Mean Cellular Hemoglobin Concentration Research Articles

A-144 Three-wavelength Polarization Imaging Method for the Analysis of Individual Human Blood Cells With Dye-Enhanced Dispersion of Dilution Buffer

Abstract Background Performing basic hematology analysis in a near-patient setting enables more rapid clinical decision making in the process of diagnosis. Traditional blood analyzers in central labs use a complex combination of sensing principles and chemical modification of the sample to derive the necessary parameters for a complete blood count (CBC). To reduce cost and maintenance, varying approaches have been employed to determine a CBC with a single-step measurement technique. One common option is the use of optical microscopy to deliver highly reliable results, examples are fluorescence microscopy, digital holographic microscopy, or spatial light interference microscopy. However, most of these technologies lack a proper determination of cellular-level RBC parameters. Therefore, a non-interferometric, quantitative-phase measurement technique was developed to derive relevant CBC parameters from a single measurement, comparable to a standard lab analyzer. Methods The optical setup employed by this method enables the parallel acquisition of brightfield and differential interference contrast (DIC) images by polarization-sensitive cameras. By using three cameras with individual optical filters, quantitative phase and absorption information can be calculated, which enables the determination of mean cellular volume (MCV), mean cellular hemoglobin (MCH), and mean cellular hemoglobin concentration (MCHC) for red blood cells (RBCs). The whole-blood sample is diluted in a buffer containing a non-cell-penetrating dye to enhance optical refraction contrast and therefore precision in the dependent calculated quantities. The sample is then introduced to a glass flow cell, and the cells are allowed to sediment to the bottom for imaging. Machine learning is applied to the derived images to classify unstained native white blood cells (WBCs) to yield a 5-part differential, which distinguishes between neutrophil, monocyte, lymphocyte, eosinophil and basophil. The algorithm is trained by imaging FACS-validated, purified WBC subgroups. All blood samples are measured on an ADVIA® 2120 Hematology System for control. Results A one-step, optical CBC hematology system based on quantitative phase and absorption image analysis was developed. The correlations for MCV and MCHC compared to a standard lab CBC analyzer were 0.77 and 0.84. The accuracy for the 5-part differential WBC classification was approximately 0.8. To enable a visual inspection of the classification results, a pseudo-colored image of each cell was generated and uploaded in a review tool. Conclusion With a more-refined, less-expensive optical setup, a high-quality CBC analyzer suitable for use at the point of care can be derived with the presented method. A possible simplification could be the use of low-cost but high-resolution Fourier ptychography microscopy (FPM) technology. Imaging of blood samples on a single-cell level offers the potential for the analysis of even more morphological parameters, for example, disease-related changes in the cell membrane. Also, the adhesion behavior of the WBCs in the glass flow cell could indicate cell activation status and therefore give information about infection processes. *Trademark disclaimer: “ADVIA, and all associated marks are trademarks of Siemens Healthcare Diagnostics, Inc. or its affiliates. All other trademarks and brands are the property of their respective owners.”

Empowering frail older adults: multicomponent elastic-band exercises and BCAA supplementation unleash physical health and preserve haematological biomarkers.

The effectiveness of Branched Chain Amino Acids (BCAAs) supplementation on enhancing exercise performance in both young and older adults remains a topic of debate. Recent research suggests that BCAAs combined with regular exercise might have an impact on human erythropoiesis, blood dynamics, and iron homeostasis. Given the increasing longevity of the global population, it is crucial to investigate the potential benefits of BCAA supplementation and regular exercise as non-pharmacological interventions for improving the overall health of frail older adults. To assess the influence of a 40-week multicomponent exercise intervention (MEP) combined BCCA supplementation on the haematological indicators of frail older adults (83-93 years old) residing in nursing homes. A prospective, naturalistic, controlled clinical trial employing an intervention-washout-intervention was conducted for this purpose. The study included four experimental groups: MEP plus BCAA supplementation (MEP + BCAA, n = 8), MEP only (n = 7), BCAA supplementation only (n = 7), and control group non exercising (CG, n = 13). Fried's physical frailty (PF) protocol was employed to stratify the participants. Additionally, the assessment included the evaluation of nutritional status, comorbidities, and anthropometric measurements. Among the several haematological markers examined, only mean cellular Haemoglobin Concentration (MCH) [F = 4.09; p < 0.03] and Mean Cell haemoglobin Concentration (MCHC) [F = 10, 323; p < 0,0001] showed significant effects of time group. Our findings demonstrate that a long-term intervention with BCAA plus MEP did not lead to significant alterations in the haematological profile. An 8-week withdrawal from interventions did not affect the frailty status in the MEP and MEP + BCAA groups, whereas the control group exhibited an increase in PF status. The findings, demonstrating the potential pro-immune effect and maintenance of MCH and MCHC levels, highlight the relevance of incorporating exercise and nutritional strategies to promote healthy aging. This study contributes to the achievement of the United Nations Sustainable Development Goals 3 (good health and well-being) and 10 (reduced Inequalities) for all.

