Hematologic Analytes Research Articles

The effect of subchronic oral exposure to zearalenone on hematologic and biochemical analytes, and the blood redox status of adult rabbit bucks.

Zearalenone (ZEN) is a mycoestrogen with a ubiquitous presence in animal feeds, which also has hematotoxic, hepatotoxic, nephrotoxic, and immunotoxic properties. However, there is a paucity of literature that discusses the effects of ZEN on rabbits. The aim of this study was to evaluate the effect of a prolonged, low-level (50μg ZEN/kg body weight) exposure on the clinicopathologic and redox status analytes of rabbit bucks. Ten adult bucks were included in the study. Each underwent a 7-week control period, followed by a 7-week exposure period. Water or ZEN solutions were daily administered orally (0.5mL) during the control and exposure periods, respectively. Blood samples were collected weekly for Complete Blood Counts, serum biochemical analyte and reactive oxygen metabolite (ROM) measurements. Data were analyzed using a mixed model, and the level of significance was set at a P of <0.05. During the ZEN exposure period, significant increases were noted in the red blood cell distribution width (RDW) and mean platelet volumes (MPVs), as well as in the white blood cell, monocyte, and eosinophil counts. Significant increases were observed in aspartate aminotransferase and total bilirubin, whereas urea, creatinine, glucose, total calcium, sodium, and potassium concentrations were significantly decreased. The ROM concentrations did not differ significantly between the control and ZEN exposure periods. Under the present experimental conditions, ZEN affected some of the clinicopathologic analytes of adult rabbit bucks; these changes were mostly indicative of mild hepatocellular damage and dysfunction, inflammatory and/or allergic responses, and renal tubular damage. A ZEN dose of 50μg/kg body weight did not seem to affect the blood redox status of bucks, as evaluated by the ROM concentrations.

Next-generation reference intervals for pediatric hematology.

Background Interpreting hematology analytes in children is challenging due to the extensive changes in hematopoiesis that accompany physiological development and lead to pronounced sex- and age-specific dynamics. Continuous percentile charts from birth to adulthood allow accurate consideration of these dynamics. However, the ethical and practical challenges unique to pediatric reference intervals have restricted the creation of such percentile charts, and limitations in current approaches to laboratory test result displays restrict their use when guiding clinical decisions. Methods We employed an improved data-driven approach to create percentile charts from laboratory data collected during patient care in 10 German centers (9,576,910 samples from 358,292 patients, 412,905-1,278,987 samples per analyte). We demonstrate visualization of hematology test results using percentile charts and z-scores (www.pedref.org/hematology) and assess the potential of percentiles and z-scores to support diagnosis of different hematological diseases. Results We created percentile charts for hemoglobin, hematocrit, red cell indices, red cell count, red cell distribution width, white cell count and platelet count in girls and boys from birth to 18 years of age. Comparison of pediatricians evaluating complex clinical scenarios using percentile charts versus conventional/tabular representations shows that percentile charts can enhance physician assessment in selected example cases. Age-specific percentiles and z-scores, compared with absolute test results, improve the identification of children with blood count abnormalities and the discrimination between different hematological diseases. Conclusions The provided reference intervals enable precise assessment of pediatric hematology test results. Representation of test results using percentiles and z-scores facilitates their interpretation and demonstrates the potential of digital approaches to improve clinical decision-making.

Hematologic and serum biochemical reference intervals for wild eastern quolls (Dasyurus viverrinus): Variation by age, sex, and season.

The eastern quoll (Dasyurus viverrinus) is an endangered carnivorous marsupial that has recently suffered significant population declines. Several small captive breeding populations have been established, with plans to translocate wild and captive individuals to areas of their former distribution. Accordingly, hematologic and serum biochemical reference intervals (RIs) established from wild eastern quoll populations are essential for monitoring the health and disease status of both captive and wild populations, and to evaluate the health of individuals before, during, and after translocation. We aimed to establish hematologic and serum biochemical RIs for wild eastern quolls, and examine the effects of age, sex, and season. We collected a total of 202 hematologic samples, 309 packed cell volume samples, and 335 serum biochemical samples from 168 individual quolls between May 2011 and November 2013. Species-level RIs were established, as well as RIs of groups separated by age (juvenile, adult) and sex (adult male, adult female) using nonparametric, robust, and parametric methods. Seasonal variation in age- and sex-specific reference values was also assessed. Strong age and seasonal variation were evident in many hematologic and serum biochemical analytes, with significant variation observed in serum biochemical analytes between the sexes. The observed age, sex, and seasonal variation reflect differences in the timing of growth and reproductive stressors, which interact with seasonal energetic demands. Our findings highlight the importance of using age-, sex-, and season-specific RIs for clinical evaluation of eastern quolls, as species-level RIs will inadvertently smooth and mask important seasonal fluctuations that reflect reproductive status at different times.

