Hematologic Reference Intervals Research Articles

Haematological and Biochemical Reference Intervals Towards a Proactive Health Monitoring Approach in Norwegian Atlantic Salmon Farming.

Haematological and plasma parameters are used in veterinary medicine as a diagnostic tool, allowing for the early detection of potential health and welfare issues. For the implementation of monitoring strategies, reference intervals (RIs) are needed to know when deviations in those parameters become clinically relevant. Here, haematological and plasma biochemical RIs of Norwegian Atlantic salmon (Salmo salar Linnaeus) are presented for 29 biomarkers. Two subgroups were defined from the reference population based on water type, referred to as pre-adult (freshwater, 166.8 ± 118.4 g, N = 979) and adult (seawater, 2655 ± 2883 g, N = 4941), collected at 93 fish farms along the Norwegian coast. While the influence of various pre-analytical variables on RIs is well documented, aquaculture practitioners cannot always control those variables in the field. Here, RIs were calculated from blood samples collected based on local pre-analytical practices and then further processed under standardised conditions (analytical phase). RIs are therefore considered highly relevant for the application in the field. Finally, when taking blood samples in the field, it is advised to centrifuge whole blood as soon as possible, while plasma parameters investigated were stable for 168 h at 4°C or 72 h at room temperature.

HEMATOLOGY AND BIOCHEMICAL REFERENCE INTERVALS FOR FREE-RANGING PRONGHORN (ANTILOCAPRA AMERICANA) IN WEST TEXAS.

Pronghorn (Antilocapra americana) are considered a keystone species of North American grasslands and an important economic source for many landowners in Texas. Pronghorn restoration projects routinely capture and translocate individuals from surplus populations to restoration areas. The objective of this study was to generate normal hematological and biochemical reference intervals (RI) for free-ranging pronghorn populations in Texas as a health monitoring tool for pronghorn restoration efforts. Blood samples were collected by jugular venipuncture and divided among an EDTA tube, serum separator tube, and a single blood smear on site. Complete blood counts and biochemical profiles were completed at the Texas Veterinary Medical Diagnostic Laboratory. In total, 417 individuals (41 males, 376 females) were included in the analysis. RI were determined by robust methods (R Studio) and mixed models' analysis of variance (SPSS 28) to examine differences in blood parameters due to fever, sex, age (adult versus yearling [<1 yr of age]), cell abnormalities, and pathogen exposure reported by the testing laboratory. Sex, age, and pathogen exposure affected mean blood values, but did not warrant development of separate RI by class. Bluetongue virus was identified in 46.8% (195/417) of pronghorns and epizootic hemorrhagic disease in 89.4% (194/217) of pronghorns; 84.8% (184/217) of the pronghorns tested positive for both diseases. This information provides baseline hematology and biochemical parameters to assess the health of free-ranging pronghorn and guide wildlife managers in decision-making for future translocations and restoration objectives.

Indirect Determination of Hematological Reference Intervals in Healthy Moroccan Adults Using Outpatient Data.

Reference intervals are essential for the interpretation of clinical laboratory tests and patient management. This study aims to determine age and gender reference intervals of complete blood count (CBC) in the Moroccan population by using the indirect approach. The study used data of ostensibly healthy adults collected retrospectively using the laboratory information system (LIS) of the Laboratory for Research and Medical Analysis of the Fraternal Royal Gendarmerie in Rabat (Morocco), between January 2018 and February 2020. The study included 5,898 men and 10,172 women ranging in age from 18 to 90 years. The lower and upper reference limits of CBC parameters were calculated using the nonparametric technique, as suggested by the Clinical and Laboratory Standards Institute (CLSI). All hematological parameters showed no clinically significant gender-related differences, except small differences in the values of hemoglobin (HB), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC). There were also no clinically significant agerelated differences for median values of all hematology analytes in both genders, except for platelet count (PLT) that continued to decline with increasing age in men and women, and Red blood cell count (RBC), Hematocrit (HCT), and hemoglobin (HB) that tended to increase with age but decrease in older age groups in men while they tended to increase with age in women. The indirect method can be used to establish reference intervals for CBC, with appropriate selection criteria and statistical tools. Our findings differed from the reference ranges provided in the textbook and also in other countries' reports.

Hematological reference intervals in newborn dromedary calves in the first week after birth: Age and sex-related variations

The establishment of hematological reference intervals (RIs) is an important tool to assess the health status of animals and to evaluate the impact of diseases at individual and population levels. Nowadays, specific RIs of hematological parameters in newborn dromedary camel calves at birth and during the first week after birth, are lacking. Therefore, RIs for the hematological variables from a complete blood cell count were established in 47 healthy newborn dromedary calves (18 females and 29 males). Blood samples were collected within 2 h after birth (d0), at 24 h (d1), at 3 (d3) and 7 days (d7) after birth, and analyzed within 24 h. The RIs were described based on the 95% confidence interval, and possible differences among mean values due to age (sampling time) and sex were investigated. Statistical analysis showed that age affected all the hematological variables except MCV, MCH, and MCHC, indicating that the adaptational process to the extrauterine life continues for several days after birth; sex affected most of the hematological variables, with higher RBC and PLT count, HGB, PCV, neutrophil population and neutrophil:lymphocyte ratio at d7 in females compared to males. These findings suggest possible sex-based differences in the physiological maturation mechanisms and deserves further investigations. To the best of the authors' knowledge, this is the first report of hematological RIs for newborn dromedary calves at birth up to 7 days of age; the RIs registered in the present study in newborns differ from those reported in adult dromedaries in literature, thus confirming the need for the adoption of separated reference ranges according to age also in the dromedary camel, as previously reported for other species.

