Hematologic Reference Intervals Research Articles

Comparison of bovine hematology reference intervals from 1957 to 2006

The most commonly used bovine hematology reference intervals were published in 1965. We found the results from healthy cattle in 2001 differed from those in many ways. Discovery of the original laboratory book used to calculate the 1965 values gave us the opportunity to evaluate whether hematology values of healthy cattle have changed over time. The purpose of this study was to establish hematology reference intervals for Holstein cows, compare selected hematologic results with similar population data from 1957, and compare these reference intervals with those of other North American veterinary schools and published values. Reference intervals were developed in 2001 using clinically healthy, bovine leukemia virus-negative, mid-lactation Holstein cows. Selected parts of the hemograms and neutrophil:lymphocyte (N:L) ratio were compared with those from healthy, age-matched Holstein cows evaluated in 1957. Bovine reference intervals were solicited from clinical pathology laboratories in North American veterinary colleges and analyzed for population characteristics and method of analysis. Between 1957 and 2001, mean neutrophil counts increased significantly, whereas lymphocyte, monocyte, and eosinophil counts and hemoglobin concentration decreased significantly. Mean N:L ratio increased significantly to 1.17. Most surveyed laboratories were using the 1965 reference intervals. Two other institutions that had developed reference intervals after 2000 had results similar to ours. Continued use of older bovine hematology reference intervals could lead to misinterpretation of within-reference neutrophil counts as neutrophilia and under-recognition of neutropenia, eosinophilia, monocytosis, or lymphocytosis. Use of N:L>1 as evidence of inflammation should be discontinued or used with great caution.

Interspecific differences in the haematological reference intervals in Tupinambis genus from Argentina (Sauria–Teiidae)

In order to establish haematological reference intervals in the genus Tupinambis, blood samples were taken from the ventral coccygeal vein from 100 specimens of Tupinambis merianae and Tupinambis rufescens from Argentina. The samples were used to determine the haematocrit values, red blood cell count, total leukocyte and differential leukocyte count, thrombocyte count, mean corpuscular volume, haemoglobin concentration, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration and these were compared between both species. Statistically significant differences were observed as functions of the species (p > 0.001); T. merianae appeared to have higher red blood cell counts, haematocrit and haemoglobin than T. rufescens. Conversely, T. rufescens showed statistically high values in leukocyte and thrombocyte counts compared to the other species of the genus.

Breed‐specific variation of hematologic and biochemical analytes in healthy adult Bernese Mountain dogs

Hematology and serum biochemistry reference intervals in dogs may be affected by internal factors, such as breed and age, and external factors, such as the environment, diet, and lifestyle. In humans, it is well established that geographic origin and age may have an impact on reference intervals and, therefore, more specific reference intervals are sought for subpopulations. The objective of this study was to validate and transfer standard laboratory reference intervals for healthy Bernese Mountain dogs and to create new intervals for analytes where the established laboratory reference intervals were rejected. The procedure was performed using the human Clinical and Laboratory Standards Institute-approved model modified for veterinary use. Thirty-two dogs were included in the study using a direct a priori method, as recommended. While 23 of the standard laboratory reference intervals were readily validated, 7 of the analytes (eosinophils, MCHC, alkaline phosphatase [ALP], gamma-glutamyltransferase, total bilirubin, amylase, and cholesterol) required new reference intervals according to the standard. These were calculated using the robust method. In particular, the new reference range for ALP was wide compared with the established laboratory reference interval. No clinical causes were found for differences in the results of these analytes. We found significant differences in 7 hematologic and serum biochemical analytes for which a breed-specific variation appears to be the most plausible explanation. Breed-specific reference intervals for Bernese Mountain dogs will help avoid misinterpretation of laboratory results in the diagnostic process.