Biological Variation and Reference Change Value of Routine Hematology Measurands in a Population of Managed Bottlenose Dolphins (Tursiops truncatus).

Hematological analyses are particularly useful in assessing a dolphin's health status. However, the creation of appropriate reference intervals for this species is difficult due to the low number of reference individuals. The implementation of individual reference intervals (iRIs) allows researchers to overcome this limitation and, moreover, also consider the within-individual variability. The aims of this study were (1) to evaluate the biological variations in some hematological measurands, including erythrocytes (RBC), hematocrit (Hct), mean cellular volume and hemoglobin content (MCV and MCHC, respectively), RBC distribution width (RDW), leukocytes (WBC), and platelets (PLT); and (2) to calculate the index of individuality (IoI) and reference change value (RCV), which enable the production of iRIs, in healthy managed bottlenose dolphins. Seven dolphins were included, and the results of six hematological exams were analyzed for each animal. Analytical imprecision (CVa), within-dolphin variation (CVi), and between-dolphins variations (CVg) were calculated, and the IoI and RCV were derived for each measurand. All the hematological measurands had intermediate IoI except WBC, for which Iol was low. The calculated RCV ranged from 10.33% (MCV) to 186.51% (WBC). The results reveal that the majority of hematological measurands have an intermediate level of individuality in dolphins, and thus the application of iRIs is appropriate. The calculated RCV can also be applied to other managed dolphins and could be useful in interpreting serial CBC exams.

Results of Randomized Comparative Efficacy Study of Iron Deficiency Anemia Management in Children with Iron III Hydroxide Polymaltose Complex and Iron Sulfate

Background. The guidelines on selection between iron preparations, iron sulfate (IS) and iron III hydroxide polymaltose complex (HPC), for iron deficiency anemia (IDA) management in pediatrics are contradictory. Objective. The aim of the study is to compare efficacy and safety of iron (III) HPC and IS for IDA treatment in children. Materials and methods. Randomization of children (aged from 1 month to 18 years) with IDA into 2 therapeutic groups was implemented in 2019-2020. on pediatric districts of children’s city outpatient's clinic: IS dosage of 3 mg/kg/day and iron (III) HPC dosage of 5 mg/kg/day. Hemogram monitoring was performed every 14 days. The efficacy was compared according to the rate of Hb level increase, erythrocytes indices, and serum ferritin (SF) level at the recorded moment of Hb normalization. For the safety — the rate of adverse effects. Results. 65 children with IDA were included in the study and randomized into 2 groups, the median age was 2.3 years (1st; 3rd quartile = 1.1; 4.3). Hb level increase was comparable in both groups (0.34 (0.23; 0.48) g/L/day and 0.24 (0.21; 0.30) g/L/day, р = 0.891). IS is more effective than iron (III) HPC in terms of average red blood cell volume by the time of Hb normalization (83.5 (80.0; 86.7) fl against 80.7 (79.0; 82.0) fl, р = 0.006), and mean cellular hemoglobin content (28.9 (SD = 2.0) pg against 27.4 (SD = 1.8) pg, р = 0.004). The timing of SF levels normalization did not differ. Adverse effects to iron III HPC treatment occurred 5.5 times more frequently than to IS (p = 0.0001). Conclusion. The efficacy of IS and iron III HPC at standard doses is comparable. The advantage in tolerability and recovery of erythrocytic indices justifies the feasibility of using IS preparations in the first line of treatment for children with IDA.

Effect of Feeding Albino Rats with Acha-Orange Fleshed Sweet Potato Blends on the Blood Glucose Level and Hematological Parameters

Aims: To investigate the health and the physiological status of farm animals.&#x0D; Study Design: We formulated seven (7) samples from ACHA and orange-fleshed sweet potato mixture with two (2) samples serving as control.&#x0D; Place and Duration of Study: Department of Food Science and Technology, Faculty of Agriculture and Life Sciences, Federal University Wukari between February 2022 and May 2022.&#x0D; Methodology: The orange fleshed sweet potato flour was mixed with ACHA flour separately at different proportions (100:0, 95:5, 90:10, 85:15, 80:20, 75:25 and 0:100) while one hundred percent (100%) acha flour and orange-fleshed sweet potato was used as the control. Bone meal (10%), blood meal (10%) and salt (5%) were added to each sample. The albino rats were kept in a cage in five (5) segments, and the blends were fed to the rats for 21 days and 100g of the formulated feed were weighed into plates in each segment. The daily weight of the albino rats was measured. Evaluated bioassay parameters including feed and water intake, and body weight. Hematological properties including hemoglobin, red blood cell, white blood cell, mean cellular volume, mean cellular hemoglobin, and mean cellular hemoglobin concentration of the albino rats were also measured.&#x0D; Results: The result of feed intake/weight gain revealed that the rats fed with 0% OFSP showed the highest values for 21 days. The kidney, liver and heart weight of the albino rats decreased from1.23 to 1.05, 5.65 to 2.65 and 1.28 to 0.23g, respectively, while the carcass increased from 104.55 to 111.55g upon OFSP substitution.&#x0D; Conclusion: The result of this study revealed that locally available food commodities such as ACHA and orange fleshed sweet potato can be utilized to produce a protein-rich complementary food capable of combating malnutrition among children. A protein and carbohydrate-rich weaning food comparable to commercial weaning food (cerelac) can be strategically formulated from the blends of ACHA and orange-fleshed sweet potato (75:25).