Age- and sex-specific reference intervals for hematologic analytes in Chinese children.

Reference intervals for pediatric laboratory tests need to be interpreted in the context of age- and sex-dependent dynamics. However, few reference intervals for healthy ethnic Han Chinese children have previously attempted to establish using large sample sizes. As such, there are no national hematological standards in China for pediatric reference intervals. We used a direct method to enroll a total of 2164 healthy 1- to 7-year-old children from Henan province. Hematological reference intervals were established by analyzing venous blood sample data on 17 hematologic analytes. The reference values for different ages and sexes were estimated using both parametric methods (mean±2SD) and nonparametric methods (2.5-97.5th percentiles). We provided reference intervals for 17 hematologic analytes including red blood count, hemoglobin concentration, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and red cell distribution width, white blood count and differential count as well as platelet count and related parameters. We established age- and sex-specific reference intervals that can provide more evidence-based guidance for the diagnosis and treatment for pediatric diseases. Our findings provide the basis for the next step in establishing national blood testing standards.

Hematologic reference intervals for the Italian Heavy Draft horse

Established breed-specific reference intervals (RIs) are important for monitoring the health status of horses. Breed-specific RIs have been evaluated in horses and other animal breeds, but there are no studies on breed-specific RIs for Italian Heavy Draft horse (IHDH). The aim of this study was to determine hematologic RIs for the IHDH living in central Italy. The comparison between different physiological status (pregnancy) and age (foals and adults) have been executed. Inclusion criteria were established for selection of subjects and for blood collection and handling. Blood samples were collected from 309 apparently healthy horses (mares, stallions and foals). Complete blood count (CBC) was performed on Sysmex-XT1800iV hematology at the Teaching Veterinary Hospital of the University of Perugia. RIs for IHDH were determined by using well-described, modern analytical and statistical methods as recommended by the CLSI and QALS guidelines. Hematologic RIs for the IHDH are reported. Differences between hot-blooded, cold-blooded, and warm-blooded horses are evident. Significant differences have been found for five analytes (WBC, MCHC, PLT, monocytes, basophils) between pregnant and non-pregnant mares and 12 analytes (RBC, Hgb, MCV, MCH, MCHC, RDW, PLT, WBC, lymphocytes, monocytes, eosinophils, basophils) between foals and adults. This is the first study reporting the hematologic RIs in the IHDH. The results suggest that breed-specific RIs should be used for IHDH due to the difference with published data regarding cold-blooded, hot-blooded, and warm-blooded breeds. Moreover, significant variations of hematological analytes have been found between subjects with different physiological status and age.

Indirect determination of hematology reference intervals in adult patients on Beckman Coulter UniCell DxH 800 and Abbott CELL-DYN Sapphire devices.

Background Conventional establishment of reference intervals for hematological analytes is challenging due to the need to recruit healthy persons. Indirect methods address this by deriving reference intervals from clinical laboratory databases which contain large datasets of both physiological and pathological test results. Methods We used the "Reference Limit Estimator" (RLE) to establish reference intervals for common hematology analytes in adults aged 18-60 years. One hundred and ninety-five samples from 44,519 patients, measured on two different devices in a tertiary care center were analyzed. We examined the influence of patient cohorts with an increasing proportion of abnormal test results, compared sample selection strategies, explored inter-device differences, and analyzed the stability of reference intervals in simulated datasets with varying overlap of pathological and physiological test results. Results Reference intervals for hemoglobin, hematocrit, red cell count and platelet count remained stable, even if large numbers of pathological samples were included. Reference intervals for red cell indices, red cell distribution width and leukocyte count were sufficiently stable, if patient cohorts with the highest fraction of pathological samples were excluded. In simulated datasets, estimated reference limits shifted, if the pathological dataset contributed more than 15%-20% of total samples and approximated the physiological distribution. Advanced sample selection techniques did not improve the algorithm's performance. Inter-device differences were small except for red cell distribution width. Conclusions The RLE is well-suited to create reference intervals from clinical laboratory databases even in the challenging setting of a adult tertiary care center. The procedure can be used as a complement for reference interval determination where conventional approaches are limited.