Blood Chemistry and Hematologic Values in Free-Living Nestlings of Montagu's Harriers (Circus pygargus) in a Natural Habitat

The establishment of reference intervals for hematologic and blood chemistry values in a given species is essential for subsequent diagnosis of disease at rehabilitation facilities and in captive breeding programs. In this study, blood samples were obtained from nestlings (18 males and 22 females) of a natural-vegetation breeding population of Montagu's harrier (Circus pygargus) in eastern Spain in order to establish reference intervals for blood values. This is the first study documenting hematologic and biochemical reference intervals for free-living Montagu's harriers. The studied population may reflect typical blood values for the species, since it is a free-living population that seems not to be constrained by food availability, as it is the population of Montagu's harriers with the highest productivity within Europe. Differences related to sex, year, clutch size, brood size, and number of fledglings were analyzed. Packed cell volume and hemoglobin were significantly lower from nests with five chicks compared to values from nests with fewer chicks, indicating that the cost of rearing more than four chicks in this species may be excessive. Total protein, calcium (Ca), and phosphorus (P) were significantly lower in the year with lowest mean rainfall, which correlates with the worst breeding success rate since the population has been monitored. No gender differences were found; this is likely a result of the prepubertal age of the animals sampled.

Development and application of biochemical and haematological reference intervals to identify unhealthy green sea turtles (Chelonia mydas)

Biochemical and haematological reference intervals (RIs) have been reported for sea turtles, but their value for ante-mortem disease diagnosis may be limited due to small sample sizes and outdated statistical analyses. In the present study, 290 green sea turtles (Chelonia mydas) were captured, clinically assessed and blood sampled. Of these, 211 were classified as 'clinically healthy' and 25 as 'clinically unhealthy'. RIs were estimated using data from the healthy turtles and compared with blood values from the unhealthy animals. All of the unhealthy animals had plasma biochemical and haematological values outside one or more RIs (albumin, 48% of unhealthy animals; alkaline phosphatase, 35%; aspartate transaminase, 13%; creatinine, 30%; globulin, 3%; glucose, 34%; lactic dehydrogenase, 26%; phosphorus, 22%; sodium, 13%; thrombocytes, 57%; and monocytes, 5%). Among small immature turtles, those with Chelonibia testudinaria plastron barnacle counts 20 were three times more likely to be unhealthy than those with no barnacles. In addition, small immature and mature turtles were more likely to be unhealthy than large immature turtles.

Hematological reference values of healthy Malaysian population

Health and disease can only be distinguished by accurate and reliable reference values of a particular laboratory test. It is now a proven fact that there is considerable variation in hematology reference intervals depending on the demographic and preanalytical variables. There are evidences that values provided by manufacturers do not have appropriate application for all populations. Moreover, reference ranges provided by different laboratory manuals and books also do not solve this problem. We are presenting here normal reference ranges of Malaysian population. These values were determined by using Sysmex XE-2100 and ACL 9000 hematology and coagulation analyzers. Results from this study showed that there were considerable differences in the reference values from manufacturers, western population or laboratory manuals compared with those from the local population.

Hematologic and serum biochemical reference intervals for free-ranging common bottlenose dolphins (Tursiops truncatus) and variation in the distributions of clinicopathologic values related to geographic sampling site

To develop robust reference intervals for hematologic and serum biochemical variables by use of data derived from free-ranging bottlenose dolphins (Tursiops truncatus) and examine potential variation in distributions of clinicopathologic values related to sampling sites' geographic locations. 255 free-ranging bottlenose dolphins. Data from samples collected during multiple bottlenose dolphin capture-release projects conducted at 4 southeastern US coastal locations in 2000 through 2006 were combined to determine reference intervals for 52 clinicopathologic variables. A nonparametric bootstrap approach was applied to estimate 95th percentiles and associated 90% confidence intervals; the need for partitioning by length and sex classes was determined by testing for differences in estimated thresholds with a bootstrap method. When appropriate, quantile regression was used to determine continuous functions for 95th percentiles dependent on length. The proportion of out-of-range samples for all clinicopathologic measurements was examined for each geographic site, and multivariate ANOVA was applied to further explore variation in leukocyte subgroups. A need for partitioning by length and sex classes was indicated for many clinicopathologic variables. For each geographic site, few significant deviations from expected number of out-of-range samples were detected. Although mean leukocyte counts did not vary among sites, differences in the mean counts for leukocyte subgroups were identified. Although differences in the centrality of distributions for some variables were detected, the 95th percentiles estimated from the pooled data were robust and applicable across geographic sites. The derived reference intervals provide critical information for conducting bottlenose dolphin population health studies.