Iron Status Is Inversely Correlated with Circulating Red Cells in Patients with Sickle Cell Anemia

▪Introduction: Sickle cell anemia (SCA, HbSS) arises from the presence of sickle hemoglobin (HbS, a consequence of a single base mutation in HBB gene). Upon deoxygenation, deoxy-HbS polymerizes with detrimental effects on shape, function, and lifespan of the erythrocytes. Because the delay time of sickling is inversely proportional to the mean cellular hemoglobin concentration (MCHC), decreasing the concentration of HbS is an attractive therapeutic approach. We recently demonstrated that dietary iron restriction decreases MCHC with a concomitant improvement in hematologic parameters and a decreased propensity to sickle in the Townes murine model of sickle cell anemia. Data describing the relationship between iron status, MCHC, and hematologic parameters in SCA patients are limited to isolated case series that do not account for alpha thalassemia, a known modifier of SCA severity. Based on our observations in mice, we hypothesize that decreased iron availability is associated with improved hematologic parameters in SCA patients in the absence of alpha thalassemia. We investigated the relationship between MCHC and red cell count, as well as transferrin saturation and red cell count, in healthy volunteers and sickle cell patients without alpha thalassemia.Methods: A database consisting of 795 patients with sickle cell disease (all genotypes) attending the National Institutes of Health between 2001 and 2015 was queried (see table). Data were filtered for sickle genotype, whether on hydroxyurea therapy, transfusion status (HbA), alpha thalassemia genotype, and relevant hematologic and/or iron panels obtained on the same visit. Data from healthy volunteers were obtained from the National Institutes of Health Biomedical Translational Research Information System (BTRIS) database filtering for those without detectable HbS that had hematologic and iron panel data obtained on the same visit. Correlations were analyzed by the Pearson correlation coefficient, r. p-values < 0.05 were considered statistically significant.Results: Analyses were performed on patients with HbSS without detectable HbA that were not taking hydroxyurea and did not have alpha thalassemia. Thirty-nine of these patients had MCHC and red cell counts obtained on the same visit. Data indicate that MCHC is inversely correlated with red cell count in these patients (r=-0.423; p = 0.007). Importantly, no significant relationship between MCHC and red cell count was observed in healthy volunteers (n=40; r=-0.125; p =0.438). Transferrin saturation is directly correlated with circulating red cells in healthy volunteers (n=65; r=0.359; p=0.0033). However, in patients with sickle cell disease transferrin saturation has an inverse relationship with red cell count (n=53; r=-0.3131; p=0.0225).Conclusions: Transferrin saturation and MCHC have are inversely correlated with circulating red cell counts in patients with sickle cell disease. These results support the concept that decreased iron availability improves hematologic parameters in sickle cell disease. Future studies will focus on pharmacologic means to achieve this end. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Impact of Heat Stress on Growth Performance and some Blood and Physiological Parameters of Suckling Friesian Calves in Egypt

Forty newly born male and female Friesian calves including 20 calves during the winter season and 20 calves summer season 2020, were used to study the effect of heat stress on live body weight, body weight gain, feed intake, feed and economic efficiency during suckling period. Ambient temperature, relative humidity and temperature humidity index were markedly higher during summer in compression with winter season. Digestibility coefficients of all nutrients (DM, OM, CP, CF, EE and NFE) and feeding values (TDN and DCP) reduced significantly (P<0.05) for summer ration compared to winter ration. Ruminal pH value and ammonia nitrogen concentration were higher significantly (P<0.05), however total VFA concentration was lower significantly (P<0.05) for summer than winter season. Concentrations of serum total protein, globulin and glucose were higher significantly (P<0.05), however, albumin to globulin ratio, concentrations of urea and creatinine and activity of AST and ALT activity were lower significantly (P<0.05) for winter season compared to summer season. Hemoglobin (HGB) concentration and counts of white blood cells (WBC) and red blood cells (RBC) were declined significantly (P<0.05), however, haematocrit percentage (HCT), mean cellular volume (MCV), mean cellular hemoglobin (MCH) and mean cellular hemoglobin concentration (MCHC), were raised significantly (P<0.05) for summer season than those of winter season. Intake of TDN, CP and DCP were higher significantly (P<0.05) for winter season than those of summer season. Weaning weight, total weight gain and average daily gain were higher significantly (P<0.05) for winter season in comparison with summer season. The amounts of DM, TDN, CP and DCP per kg weight gain were lower significantly (P<0.05) for summer season compared to winter season. Feed cost per kg weight gain was higher significantly (P<0.05), however, output of ADG, net revenue and economic efficiency were higher significantly (P<0.05) for winter season than those of summer season. All physiological parameters such as rectum temperature (RT), skin temperature (ST), respiration rate (RR) and pulse rate (PR) were higher significantly (P < 0.05) during the summer compared to the winter season.