REFERENCE INTERVALS FOR ERYTHROCYTE SEDIMENTATION RATE, LACTATE, FIBRINOGEN, HEMATOLOGY, AND PLASMA PROTEIN ELECTROPHORESIS IN CLINICALLY HEALTHY CAPTIVE GOPHER TORTOISES ( GOPHERUS POLYPHEMUS).

Currently available tests for the diagnosis of inflammatory disease in reptiles are limited and poorly sensitive. However, a number of hematological and plasma biochemical analytes are validated in the diagnosis of inflammation in mammals. The objective of this study was to establish reference intervals for erythrocyte sedimentation rate, lactate, heat-precipitated fibrinogen, hematology, and plasma protein electrophoresis based on total protein by biuret method in 23 clinically healthy, captive gopher tortoises ( Gopherus polyphemus) after successful rehabilitation and to determine differences by age, sex, and season. In order to investigate biological differences, samples were collected in April, July, and November. There were no sex differences in any measured analyte; however, there were significant differences by age and season. Immature animals (<2 kg) had significantly higher total protein, albumin : globulin ratio, pre-albumin, albumin, and α-1 globulin than adults. Tortoises sampled in the spring season had significantly higher total solids (refractometer) and lower eosinophils compared with animals sampled in the summer. Further investigation is required to determine the clinical utility of these analytes in the diagnosis of inflammation in this species.

EFFECT OF ROUTINE HANDLING AND TRANSPORTATION ON BLOOD LEUKOCYTE CONCENTRATIONS AND PLASMA CORTICOSTERONE IN CAPTIVE HISPANIOLAN AMAZON PARROTS ( AMAZONA VENTRALIS).

Increased glucocorticoids cause a characteristic stress leukogram in mammalian taxa. It is assumed that avians exhibit a similar response, but to date, there have been no controlled studies to correlate serial endogenous corticosterone levels to hematologic values. An established flock of 18 Hispaniolan Amazon parrots ( Amazona ventralis) was used as a model in a crossover study. The treatment group was subjected to the stress of transport, restraint, and common clinical procedures with serial blood samples collected at 20-min intervals for hematology and corticosterone levels; the control group was sampled at the same intervals. Longitudinal data analysis was performed with linear mixed modeling. For all hematologic analytes, the baseline value had a significant positive effect on subsequent values (all P < 0.001). The white blood cell, heterophil, and eosinophil counts and heterophil to lymphocyte ratio increased over time in the treatment group, whereas it remained stable in the control group ( P = 0.016, P < 0.001, P < 0.001, P = 0.02, respectively, for the time*treatment effect). Lymphocyte absolute counts decreased over time, although not significantly; the decrease was significant for the relative lymphocyte count in the treatment group. Monocytes and basophils were not significantly altered. The treatment group had a higher mean corticosterone level overall than the control group by approximately 60% ( P = 0.008). The mean corticosterone level also increased over time in both groups by three- to fourfold ( P < 0.001) by 20 min then plateaued. These results demonstrate that some significant hematologic changes may arise with routine handling and transportation of birds and should be accounted for in hematologic interpretation of cell counts.

Uncertainty evaluation in clinical chemistry, immunoassay, hematology and coagulation analytes using only external quality assessment data.

Measurement uncertainty (MU) is a parameter associated with the result of a measurement that characterizes its dispersion. We report results for estimating MU following the application of a top-down procedure using only proficiency test data to establish uncertainty levels for various analytes. Data were obtained from 142 laboratories participating in the Beijing Center for Clinical Laboratory (BCCL) proficiency testing/external quality assessment (PT/EQA) schemes. The 24-month study included six selected PT shipments to obtain estimates for 50th percentile (median) and 90th percentile MUs and to compare those estimates to usual analytic goals. The number of laboratory participants varied for each trial. The expanded uncertainty (U) was calculated using a cover factor of k=2 for a confidence interval of 95%. All reproducibility, method and laboratory biases came from the PT/EQA data. The median U (k=2) ranged from 3.2% (plasma sodium, indirect ion selective electrode) to 32.8% (triglycerides, free glycerol blanking) for clinical chemistry analyte means from participants in the same method group. Immunoassay analyte median U results ranged from 11.3% (CA125 tumor marker, Roche) to 33.8% (prostate-specific antigen [PSA], Abbott). The range for median U was 3.5% (red blood cell [RBC], Abx) to 30.3% (fibrinogen [FBG], other) for hematology and coagulation analytes. The MUs for most analytes satisfied quality requirements. The use of PT/EQA data, when available, provides an effective means for estimating uncertainties associated with quantitative measurements. Thus, medical laboratories can calculate their own MUs. Proficiency testing organizers can provide participants with an additional MU estimate using only EQA data, which may be updated at the end of each survey.