Hematology and serum clinical chemistry reference intervals for free‐ranging Scandinavian gray wolves (Canis lupus)

Scandinavian free-ranging wolves (Canis lupus) are endangered, such that laboratory data to assess their health status is increasingly important. Although wolves have been studied for decades, most biological information comes from captive animals. The objective of the present study was to establish reference intervals for 30 clinical chemical and 8 hematologic analytes in Scandinavian free-ranging wolves. All wolves were tracked and chemically immobilized from a helicopter before examination and blood sampling in the winter of 7 consecutive years (1998-2004). Seventy-nine blood samples were collected from 57 gray wolves, including 24 juveniles (24 samples), 17 adult females (25 samples), and 16 adult males (30 samples). Whole blood and serum samples were stored at refrigeration temperature for 1-3 days before hematologic analyses and for 1-5 days before serum biochemical analyses. Reference intervals were calculated as 95% confidence intervals except for juveniles where the minimum and maximum values were used. Significant differences were observed between adult and juvenile wolves for RBC parameters, alkaline phosphatase and amylase activities, and total protein, albumin, gamma-globulins, cholesterol, creatinine, calcium, chloride, magnesium, phosphate, and sodium concentrations. Compared with published reference values for captive wolves, reference intervals for free-ranging wolves reflected exercise activity associated with capture (higher creatine kinase activity, higher glucose concentration), and differences in nutritional status (higher urea concentration).

CLSI-derived hematology and biochemistry reference intervals for healthy adults in eastern and southern Africa.

BackgroundClinical laboratory reference intervals have not been established in many African countries, and non-local intervals are commonly used in clinical trials to screen and monitor adverse events (AEs) among African participants. Using laboratory reference intervals derived from other populations excludes potential trial volunteers in Africa and makes AE assessment challenging. The objective of this study was to establish clinical laboratory reference intervals for 25 hematology, immunology and biochemistry values among healthy African adults typical of those who might join a clinical trial.Methods and FindingsEqual proportions of men and women were invited to participate in a cross sectional study at seven clinical centers (Kigali, Rwanda; Masaka and Entebbe, Uganda; two in Nairobi and one in Kilifi, Kenya; and Lusaka, Zambia). All laboratories used hematology, immunology and biochemistry analyzers validated by an independent clinical laboratory. Clinical and Laboratory Standards Institute guidelines were followed to create study consensus intervals. For comparison, AE grading criteria published by the U.S. National Institute of Allergy and Infectious Diseases Division of AIDS (DAIDS) and other U.S. reference intervals were used. 2,990 potential volunteers were screened, and 2,105 (1,083 men and 1,022 women) were included in the analysis. While some significant gender and regional differences were observed, creating consensus African study intervals from the complete data was possible for 18 of the 25 analytes. Compared to reference intervals from the U.S., we found lower hematocrit and hemoglobin levels, particularly among women, lower white blood cell and neutrophil counts, and lower amylase. Both genders had elevated eosinophil counts, immunoglobulin G, total and direct bilirubin, lactate dehydrogenase and creatine phosphokinase, the latter being more pronounced among women. When graded against U.S.-derived DAIDS AE grading criteria, we observed 774 (35.3%) volunteers with grade one or higher results; 314 (14.9%) had elevated total bilirubin, and 201 (9.6%) had low neutrophil counts. These otherwise healthy volunteers would be excluded or would require special exemption to participate in many clinical trials.ConclusionsTo accelerate clinical trials in Africa, and to improve their scientific validity, locally appropriate reference ranges should be used. This study provides ranges that will inform inclusion criteria and evaluation of adverse events for studies in these regions of Africa.