Phenotypic correlations between biologic markers and growth traits in Arbor acres broiler chicken strain

The study was conducted to determine the correlations between biologic markers,haematological indices and growth traits in Arbor Acres broiler strain. A total of 120 Arbor acres strain were used for the experiment which lasted for 56 days. The birds were raised in separate deep litter pens. Data collected include biologic markers: hemoglobin (Hb), packed cell volume (PCV), white blood cell (WBC), red blood cell (RBC), mean cell volume (MCV), mean cellular hemoglobin (MCH), mean cellular hemoglobin concentration (MCHC) and biochemical parameters include: blood glucose (GLU) and total serum protein(PRT). Growth traits include: body weight (BW), body length (BL), shank length (SL), keel length (KL), wing length (WL), breast width (BWD) and drumstick length (DSL). The result obtained showed that MCHC in week 4 it associated significantly with breast length(rp=0.989) and keel length (rp= -0.986), whereas in week 6, serum protein, MCH, PCV, WBC and RBC proved significant markers based on their significant association (P&lt;0.05;P&lt;0.01) with SL (rp=0.975), KL (rp=0.981), DSL (rp= -0.982; -0.989) and BL (rp= -0.991). In week 8, Hb, WBC, MCV, MCH and MCHC proved to be useful markers for selecting keel length, breast length, body length, body length and thigh width in Arbor Acre. Highest correlation was observed between WBC and BL (rp= -0.991) and between MCH and breast length (rp= 0.991) in the 6 week. Due to the limited associations observed in week 4, it may be more reliable to apply the biologic markers in selection programs, starting at the 6 week for Arbor Acres broiler strains. This study advocates that highly polymorphic DNA-based markers which are more reliable should be attempted for improving body weight of broilers.

Use of the reticulocyte channel warmed to 41°C of the XN-9000 analyzer in samples with the presence of cold agglutinins.

ObjectivesThe purpose of this study was to compare data obtained from the reticulocyte channel (RET channel) heated to 41 °C with those obtained from impedance channel (I-Channel) at room temperature in the samples with the mean corpuscular hemoglobin concentration (MCHC) < 370 g/L and in samples with the MCHC > 370 g/L, in the presence of cold agglutinins.MethodsIn this study, 60 blood samples (group 1) with the MCHC < 370 g/L (without cold agglutinins) and 78 blood samples (group 2) with the MCHC > 370 g/L (with cold agglutinins) were used to compare the two analytical channels of the XN-9000 analyzer in different preanalytical conditions. The parameters evaluated in both groups were the following: red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), mean cell volume (MCV), RBC-most frequent volume (R-MFV), mean hemoglobin concentration (MCH) and mean cellular hemoglobin concentration (MCHC).ResultsThe results of this study showed an excellent correlation with both channels of the XN-9000 analyzer in samples with and without cold agglutinins, except for the MCHC. The bias between the values obtained in the I-channel and those obtained in the RET channel of both groups was insignificant and remained within the limits of acceptability, as reported by Ricos et al. for all considered parameters, except for MCHC.ConclusionsThe presence of cold agglutinins in blood samples can be detected by a spurious lowering of the RBC count and by a spurious increase in the MCHC. The RET channel represents a great opportunity to correct the RBC count in a rapid manner without preheating. However, neither methodology can completely solve the residual presence of cold agglutinins in all samples, despite the MCHC values being < 370 g/L.

A possible role for hepcidin in the detection of iron deficiency in severely anaemic HIV-infected patients in Malawi.