Note on the unique physiologic state of loggerhead sea turtles (Caretta caretta) during nesting season as evidenced by a suite of health variables

Baseline hematologic and plasma biochemical analytes provide insight into wildlife health. In recent years, blood analytes have been used to infer foraging strategies and nutritional status of female sea turtles of different species during nesting season in an effort to determine if turtles at this life stage are capital breeders that forage little to none during nesting season. These changes in foraging during nesting have not been documented in loggerhead sea turtles (Caretta caretta). The objective of this study was to evaluate correlations between hematologic, plasma chemistry, immune function, and antioxidative analytes of female loggerhead turtles during nesting season to determine evidence of reduced foraging. We found that chloride tended to increase, while total protein, various plasma protein fractions (pre-albumin, alpha-1-globulins, beta-globulins, total globulins), total white blood cells, superoxide dismutase, reactive oxygen species, iron, and triglycerides decreased over the course of nesting season. These results suggest that loggerhead turtles rely on fat stores accumulated on foraging grounds to fuel their energetic costs during nesting. Our results also indicate alterations in hemodynamics, metabolism, and antioxidative capacity due to reduced foraging and high energy efforts of nesting, which lend further insight into the physiologic dynamics and catabolic state of sea turtles during nesting season.

Combined and breed‐specific RIs for hematologic, biochemical, and hormonal analytes in Chios and Florina adult rams

Clinical pathology investigations are essential to evaluate the health status and practice management of ruminants. Appropriate and accurate RIs are required for the interpretation of laboratory results. Despite substantial phenotypic differences and management-related disparities between male and female sheep, RIs in rams are sparse and limited by small reference populations and outdated methods. The purpose of this study was to establish RIs for hematologic, biochemical, and hormonal analytes in adult rams using controlled preanalytic and analytic procedures, and to investigate whether breed-dependent partitioning of RIs is required. Reference individuals were selected by a direct a priori method. Blood was collected from 128 healthy adult rams representing 2 Greek breeds (64 Chios and 64 Florina). Hematologic analysis was performed on the Advia 120, while differential WBC counts were manually performed on Giemsa-stained blood smears. Flexor E, AVL 9180 Electrolyte Analyzer, and Immulite 1000 were used for biochemical and hormonal analyses, respectively. Combined and breed-specific RIs were calculated using nonparametric and robust methods, respectively. Five outliers were detected based on laboratory results. Partitioning of RIs, based on both statistical and nonstatistical criteria, was recommended for some of the analytes. The RIs are in some agreement with those in the literature. The observed differences may be attributed to preanalytic and analytic factors. The RIs determined in this study can be used as a guide for the interpretation of laboratory results in rams and can potentially be adopted by veterinary laboratories, provided that similar equipment and reagents are used.

Sex-specific reference intervals of hematologic and biochemical analytes in Sprague-Dawley rats using the nonparametric rank percentile method.