Haematological, biochemical and selected acute phase protein reference intervals for weaned female Merino lambs

Merino lambs are currently the subject of much research into the welfare aspects of mulesing and mulesing alternatives. Obtain haematology, biochemistry and acute phase protein reference intervals using modern methodologies for female Merino lambs. Blood was collected from 50, weaned, 9- to 16-week-old, female Merino lambs. Haematology and biochemistry panels were performed using routine automated methods. The acute phase proteins, fibrinogen, serum amyloid A and haptoglobin, were also measured using commercially available techniques. The reference intervals were determined to be the central 95% of results. Differences in the concentrations for some analytes were seen when compared with reported studies in sheep, but may be explained by the use of sheep of a different signalment, as well as different methodologies for analyte measurement. Overall, most analytes gave similar values to those previously reported in other studies. Notable exceptions were alkaline phosphatase, phosphate and globulins, for which the different results were often attributed to the younger age of the sheep in the present study, and platelets and creatine kinase, for which the elevated levels may have been a result of stress and muscle exertion associated with blood collection and husbandry practices. Established haematological, biochemical and acute phase protein reference intervals are necessary for the investigation of the systemic impact of mulesing and mulesing alternatives and for the investigation of systemic diseases affecting weaned, 9- to 16-week-old, female Merino lambs in general.

Evaluation of equine hemograms using the ADVIA 120 as compared with an impedance counter and manual differential count

The ADVIA 120 is an automated laser cell counter widely used in veterinary medicine. Although specific software for equine samples is available and validated, only a few reports have been published comparing the ADVIA 120 with other methods for equine hemogram evaluation. The purpose of this study was to compare the hematologic values and reference intervals obtained on the ADVIA 120 with those obtained on an impedance cell counter and manual differential counts in healthy horses. EDTA-anticoagulated blood samples were obtained from 114 clinically healthy horses of various breeds, both sexes, and 2-6 years of age. Samples were stored for up to 12 hours at 4 degrees C and then analyzed on the ADVIA 120 and the Hemat 8. A 100-cell to 200-cell differential leukocyte count was performed by 3 independent observers on May-Grünwald-Giemsa-stained smears. Intra-assay precision of the ADVIA 120 was determined by analyzing 5 replicates each of 10 of the blood samples. Results from the ADVIA were significantly higher than those from the impedance counter for RBC count, total WBC count, hemoglobin concentration, red cell distribution width, MCH, and MCHC, and significantly lower for HCT and platelet count. Significantly higher neutrophil and basophil counts and significantly lower lymphocyte counts were obtained with the ADVIA 120 compared with manual counts. Based on Passing-Bablok regression analysis, RBC and platelet counts were in good agreement between the 2 analyzers; a constant and proportional bias was present for other values. Coefficients of variation for erythrocyte parameters on the ADVIA were <1%, but were higher for platelet (6%), total WBC (2%), differential WBC (4%-30%), and reticulocyte (75%) counts. Results obtained with equine samples on the ADVIA 120 were comparable with those obtained on an impedance counter; reference intervals differed statistically but overlapped. The ADVIA had poor precision for reticulocyte and differential leukocyte counts such that the latter should always be verified on smears.

Normal blood cells reference intervals of healthy adults at the Gaza Strip—Palestine

Hematological parameters are affected by different factors that include age, sex, smoking, ethnicity, and environmental altitude. It has been justified that each population must establish its own normal reference intervals to be used in clinical assessments and interpretations. Hematological reference intervals for adults from the Gaza Strip-Palestine have never been addressed. Therefore, this study was designed and aimed at the establishment of normal blood cells reference intervals for healthy adults at the Gaza Strip-Palestine. This study involved 89,491 apparently healthy individuals (from both sexes and from the different governorates of the Gaza Strip) who were referred to the Thalassemia Central Laboratory during the period from September 2000 until February 2008. Complete blood counts were performed. Subjects were categorized into subgroups according to gender, smoking habit, and age (15-18, 19-45, and >45 years old). For each subgroup, descriptive and comparative statistical analysis was performed for hematological parameters. The results showed substantial differences between males and females, between smokers and nonsmokers, and between the different age groups. Moreover, reference intervals derived from our population are markedly shifted downward as compared with Western European and American populations. It was concluded that separate and region-specific reference intervals based on gender, smoking, and age for the Palestinian population at the Gaza Strip should be generalized for clinical laboratories and clinical practitioners, which could help in interpreting laboratory hematological tests more specifically, and potentially develop the quality of medical care provided to patients.