Iron deficiency is a treatable cause of severe anaemia in low-and-middle-income-countries (LMIC). Diagnosing it remains challenging as peripheral blood markers poorly reflect bone-marrow iron deficiency (BM-ID), especially in the context of HIV-infection. Severely anaemic (haemoglobin ≤70g/l) HIV-infected adults were recruited at Queen Elizabeth Central Hospital, Blantyre, Malawi. BM-ID was evaluated. Accuracy of blood markers (including hepcidin, mean corpuscular volume, mean cellular haemoglobin concentration, serum iron, serum ferritin, soluble transferrin receptor (sTfR), sTfR index, sTfR-ratio) to detect BM-ID was evaluated by ROC area under the curve (AUCROC). Seventy-three patients were enrolled and 35 (48.0%) had BM-ID. Although hepcidin and MCV performed best (AUCROC of 0.593 and 0.545 respectively) all markers performed poorly in identifying BM-ID (ROC<0.6). The AUCROC of hepcidin in males was 0.767 (sensitivity 80%, specificity 78%) and in women 0.490 (sensitivity 60%, specificity 61%). BM-ID deficiency was common in severely anaemic HIV-infected patients. It is an important and potential treatable contributor to severe anaemia but lack of definitive biomarkers makes it difficult to accurately assess iron status in these patients. Further investigation of the potential of hepcidin is needed, including exploration of the differences in hepcidin results between males and females.

Haematological reference intervals of wild southern Australian koalas (Phascolarctos cinereus).

Current haematology reference intervals (RIs) for koalas were developed in northern Australian koalas, using low numbers and/or individuals of unknown Chlamydia pecorum and koala retrovirus (KoRV) status. This study developed haematological RIs for wild, clinically healthy southern Australian koalas of known C. pecorum and KoRV infection status and investigated the effects of population, age and sex. Haematological RIs were determined for 138 clinically healthy South Australian koalas (Mount Lofty Ranges [MLR], n = 68; Kangaroo Island, n = 70) examined in April 2016 and February 2017, respectively. C. pecorum and KoRV status were determined by PCR. RIs for southern koala haematological parameters were established for all koalas based on the finding that there were limited differences in haematological values in koalas with subclinical C. pecorum or KoRV infections (P > 0.05), except KoRV-infected koalas had a lower haematocrit than noninfected koalas. MLR koalas had significantly lower erythrocyte mass and leucocyte counts than Kangaroo Island koalas. Young koalas had significantly lower haemoglobin, haematocrit and higher mean cellular haemoglobin concentration and lymphocyte counts than adult koalas. MLR male koalas had elevated erythrocyte, leucocyte and neutrophil counts compared with MLR females. The haematological RIs developed in this study are based on a large number of clinically healthy koalas, where subclinical C. pecorum and KoRV infections had no effect on haematological values and will be a valuable tool during clinical examination and disease investigation by veterinarians and researchers Australia-wide.

Iron Restriction Improves Markers of Disease Severity in the Townes Mouse Model of Sickle Cell Anemia

Introduction: Sickle cell anemia (SCA) is caused by mutations in β-globin that result in the production of the abnormal hemoglobin, HbS, with deleterious effects on erythrocyte shape and life span. Because the propensity of erythrocytes to sickle is inversely proportional to the concentration of HbS in the cell, decreasing the mean cellular hemoglobin concentration (MCHC) represents a potential therapeutic approach. Whereas iron restriction in healthy individuals does not alter MCHC, concomitant iron deficiency has been associated with decreased MCHC in SCA patients. Isolated case reports have linked iron restricted erythropoiesis with decreased hemolysis, increased red cell lifespan, and improvement in certain outcomes in SCA patients. We systematically examined the effects of iron restriction on erythropoietic outcomes in SCA utilizing the Townes murine model to investigate the hypothesis that mice with dietary iron deficiency will demonstrate a decreased MCHC, decreased erythrocyte sickling propensity, and improved anemia compared with mice on an iron sufficient diet. Methods: Townes SCA mice were weaned to diets containing either 20 ppm iron (low) or 48 ppm iron (sufficient) and maintained on those diets until sacrifice at 2 months of age. Blood was collected for complete blood count by submandibular or cardiac puncture. Spleen weight was normalized to body weight for calculation of the splenic index. Red cell deformability, defined by the elongation index (EI), and the oxygen pressure at which sickling occurs (point of sickling) during deoxygenation were characterized by oxygenscan ektacytometry using a laser optical rotational red cell analyzer (Mechatronics, The Netherlands). Results: SCA mice fed a 20 ppm low iron diet demonstrate a significant decrease in MCHC compared to SCA mice fed a 48 ppm iron sufficient diet (17.7+1.1 vs 22.7+5.5 g/dL; p &lt;0.05). Mice fed the low iron diet produced more circulating RBCs (5.98+1.03 vs 4.61+0.84 x 106 cells/uL; p &lt;0.05) and a higher hematocrit (39.4+7.1 vs 24.6+4.6%; p &lt;0.05) with a corresponding decrease in splenic index (0.074+0.14 vs 0.087+0.006 g/g body weight; p &lt;0.05) compared to mice fed the iron sufficient diet. RBCs from mice fed the low iron diet showed a significant decrease in the oxygen pressure at the point of sickling compared to RBCs from mice fed the iron sufficient diet (31.9+4.7 vs 40.6+4.6 mmHg; p &lt;0.01). The low iron diet also resulted in an overall improvement in deformability as evidenced by an increase in the EI minimum compared to the iron sufficient diet (0.25+.07 vs 0.12+.03 au; p &lt;0.01). No significant differences between groups were found in maximum deformability (EI max). The improved EI minimum translates to a decreased change in elongation (Delta EI), defined as EI max minus EI min, in RBCs from mice fed the low iron diet compared to mice fed the iron sufficient diet (0.11+.08 vs .23+.02 au; p &lt;0.01). Conclusions: Decreased dietary iron intake is associated with a decrease in the MCHC in Townes SCA mice. The lower MCHC is accompanied by an amelioration of anemia, decreased extramedullary erythropoiesis, improved RBC deformability and a shift in the initiation of sickling to lower oxygen pressures during deoxygenation. We speculate that iron restriction may decrease the clinical complications arising from both hemolysis and sickling in SCA. Future studies will utilize this model system to further characterize the effects of manipulating iron homeostasis on erythropoietic outcomes in SCA. Disclosures Fleming: Protagonist: Membership on an entity's Board of Directors or advisory committees; Silence Therapeutics: Consultancy; Ultragenyx: Consultancy.