BackgroundHematologic and biochemical analytes of Sprague-Dawley rats are commonly used to determine effects that were induced by treatment and to evaluate organ dysfunction in toxicological safety assessments, but reference intervals have not been well established for these analytes. Reference intervals as presently defined for these analytes in Sprague-Dawley rats have not used internationally recommended statistical method nor stratified by sex. Thus, we aimed to establish sex-specific reference intervals for hematologic and biochemical parameters in Sprague-Dawley rats according to Clinical and Laboratory Standards Institute C28-A3 and American Society for Veterinary Clinical Pathology guideline.MethodsHematology and biochemistry blood samples were collected from 500 healthy Sprague-Dawley rats (250 males and 250 females) in the control groups. We measured 24 hematologic analytes with the Sysmex XT-2100i analyzer, 9 biochemical analytes with the Olympus AU400 analyzer. We then determined statistically relevant sex partitions and calculated reference intervals, including corresponding 90% confidence intervals, using nonparametric rank percentile method.ResultsWe observed that most hematologic and biochemical analytes of Sprague-Dawley rats were significantly influenced by sex. Males had higher hemoglobin, hematocrit, red blood cell count, red cell distribution width, mean corpuscular volume, mean corpuscular hemoglobin, white blood cell count, neutrophils, lymphocytes, monocytes, percentage of neutrophils, percentage of monocytes, alanine aminotransferase, aspartate aminotransferase, and triglycerides compared to females. Females had higher mean corpuscular hemoglobin concentration, plateletcrit, platelet count, eosinophils, percentage of lymphocytes, percentage of eosinophils, creatinine, glucose, total cholesterol and urea compared to males. Sex partition was required for most hematologic and biochemical analytes in Sprague-Dawley rats. We established sex-specific reference intervals, including corresponding 90% confidence intervals, for Sprague-Dawley rats.ConclusionsUnderstanding the significant discrepancies in hematologic and biochemical analytes between male and female Sprague-Dawley rats provides important insight into physiological effects in test rats. Establishment of locally sex-specific reference intervals allows a more precise evaluation of animal quality and experimental results of Sprague-Dawley rats in our toxicology safety assessment.

Toxic elements and associations with hematology, plasma biochemistry, and protein electrophoresis in nesting loggerhead sea turtles (Caretta caretta) from Casey Key, Florida

Toxic elements (arsenic, cadmium, lead, mercury, selenium, thallium) are a group of contaminants that are known to elicit developmental, reproductive, general health, and immune system effects in reptiles, even at low concentrations. Reptiles, including marine turtles, are susceptible to accumulation of toxic elements due to their long life span, low metabolic rate, and highly efficient conversion of prey into biomass. The objectives of this study were to (1) document concentrations of arsenic, cadmium, lead, mercury, selenium, and thallium in whole blood and keratin from nesting loggerhead sea turtles (Caretta caretta) from Casey Key, Florida and document correlations thereof and (2) correlate whole blood toxic element concentrations to various hematological and plasma biochemistry analytes. Baselines for various hematological and plasma analytes and toxic elements in whole blood and keratin (i.e., scute) in nesting loggerheads are documented. Various correlations between the toxic elements and hematological and plasma biochemistry analytes were identified; however, the most intriguing were negative correlations between arsenic, cadmium, lead, and selenium with and α- and γ-globulins. Although various extrinsic and intrinsic variables such as dietary and feeding changes in nesting loggerheads need to be considered, this finding may suggest a link to altered humoral immunity. This study documents a suite of health variables of nesting loggerheads in correlation to contaminants and identifies the potential of toxic elements to impact the overall health of nesting turtles, thus presenting important implications for the conservation and management of this species.

Combining Nordtest method and bootstrap resampling for measurement uncertainty estimation of hematology analytes in a medical laboratory

BackgroundMeasurement uncertainty (MU) is a metrological concept, which can be used for objectively estimating the quality of test results in medical laboratories. The Nordtest guide recommends an approach that uses both internal quality control (IQC) and external quality assessment (EQA) data to evaluate the MU. Bootstrap resampling is employed to simulate the unknown distribution based on the mathematical statistics method using an existing small sample of data, where the aim is to transform the small sample into a large sample. However, there have been no reports of the utilization of this method in medical laboratories. Thus, this study applied the Nordtest guide approach based on bootstrap resampling for estimating the MU. MethodsWe estimated the MU for the white blood cell (WBC) count, red blood cell (RBC) count, hemoglobin (Hb), and platelets (Plt). First, we used 6months of IQC data and 12months of EQA data to calculate the MU according to the Nordtest method. Second, we combined the Nordtest method and bootstrap resampling with the quality control data and calculated the MU using MATLAB software. We then compared the MU results obtained using the two approaches. ResultsThe expanded uncertainty results determined for WBC, RBC, Hb, and Plt using the bootstrap resampling method were 4.39%, 2.43%, 3.04%, and 5.92%, respectively, and 4.38%, 2.42%, 3.02%, and 6.00% with the existing quality control data (U [k=2]). For WBC, RBC, Hb, and Plt, the differences between the results obtained using the two methods were lower than 1.33%. The expanded uncertainty values were all less than the target uncertainties. ConclusionsThe bootstrap resampling method allows the statistical analysis of the MU. Combining the Nordtest method and bootstrap resampling is considered a suitable alternative method for estimating the MU.