Hematologic characteristics of captive western barred bandicoots (Perameles bougainville) from Western Australia

The western barred bandicoot (Perameles bougainville) is an Australian marsupial species now considered endangered as a consequence of habitat destruction and predation. A recently discovered papillomatosis syndrome is hindering efforts to repopulate this species. Hematology reference intervals have been lacking for P bougainville, preventing optimal interpretation of hematology results from wart-affected and clinically normal animals. The purpose of this study was to establish hematology reference values and describe morphologic characteristics of blood cells of healthy western barred bandicoots. Fifty-nine whole blood samples were collected by jugular venipuncture into EDTA from 47 clinically healthy captive western barred bandicoots at 3 locations on the Western Australian mainland. A CBC was performed using an ADVIA-120 analyzer. Data were compared on the basis of geographic location, sex, age, and lactation status, and reference intervals were calculated. Blood cell morphology was evaluated using light microscopy, and transmission and scanning electron microscopy. Significant differences were found based on sex (RBC indices, fibrinogen), age (% polychromatophilic RBCs), and geographic location (RBC, neutrophil, and lymphocyte counts, MCHC, % polychromatophilic RBCs, fibrinogen). Combined reference intervals were calculated for hemoglobin concentration (122-165 g/L), HCT (0.36-0.49 L/L), and total WBC (2.9-14.9 x 10(9)/L), monocyte (0-0.6 x 10(9)/L), eosinophil (0-0.9 x 10(9)/L), and total plasma protein (47-63 g/L) concentrations. Leukocyte, erythrocyte, and platelet morphology were similar to those of other marsupial peramelid species. Nuclei in neutrophils, monocytes, and eosinophils occasionally had an annular configuration. Reference intervals and blood cell morphology obtained in this study will be useful for the evaluation of laboratory data from ill animals and assist with population health monitoring of western barred bandicoots.

Ethnic variation in certain hematological and biochemical reference intervals in a south Indian healthy adult population

Background We established the biochemical and hematological reference intervals among a south Indian healthy adult population attending an HIV referral centre in Chennai, southern India. Methods In a cross sectional study, 213 study subjects (129 male and 84 female) were studied between March and August 2005. All of the parameters were analyzed using standard hematological and biochemical techniques. Results Certain biochemical (viz. total bilirubin, alanine transaminase, albumin, creatinine, total protein, lipid profile, creatine phospokinase, uric acid and lactate) and hematological (mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration and lymphocyte levels) parameters presented higher upper limits. In addition, the upper limits of white blood cell count, platelet count, hematocrit, red blood cell count and hemoglobin level were low in comparison to the currently reported ranges. Conclusion Ethnic variation in reference intervals was observed in certain biochemical and hematological analytes in a south Indian adult population.

Haematological and biochemical reference intervals for farmed channel catfish

The reference intervals for biochemical variables and red blood cell indices of healthy intensively bred channel catfish Ictalurus punctatus were determined. The blood variables were determined using standardized clinical methods. The reference intervals (25th and 75th percentiles) were established using a non‐parametric method. Reference intervals for plasma glucose, serum total protein, sodium, potassium, calcium, magnesium, chloride concentration, primary and secondary red blood cell indices were established. The haematological and biochemical reference intervals established may allow important clinical decisions about channel catfish.