European seabass (Dicentrarchus labrax) performance, health status, immune response and intestinal morphology after feeding a mixture of plant proteins-containing diets

The feasibility of reducing the fish meal (FM) content, as much as possible, in juvenile seabass diets by using a combination of four plant nutrients (plant mix, PX): soybean meal, SBM; soyprotein concentrate, SPC; corn gluten, CG and maize flour, MF was investigated . Seabass (mean initial body weight, 4.74±0.04 g) were fed with one of four isonitrogenous and isolipidic diets formulated to progressively reduce FM concentration of control diet (CTRL), by incorporation the PX, to be 30% (FM30 diet), 25% (FM25 diet), and 20% (FM20 diet) over a 13-week growth period. Fish were fed the experimental diets in triplicate and the resulting effects on growth performance, nutrient utilization, biochemical and amino acid composition, major blood constituents, immunity indicators and intestinal histology were evaluated. At the end of the trial, the FM25-fed fish presented growth, feed utilization, and survival rates comparable (P>0.05) to the CTRL fish. In addition, this diet did not negatively influenced major blood constituents, immunity indicators or intestinal histomorphology. Further dietary FM reduction to 20% (FM20 diet) and elevation of PX inclusion level (~ 59%) have caused a significant (P<0.05)decline in final fish weight, haemoglobin, erythrocyte, leukocyte and lymphocyte counts, elevated leukocytes phagocytic-activityand caused some abnormalities in the proximal intestine (PI) morphological features, but did not negatively impacted haematocrit value, mean erythrocyte cell volume (MCV), mean cellular haemoglobin content (MCH), mean cell haemoglobin concentration (MCHC), monocytes, eosinophils, serum total protein and lysozyme activity. Based on these results, the FM25 formulation (containing approximately 54% PX) can be considered the appropriate for reducing dietary FM down to 25% without compromising the performance, health, immunity or PI morphology of juveniles’ seabass.

Cryopreservation of red blood cells: Effect on rheologic properties and associated metabolic and nitric oxide related parameters

AimHigh glycerol cryopreservation of red blood cells (RBCs) reduces metabolic processes at ultralow temperatures but less is known regarding the effect of cryopreservation on RBC nitric oxide (NO) metabolism, haemorheological properties, structural behaviour and membrane fragility. MethodsBlood from ten healthy participants was sampled, glycerolized and stored at −80 °C (SB). Aliquots were thawed and further processed after 4, 8 and 12 weeks, respectively. At these time points, fresh blood (FB) was additionally sampled from each participant. FB/SB mixtures were prepared corresponding to transfusion of 1–3 blood bags. Additionally, mixtures were exposed to shear stress similar to that found in the circulation and deformability was measured to estimate possible behaviour of cryopreserved RBC in vivo. ResultsAgeing of RBC was reduced during cryopreservation. Markers for RBC metabolism (ATP, 2,3-DPG) were not altered but RBC sodium levels increased and potassium and calcium decreased, respectively. Mean cellular volume was higher and accordingly, mean cellular haemoglobin concentration was lower in SB. Deformability was altered during storage with less shear stress necessary to deform RBCs. Changes were also detectable in blood mixtures. Deformability remained unaltered in shear stress settings in FB and SB. RBC viscosity was reduced in SB. RBC-NOS content and phosphorylation sites as well as nitrite and RxNO levels seem not to be affected by the intervention. ConclusionCryopreservation maintains RBC metabolic function in vitro, but structure and function of cryopreserved RBC seems to be altered. Impact of these alterations in vivo seems to be less but needs further investigation.