Reference intervals for hematological and biochemical parameters, acute phase proteins and markers of oxidation in Holstein dairy cows around 3 and 30 days after calving

The establishment of specific reference intervals (RIs) is advisable when different metabolic conditions in the patient population are present or when existing RIs are not suitable. Holstein dairy cows, at the start of lactation, experience a negative energy balance and an altered immune function that can lead to different levels of blood parameters compared with cows at peak lactation. The aim of this study was the determination of RIs for Holstein cows at 3±1 and 30±3days in milk. To this aim, 145 cows, from 4 herds, were sampled. A wide panel of hematological and biochemical analytes was determined, as well as the measurement of markers of inflammation/oxidation. RIs were generated following the American Society of Veterinary Clinical Pathology (ASVCP) guidelines and the effects of lactation period, parity, herd and day of sampling were determined. Data from 39 out of 52 analytes were significantly different according to lactation period. Data of 13 out of 52 analytes were not significantly different between the two groups. Some differences were also recorded between herds and parity groups but the adoption of specific RIs for these subgroups, however, has practical limitations. In conclusion the use of RIs specific for the lactation period is highly justified from both a statistical and a biological point of view. Pre-analytical factors associated with parity or management need to be considered in the evaluation of results of some analytes.

Impaired clinical utility of sequential patient GEM blood gas measurements associated with calibration schedule

BackgroundWithin- and/or between-instrument variation may falsely indicate patient trends or obscure real trends. We employ a methodology that transforms sequential intra-patient results into estimates of biologic and analytic variation. We previously derived realistic biologic variation (sb) of blood gas (BG) and hematology analytes. We extend this methodology to derive the imprecision of two GEM 4000 BG analyzers. MethodsA laboratory data repository provided arterial BG, electrolyte and metabolite results generated by two GEM 4000s on ICU patients in 2012–2013. We tabulated consecutive pairs of intra-patient results separated by increasing time interval between consecutive tests. The average between pair variations were regressed against time with the y-intercept representing the sum of the biologic variation and short term analytic variation: yo2=sb2+sa2. Using an equivalent equation for the Radiometer ABL, the imprecision of the two GEMs was calculated: saGEM=(yoGEM2−yoABL2+saABL2)1/2. This analysis was performed for nearly all measurements, regardless of time as well for values obtained over two 12h mutually exclusive periods, starting either at 2am or 2pm. ResultsRegression graphs were derived from 1800 patients' blood gas results with least 10,000 data pairs grouped into 2h intervals. The calculated saGEM exceed the directly measured saABL with many GEM sigma ratios of biologic variation/analytic variation being close to unity. All of the afternoon saGEM exceeded their morning counterparts with pH, pCO2, K and bicarbonate being statistically significant. ConclusionFor many analytes, the average analytical variation of tandem GEMs approximates the biologic variation, indicating impaired clinical usefulness of tandem sequential measurements. A significant component of this variation is due to increased variation of the GEMs between 2pm and 2am.

HEMATOLOGY AND PLASMA CHEMISTRY OF THE PLOUGHSHARE TORTOISE (ASTROCHELYS YNIPHORA) IN A CAPTIVE BREEDING PROGRAM.

Blood samples from 172 captive and 40 wild, healthy, juvenile and adult, ploughshare tortoises ( Astrochelys yniphora ) were analyzed to determine hematological and biochemical reference intervals. Hematological analytes included packed cell volume (PCV), white blood cell count (WBC), and WBC differential estimates. Biochemical analysis included total protein measured by photometry (TP) and by refractometry (TPr), albumin (ALB), creatine kinase (CK), aspartate aminotransferase (AST), glutamate dehydrogenase (GLDH), bile acids (BA), calcium (Ca), phosphorus (P), urea (UREA), and uric acid (UA). The jugular vein was identified as the preferred venipuncture site as subcarapacial vein venipuncture resulted in regular hemodilution. In due consideration of small sample sizes in some of the groups studied, adult tortoises had significantly higher plasma GLDH activity and TPr, TP, ALB, BA, and UREA concentrations and significantly lower AST activity and P concentration than juveniles. Captivity had a significant influence in some reference intervals, with captive adults presenting significantly higher WBC, and estimated counts of all white cell types as well as UREA and TPr than wild counterparts. Captive juveniles also showed significantly higher estimated monocyte and lower estimated eosinophil and basophil counts. Although these differences most likely reflect local environmental or dietary differences, without representing pathology or a deviation from the normal, they question the applicability of reference values from captive animals to wild animals and vice versa. Significant sex differences were only observed for PCV and UA. The reported reference intervals may serve as benchmarks for clinical assessment and conservation of this critically endangered species.