Hematologic and Biochemical Reference Intervals for Mountain Goats (Oreamnos americanus): Effects of Capture Conditions

We evaluated 90th percentile reference intervals for hematologic and biochemical analytes from 45 mountain goats (Oreamnos americanus) and determined the effects on these levels from sex, age, capture method, capture intensity, capture month, and region of the Washington Cascade Range. Mountain goats captured by helicopter darting had a lower percent neutrophils and higher percent lymphocytes than did those captured by darting from the ground. Helicopter darted mountain goats had higher levels of sodium, anion gap, osmolality, and hemolytic index and lower carbon dioxide than ground darted mountain goats. Pursuit intensity (distance and elevation gained) positively affected percent lymphocytes, and anion gap and negatively affected mean cell hemoglobin, percent neutrophils, and carbon dioxide. Alkaline phosphatase decreased with age. Hematocrit, mean cell volume, mean cell hemoglobin concentration, percent basophils, blood urea nitrogen, creatinine, total protein, globulin, albumin:globulin ratio, and magnesium exhibited temporal (month) or geographic (region) effects.

Hematology and serum biochemistry values of trapped, healthy, free‐ranging rock hyraxes (Procavia capensis) and their association with age, sex, and gestational status

The rock hyrax (Procavia capensis) is an herbivore prevalent from South Africa to Turkey, and a most common zoo animal. Although many studies of hyrax diseases and physiology are available, clinicopathologic data are limited. The purpose of this study was to establish comprehensive hematologic and biochemical reference intervals for trapped, apparently healthy, free-ranging rock hyraxes using modern laboratory methods and to assess differences related to sex, gestation, and age. Blood samples were obtained from 27 healthy, free-ranging hyraxes under anesthesia. Gender, body weight, and gestational status were recorded. Hematologic (n = 25) and serum biochemical (n = 22) analyses were performed using standard automated methodology. Data for male vs female, adult vs juvenile, and pregnant vs nonpregnant female hyraxes were compared using the Mann-Whitney U-test. Associations between variables were assessed using Pearson's or Spearman rank correlation tests. Significant age- and sex-related, but not gestation-related differences were observed in several variables. Serum alkaline phosphatase activity and phosphorus concentration were significantly higher in juveniles compared with adults. A unique type of monocyte comprised 1-3% of leukocytes in 4 hyraxes. Markedly high serum creatine kinase (CK) activity was observed in most hyraxes. The large number of animals and the availability of sex, age, and gestational data in this study will be useful to zoo and wildlife veterinarians working with rock hyraxes. High serum concentrations of betahydroxybutyric acid in the rock hyrax, compared with dogs, cats, and ruminants, may be related to its unique digestive system. High CK activity may have been the result of a capture myopathy-like syndrome. The unique monocytes in hyraxes resemble those of elephants and are a novel finding in this species.

Hematology and plasma chemistry reference intervals for cultured shortnose sturgeon (Acipenser brevirostrum)