Responses of the freshwater fish Cyprinus carpio exposed to different concentrations of butachlor and oxadiazon

The toxic effects of butachlor and oxadiazon, widely used herbicides in South Korea were evaluated in the common carp Cyprinus carpio. Fingerlings of C. carpio were exposed to 0.5, 5 and 50 µg L-1 concentrations of herbicides for 96 h under static bioassay method and hematological and biochemical parameters were assessed. At all concentrations of butachlor and oxadiazon, white blood cell (WBC), serum sodium (Na+), chloride (Cl-), glucose, cholesterol and triglyceride levels were significantly (P < 0.05) increased compared to that of their control groups. Mean cellular volume (MCV), mean cellular haemoglobin (MCH), potassium (K+) and alkaline phosphatase (ALP) levels were significantly increased in all the concentrations of oxadiazon exposed groups. Total protein, albumin and globulin levels were induced in butachlor treated fish. Both the herbicides caused a decrease in haemoglobin (Hb), hematocrit (Hct) and red blood cell (RBC) counts in all the concentrations. However, no significant changes were found in the mean cellular haemoglobin concentration (MCHC) levels. The importance of these parameters in monitoring of the aquatic ecosystem contaminated with pesticides is discussed in detail. Furthermore, chronic effects of these herbicides on these parameters along with other parameters such as hormonal and histopathological studies need to be investigated in the future studies.

Comparative study of haematological parameters of three sturgeon species in recirculating aquaculture system

The aim of this study was to evaluate and compare the haematological parameters of cultured Russian sturgeon (Acipenser gueldenstaedtii), stellate sturgeon (Acipenser stellatus) and Siberian sturgeon (Acipenser baerii) in recirculating aquaculture system. Total erythrocytes, thrombocytes, red blood cell distribution, monocytes, basophils as well as haemoglobin levels of stellate sturgeon differed significantly from that of the other two species (p&lt;0.05). Leukocyte and lymphocyte levels of Siberian sturgeon were significantly higher than in stellate sturgeon and Russian sturgeon (p&lt;0.05). Eosinophil percentages of Siberian sturgeon were significantly lower than stellate sturgeon and Russian sturgeon (p&lt;0.05). Haematocrit levels of stellate sturgeon differed from Siberian sturgeon (p&lt;0.05), but no differences between Siberian and Russian sturgeons were found. Neutrophil, mean cellular haemoglobin and mean cellular volume and mean cellular haemoglobin concentration levels did not differ significantly between the three sturgeon species (p&gt;0.05). The information generated may help in monitoring health status and to adopt better management strategies in culture systems.

Theoretical model for optical oximetry at the capillary level: exploring hemoglobin oxygen saturation through backscattering of single red blood cells.

Oxygen saturation ( sO 2 ) of red blood cells (RBCs) in capillaries can indirectly assess local tissue oxygenation and metabolic function. For example, the altered retinal oxygenation in diabetic retinopathy and local hypoxia during tumor development in cancer are reflected by abnormal sO 2 of local capillary networks. However, it is far from clear whether accurate label-free optical oximetry (i.e., measuring hemoglobin sO 2 ) is feasible from dispersed RBCs at the single capillary level. The sO 2 -dependent hemoglobin absorption contrast present in optical scattering signal is complicated by geometry-dependent scattering from RBCs. We present a numerical study of backscattering spectra from single RBCs based on the first-order Born approximation, considering practical factors: RBC orientations, size variation, and deformations. We show that the oscillatory spectral behavior of RBC geometries is smoothed by variations in cell size and orientation, resulting in clear sO 2 -dependent spectral contrast. In addition, this spectral contrast persists with different mean cellular hemoglobin content and different deformations of RBCs. This study shows for the first time the feasibility of, and provides a theoretical model for, label-free optical oximetry at the single capillary level using backscattering-based imaging modalities, challenging the popular view that such measurements are impossible at the single capillary level.

Hematological Parameters and Blood Cell Morphology of Male and Female Schizothorax (Racoma) davidi (Sauvage)

AbstractThe aim of this study was to obtain a basic knowledge of the hematology of Schizothorax (Racoma) davidi (Sauvage). The results showed that red blood cell and white blood cell counts as well as mean cellular hemoglobin concentration were significantly higher in females than in males, while mean cell volumes were significantly lower in females than in males. No significant differences were seen in other hematological parameters in relation with gender. Under light microscope, erythrocytes (mature and immature), leukocytes (neutrophils, lymphocytes, and monocytes), and thrombocytes were distinguished. Mature erythrocytes were ellipsoidal cells with oval, central nucleus. Neutrophils contained banded or segmented nuclei. Small lymphocytes had a very thin rim of basophilic cytoplasm. Monocytes often possessed vacuoles in their cytoplasm. Thrombocytes existed in several forms with relatively large nucleus surrounded by a pink‐stained or hyaline cytoplasm. The percentage of the different leukocytes revealed predominance of neutrophils and small lymphocytes, followed by monocytes and large lymphocytes. The neutrophil percentages of each stage showed that metamyelocyte accounted for the most, followed by banded neutrophils. This investigation could be used for evaluation of the health status of cultured male and female S. davidi and will provide early detection of clinical pathology.