Effects of time and temperature on 48 routine chemistry, haematology and coagulation analytes in whole blood samples.

Background Phlebotomy for the purpose of blood analysis is often performed at remote locations, and samples are usually temporarily stored before transport to a central laboratory for analysis. The circumstances during storage and shipment may not meet the necessary requirements. If analysed anyway, false results may be generated. We therefore examined the influence of precentrifugation time and temperature of the most frequently requested tests in whole blood. Methods Healthy volunteers donated blood in which 48 analytes were tested. Routine chemistry was performed in lithium heparin tubes, haematology in ethylenediaminetetraacetic acid tubes, coagulation in citrate tubes and glucose in sodium fluoride tubes. One tube was measured directly. The others were kept at different temperatures (4, 8, 20 or 30℃) and stored for 4, 6, 8 or 24 h before analysis. Additionally, some analytes were examined at 12, 16, 24 and 28℃. The mean percentage deviation was compared with different decision levels, including the total allowable error. Results When using the total allowable error as an acceptable limit, most of the investigated analytes remained stable. However, bicarbonate is unstable at almost all tested time-points and temperatures. Calcium, lactate dehydrogenase, potassium and sodium are particularly affected at low temperatures, while phosphate is mainly affected at and above room temperature after 8 h. Conclusion We established the influence of time and temperature on a broad range of analytes, which may be applied to set the limits in transportation and storage of whole blood samples.

Reference intervals for biochemical and haematological analytes of juvenile captive-bred long-tailed macaques (Macaca fascicularis) from Mauritius with examination of the effects of individual characteristics using principal component analysis.

The Mauritian cynomolgus macaque (Macaca fascicularis) is widely used in biomedical research. Determining reference intervals for biochemical and haematological analytes provides an important tool for clinical diagnosis and pre-clinical research. Blood samples from 736 Mauritian long-tailed macaques were analysed to determine reference intervals of 13 biochemical and 10 haematological analytes. The need for partitioning the reference interval between males and females was determined. To examine the variation associated with age, body weight and sex on the analytes, our correlating, multivariate data set was first reduced using principal component analysis, and then the effect of these characteristics on factor scores was examined using GLM analysis. Partitioning of reference intervals, based on sex, was recommended for albumin, alkaline phosphatase and mean corpuscular haemoglobin. Sex significantly influenced the concentration of lymphocytes, granulocytes and white blood cells. These findings provide useful reference data for research involving Mauritian long-tailed macaques.

Reference Intervals of Hematology and Clinical Chemistry Analytes for 1-Year-Old Korean Children.

BackgroundReference intervals need to be established according to age. We established reference intervals of hematology and chemistry from community-based healthy 1-yr-old children and analyzed their iron status according to the feeding methods during the first six months after birth.MethodsA total of 887 children who received a medical check-up between 2010 and 2014 at Boramae Hospital (Seoul, Korea) were enrolled. A total of 534 children (247 boys and 287 girls) were enrolled as reference individuals after the exclusion of data obtained from children with suspected iron deficiency. Hematology and clinical chemistry analytes were measured, and the reference value of each analyte was estimated by using parametric (mean±2 SD) or nonparametric methods (2.5-97.5th percentile). Iron, total iron-binding capacity, and ferritin were measured, and transferrin saturation was calculated.ResultsAs there were no differences in the mean values between boys and girls, we established the reference intervals for 1-yr-old children regardless of sex. The analysis of serum iron status according to feeding methods during the first six months revealed higher iron, ferritin, and transferrin saturation levels in children exclusively or mainly fed formula than in children exclusively or mainly fed breast milk.ConclusionsWe established reference intervals of hematology and clinical chemistry analytes from community-based healthy children at one year of age. These reference intervals will be useful for interpreting results of medical check-ups at one year of age.