH, and MCHC than did other seal groups, and significantly lower mean RBC counts than did hooded seal pups. Mean NRBC counts per 100 leukocytes were more than three times higher in harp seal pups than in hooded seal pups, but this difference was not statistically significant. Mean MCV were significantly lower in harp and hooded seal pups compared to those of adult harp and hooded seals. Differences in hemograms between pup species were likely because of the precocious development of hooded seal pups, which are weaned within 4 days, compared to 12 days for harp seal pups. Among adult seal groups, male hooded seals had significantly higher mean values of PCV and HB than did female harp and hooded seals, and significantly higher mean RBC counts than did adult female hooded seals. Among adult females, mean values of MCH and MCHC were statistically higher in hooded seals than in harp seals. Adult female harp and hooded seals did not differ significantly in other RBC parameters and mean leukocyte counts. Mean values of glucose, blood urea nitrogen, total bilirubin, alanine aminotransferase (ALT), alkaline phosphatase (ALP), total protein, and albumin showed species-specific variations between adults and pups. Except for ALP, few significant differences in mean enzyme activities of aspartate aminotransferase (AST), ALT, creatine kinase and gamma-glutamyltransferase were found between seal groups. Mean concentrations of electrolytes (calcium, phosphorus, sodium, potassium, chloride, magnesium, and total carbon dioxide) varied with age class, but variations in potassium and magnesium were species specific. Harp seal pups had significantly higher mean phosphorus and potassium levels compared to other seal groups.     

Comparative evaluation of morphofunctional organization of nucleated blood cells of vertebrate animals

Morphometrical parameters, osmoregulatory possibilities, and the value of membrane reserve of nucleated hemocytes (leukocytes and erythrocytes) of the main vertebrate animal classes were studied with use of the method of hypoosmotic loadings. It has been established that in erythrocytes, in the fish??mammal row, the absolute reserve of plasmalemma increases, while the relative area of the cell surface decreases. Evolution of leukocytes is accompanied by an increase of the membrane reserve and the surface area at the expense of a decrease of the volume and a rise of folding of the plasmalemma.     

Basic hematological values in some wild ruminants in captivity

Analyses of hematological parameters were carried out on eight axis deer (Cervus axis), 12 fallow deer (Cervus dama), 16 red deer (Cervus elaphus hippelaphus), three sambar (Cervus unicolor), nine Père David deer (Elaphurus davidianus), 20 European bison (Bison bonasus), seven nilgai (Boselaphus tragocamelus), eight mouflon (Ovis musimon), four white-bearded gnu (Connochaetes taurinus) and six barbary sheep (Ammotragus lervia). The following parameters were determined: packed cell volume, hemoglobin concentration, red blood cell number, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, leukocyte number and differential leukocyte count. Some degree of interspecific variability was found in the artiodactyls for these hematological parameters. An inverse relationship between mean corpuscular volume and red blood cell number is described, and specific values for these parameters are given for the four subfamilies.     

The effect of transportation stress on circulating corticosteroids, enzyme activities and hematological values in laboratory dogs.

Stress during transportation was monitored in truck-transported Beagle dogs "IVTK HD". Cortisol (F), corticosterone (B), red and white blood cell parameters, and enzyme activities in plasma were estimated. Plasma levels of cortisol and corticosterone were increased significantly during transportation and returned to basal values overnight post arrival. Hematocrit, hemoglobin, erythrocytes and leukocytes increased moderately. No obvious differences in enzyme activities were observed. Only alpha-HBDH values dropped significantly at the end of the 9.5 h transport period. This study confirms, although individual reaction is remarkably different, transportation as a potent stressor for Beagle dogs.     

Evaluation of an electrochemical model of erythrocyte pH buffering using 31P nuclear magnetic resonance data

When erythrocytes are suspended in a solution of known composition the resultant values of such basic cell parameters as volume and pH are difficult to predict. To facilitate such predictions, we developed a mathematical model describing the passive transmembrane distribution of permeant species; three simultaneous equations were produced. Certain essential data required for the model were determined experimentally; these included the pH dependence of the charge on the hemoglobin molecule and the variation of the osmotic coefficient of hemoglobin with cell volume. Finally, cells were added to various solutions, and then titrated to produce a wide pH range (pH 6-8). We measured the resultant cell volume, cellular and extracellular pH using both conventional and 31P NMR methods. The expected equilibrium values of these electrochemical parameters were also calculated by solving (numerically) the three model equations. The accuracy of the model simulations was evaluated by direct comparison of calculated and experimentally determined values.     

Involvement of leukocytes in the regulation of erythrocyte electrokinetic properties

Factor analysis was used to study the interrelations between the electrophoretic mobility of erythrocytes and some characteristics of blood leukocytes of healthy subjects and patients with arterial hypertension. In health, the stabilization of the mean mobility of erythrocytes in the electric field was shown to be independent of the changes in the blood leukogram (mainly, owing to the redistribution in the population of the cells with different properties). In disease, the pattern of this redistribution is altered, and white blood cells are involved in the regulation of the electrokinetic parameters of erythrocytes: the latter became sensitive to the total count of leukocytes and to the counts of stab neutrophils, monocytes, and eosinophils. The mechanisms of the involvement of leukocytes in the regulation of the electrokinetic properties of erythrocytes are discussed.     

Erythrocyte carnitine: a study of erythrocyte fractions isolated by discontinuous density gradient centrifugation

Erythrocyte fractions of varying density were isolated by discontinuous density gradient centrifugation of washed erythrocytes of five subjects (three adults and two cord bloods). Free and total carnitine concentrations were determined in each gradient fraction to compare the carnitine content of less dense with more dense erythrocytes. Erythrocyte, leukocyte, and reticulocyte counts and hemoglobin were measured on all fractions of each gradient. The density gradient studies showed that the highest proportion of reticulocytes were associated with the least dense gradient fractions of all five subjects. Linear regression analyses revealed significant positive correlations (r = 0.94 to 0.99, P less than 0.02 to P less than 0.001) between the number of reticulocytes per fraction and the total or free carnitine concentrations per fraction for all subjects. No correlation was found between free or total carnitine and hemoglobin, number of erythrocytes, or number of leukocytes per fraction. It appears that erythrocyte carnitine is localized in circulating reticulocytes which have mitochondria and carnitine-dependent fatty acid metabolism.     

Total body haematocrit iron kinetics and erythrocyte life span in pigeons (Columba livia)

1. The mean pigeon erythrocyte life span was found to be 17-25 days by Cr51-labeled erythrocytes and 21 +/- 3.4 days by iron kinetics. 2. Total red blood cell volume has been calculated by Cr51-labeled erythrocytes while total plasma volume was determined both by a dye method and iron kinetic data. From these results total blood volume and total body haematocrit were found to be 0.090 +/- 0.002 ml/g body wt and 36 +/- 4.3%, respectively. 3. Venous haematocrit, haemoglobin concentration, erythrocyte count, mean corpuscular haemoglobin concentration, plasma iron and red blood cell iron have also been measured. 4. A significant difference between total body and venous haematocrit and a short mean red blood cell life span, due to ageing and to random destruction of erythrocytes were shown. 5. The above observations are compared with analogous available data for human beings and their physiological significance is discussed.