Reticulocyte and erythrocyte hypochromia markers in detection of iron deficiency in adolescent female athletes.

The aim of this study was to analyse the effectiveness of new haematology parameters related to reticulocytes and mature red blood cells to differentiate pre latent and latent iron deficiency. The study included 219 female athletes (aged 15-20 years) representing volleyball, handball, cycling, canoeing, cross-country skiing, swimming and judo. To assess iron status the concentration of ferritin, soluble transferrin receptor (sTfR), iron and total iron binding capacity (TIBC) were determined in serum. In addition to blood morphology, the mean cellular haemoglobin content in erythrocytes (CH) and reticulocytes (CHr), mean cellular haemoglobin concentration in reticulocytes (CHCMr), the percentage of erythrocytes (HYPOm) and reticulocytes (HYPOr) with decreased cellular haemoglobin concentration, the percentage of erythrocytes (LowCHm) and reticulocytes (LowCHr) with decreased cellular haemoglobin content, and percentage of erythrocytes with decreased volume (MICROm) were determined. Subjects with ferritin <30 ng/ml were classified as having stage I (pre-latent) iron deficiency (ID). The second stage (latent ID) was diagnosed when low ferritin was accompanied by elevated sTfR and/or elevated TIBC values. The frequency of ID (without anaemia symptoms) was high, amounting to 60% (stage I in 45%, stage II in 15% of subjects). In subjects with stage I ID significant changes in haematological variables concerned mainly reticulocytes: CHCMr (p<.001), CHr (p<.05), LowCHr (p<.05), HYPOr (p<.001) in comparison to normal iron stores. In athletes with latent ID, there were also significant changes (p<.001) in many indices of mature red blood cells, i.e. haemoglobin concentration (Hb), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), CH, %LowCHm, as well as %MICROm (p<.01) in relation to the group without iron deficiency. The main finding of this study was that the diminished or exhausted iron stores had already caused changes in reticulocytes, and intensified iron deficiency (stage II) increased changes in both reticulocytes’ and erythrocytes’ hypochromia indices, while microcythaemia symptoms appeared later. This suggests that the markers of hypochromia relating especially to reticulocytes are useful for diagnosis of early ID in athletes with absence of an acute phase reaction.

Potential effects of low molecular weight phthalate esters (C16H22O4 and C12H14O4) on the freshwater fish Cyprinus carpio.

The aim of the present study is to assess the toxic effect of dibutyl phthalate (DBP) and diethyl phthalate (DEP) on the freshwater fish Cyprinus carpio. The median lethal concentrations of DBP and DEP for 96 h are found to be 35 and 53 mg L-1, respectively. Fish were exposed to 3.5 mg L-1 (Treatment I) and 1.75 mg L-1 (Treatment II) of DBP and 5.3 mg L-1 (Treatment I) and 2.65 mg L-1 (Treatment II) of DEP for a period of 35 days. The DBP and DEP exposed fish show a concentration based toxic effect on the selected parameters of this study. The hematological parameters, such as hemoglobin (Hb), hematocrit (Hct) and erythrocyte (RBC), were found to decrease in the DBP and DEP treated fish, whereas their leucocyte (WBC) count increased compared to that of the control groups. A biphasic response is noted in the erythrocyte indices, such as mean cellular volume (MCV), mean cellular hemoglobin (MCH) and mean cellular hemoglobin concentration (MCHC), throughout the study period. Exposure to DBP and DEP caused a significant (p < 0.05) decrease in sodium (Na+), potassium (K+), and chloride (Cl-) levels in the gill and brain of the fish throughout the study period when compared to that of their respective controls. The plasma protein level decreased in all the treatments, whereas the plasma glucose level significantly increased in the DBP and DEP exposed fish. Maximum inhibition of Na+/K+-ATPase activity was noticed in the gill and brain of the fish exposed to DBP and DEP. The cholinesterase (ChE) activity in the brain of the fish significantly decreased throughout the study period. A significant (p < 0.05) increase in glutamate oxaloacetate transaminase (GOT) and glutamic pyruvate transaminase (GPT) activity was noted in the fish exposed to both toxicants. The antioxidant enzymatic parameters such as superoxide dismutase (SOD) and catalase (CAT) activities were found to decrease in the gill and liver of the DBP and DEP treated fish, whereas a significant (p < 0.05) increase in lipid peroxidation (LPO) was observed. The above mentioned parameters could be used as potential biomarkers in clinical trials for the assessment of plasticizers. This study provides indispensable information towards future research on the effect of plasticizers on non-target organisms including humans.