The shortnose sturgeon, Acipenser brevirostrum, is an imperiled species distributed along the Atlantic coast of North America. Interest in replenishing wild stocks with hatchery-reared fish has created a need for accurate hematologic and biochemical reference intervals to evaluate the health of both fish raised in aquaculture systems and fish in the wild. The objective of this study was to generate hematologic and biochemistry reference intervals for healthy shortnose sturgeon. Blood samples were collected in heparinized tubes from 77 shortnose sturgeon raised in flow-through aquaculture systems. Whole blood and plasma samples were analyzed for hematologic and biochemical variables using standard techniques. Reference intervals were calculated as the central 95% (percentile) of data. Hematologic reference intervals (n = 46) were as follows: PCV 26-46%, hemoglobin 5.7-8.7 g/dL, MCV 307-520 fL, MCH 65.9-107.1 pg, MCHC 15-30 g/dL, plasma proteins (refractometry) 2.8-6.0 g/dL, RBC count 0.65-1.09 x 10(6)/microL, total WBC count 28,376-90,789/microL, small lymphocytes 9063-56,656/microL, large lymphocytes 2122-10,435/microL, neutrophils 3758-33,592/microL, monocytes 0-7137/microL, eosinophils 0-1544/microL, thrombocyte-like cells 6863-23,046/microL, thrombocytes 32,205-122,179/microL, and neutrophil:lymphocyte ratio 0.068-1.026. Plasma chemistry reference intervals (n = 77) were as follows: total protein 2.7-5.3 g/dL, albumin 0.8-1.7 g/dL, globulins 1.8-3.7 mg/dL, creatinine 0-1.4 mg/dL, total bilirubin 0-0.1 mg/dL, alkaline phosphatase 47-497 U/L, aspartate aminotransferase 90-311 U/L, sodium 124-141 mmol/L, potassium 2.9-3.7 mmol/L, chloride 106-121 mmol/L, calcium 6.6-12.1 mg/dL, magnesium 1.6-2.3 mg/dL, phosphorus 5.1-8.1 mg/dL, glucose 37-74 mg/dL, cholesterol 42-133 mg/dL, and osmolality 232-289 mOsm/kg. Reference values reported here will be useful for the early detection, identification, and monitoring of disease and sublethal conditions in cultured shortnose sturgeon.

Reference intervals for hematologic and biochemical constituents and protein electrophoretic fractions in captive common buzzards (Buteo buteo)

Increasing interest in wildlife care leads to the need for new tools to evaluate animal health. Laboratory investigations require reference intervals against which to compare the results obtained. For common buzzards, only a few studies have been performed to establish hematologic and biochemical reference intervals. The aim of this work was to develop reference values for routine hematologic and biochemical constituents and protein electrophoretic fractions and evaluate possible seasonal differences in values for healthy common buzzards. Heparinized blood samples were collected from 23 captive, clinically healthy common buzzards between February 2001 and June 2003. A CBC, routine biochemical analysis, and protein electrophoresis were performed. Data distribution was assessed and results from birds sampled in spring, summer, and winter were compared. Results from alternative methods for hemoglobin (Hgb; estimated as HCT / 3 vs spectrophotometry), total protein (biuret vs refractometry), and albumin (bromcresol green vs electrophoresis) concentrations also were compared. Reference intervals were calculated as 10-90th percentiles. In spring and summer, total WBC and heterophil counts, and urea, total protein, prealbumin, and beta- and gamma-globulins concentrations were significantly different from winter values. Results obtained by alternative methods for Hgb, total protein, and albumin concentrations were significantly different from those obtained by standard methods, although estimated and spectrophotometric Hgb values were significantly correlated. The reference values obtained in this study for hematologic and plasma biochemical constituents and their seasonal variation in healthy, captive common buzzards will be useful in the clinical evaluation of these birds in rehabilitation settings.

Hematology of the sandbar shark, Carcharhinus plumbeus: standardization of complete blood count techniques for elasmobranchs

Standardized hematologic methods and reference intervals have not been established for cartilaginous fishes (sharks, skates, and rays) despite the large number of animals displayed in zoos and aquariums worldwide. The focus of this study was to validate CBC methods for sandbar shark (Carcharhinus plumbeus) blood, based on criteria established in human medicine, for the following tests: RBC count, total WBC count, PCV, hemoglobin (Hgb) concentration, and WBC differential percentages. Replicate CBCs were performed using blood samples from 5 captive sandbar sharks. Three protocols for RBC and total WBC counts were compared, as were different centrifugation times for PCV determination, and 2 methods for Hgb concentration. Means, minimum and maximum values, and CVs were compared to CAP and CLIA performance guidelines for human tests. Total WBC counts in a diluent modified for elasmobranch blood, Hgb concentration by the cyanmethemoglobin method after removal of nuclei, and WBC differential percentages showed acceptable performance. PCV results were acceptable when tubes were centrifuged for at least 5 minutes. Total RBC counts by all 3 methods exceeded the acceptable error for manual counts of human cells. Standardized CBC tests can be used as health assessment tools for elasmobranchs. Total RBC counts should be viewed as estimates.