Haematological reference intervals for pregnant Icelandic mares on pasture

BackgroundFew studies have been conducted on haematological reference intervals (RIs) in Icelandic horses. Reference intervals have been published for Icelandic horses in Austria and a preliminary study in Iceland compared haematological values in riding horses to published RIs for other breeds as well as Icelandic horses abroad. Haematological parameters can vary greatly due to factors such as breed, gender, age, reproductive status, and training, as well as feeding, prior exercise and management method. Icelandic broodmares are kept on pasture under supervision throughout the year, with haylage provided during the winter, and it is therefore of interest to establish haematological reference intervals for pregnant broodmares in Iceland. The purpose of this study was to establish haematological RIs specific to Icelandic broodmares in the first months of pregnancy, kept on pasture. Blood samples from 183 mares, stabilised in EDTA were analysed using IDEXX ProCyte Dx and total protein was analysed in serum samples from 157 of the 183 mares, using IDEXX Catalyst One analyser. The RIs were established using the guidelines of the American Society for Veterinary Clinical Pathology.ResultsThe RIs for red blood cell count, haematocrit and haemoglobin were higher in pasture-kept Icelandic mares in early pregnancy, most of which were lactating, than in pregnant mares of other breeds. This was also true for white blood cell count, as well as numbers of monocytes, eosinophils, and basophils, which in some instances might illustrate problems in the automated categorisation of some leukocytes.ConclusionsAs no RIs have been published for other pasture-kept Icelandic horses, future investigations should include other groups of pasture-kept Icelandic horses. Such an analysis might elucidate the effect of breed, management, and pregnancy on haematological values in pasture-kept Icelandic horses.

Hematological profile of healthy Thoroughbred foals from birth to one month of age in Trinidad, West Indies.

This study aimed to develop a sound database for the hematological reference intervals of thoroughbred foals in Trinidad, West Indies from birth to 1 month of age. 89 foals. Whole blood samples were taken from 89 foals throughout Trinidad at approximately 1 day, 1 week, and 1 month of age. These foals were examined to be classified as healthy or free from disease. Complete blood count (CBC), microscopic analysis of blood smears, and conventional PCR for Theileria equi and Babesia caballi were performed. Of the 89 foals, 67 were deemed healthy and suitable for establishing reference intervals. Foals in this study had lower mean hemoglobin and hematocrit values for all 3 times of sampling when compared to their North American counterparts. Age had a significant effect on hemoglobin, hematocrit, white blood cell (WBC), neutrophil, and platelet counts of the foals from birth to 1 month of age. Variations in reference intervals can occur due to differences in demographic, physiological, and environmental factors such as age, gender, breed, and geographical location. Given the changes in the hematological values over time, this study provides clinicians with valuable information that can be used to monitor the health status of newborn foals and detect disease conditions.

Reference Intervals of Haematological Parameters for Apparently Healthy Adults in Northeast Ethiopia.

Clinical laboratory reference intervals play a vital role in evaluating overall well-being, tracking the progression of diseases, and detecting potential harmful effects and complications. Despite evidence revealing disparities, many African nations currently rely on reference intervals for blood analysis obtained mainly from Western populations. This practice increases the risk of misidentifying and misdiagnosing healthy individuals. The aim of this study was to establish common hematological parameters reference intervals for healthy adults in Northeast Ethiopia. This community-based cross-sectional study consisted of 328 individuals who were presumed to be in good health. To assess their blood-related characteristics, blood samples were collected and analyzed using the advanced Dirui BF-6500 analyzer, along with serological testing. In accordance with guidelines provided by the Clinical and Laboratory Standards Institute, the study employed a non-parametric approach to calculate the medians and 95% confidence intervals. To explore potential variations between males and females, a statistical test known as the Mann-Whitney U-test was used to compare the reference intervals. The established reference intervals were: white blood cells 3.5-11.3×109/L; red blood cells 4.0-6.1×1012/L; hemoglobin 11.2-17.5g/dL; hematocrit 35.4-52.0%; MCV 77.9-93.8fl; MCH 24.7-32.0pg; MCHC 306-349g/L; RDW-CV 12.1-13.8% and platelet 131-391×109/L. The reference values of monocytes, eosinophils, red blood cells, hemoglobin, hematocrit and RDW-CV in males were higher than females, while females had significantly higher platelet counts compared to males. The reference intervals discovered differed from the reference intervals now in use, those mentioned in earlier research in Ethiopia or other African nations, as well as those conducted on Western populations. In the adult demographic of Northeast Ethiopia, specific reference intervals for commonly observed hematological parameters were established, tailored to the local community. Consequently, these reference intervals hold the potential to enhance informed decision-making within this population, by providing valuable guidance when interpreting laboratory test outcomes.

Haematological and biochemical reference intervals for apparently healthy working equids in The Gambia

Haematological and biochemical reference intervals for apparently healthy working equids in The Gambia

Establishment of Reference Intervals of Hematological Parameters and Evaluation of Sex and Age Effect in the Miranda Donkey.

The Miranda donkey is an autochthonous Portuguese breed that is considered endangered. Several studies have been carried out on this breed, but to the authors' best knowledge, no studies have been conducted on their clinical pathology. The aims of this study were to determine the hematological reference intervals (RIs) in healthy Miranda donkeys and to estimate the influence of age and sex. Blood samples from 75 clinically healthy animals were analyzed for 22 hematological parameters on the IDEXX ProCyte Dx, an automated hematology analyzer previously validated for the species. The RIs were estimated following the ASVCP guidelines with the Reference Value Advisor software. Regarding sex, no significant differences were found between groups. Regarding age, significant statistical differences (p < 0.05) were observed for red blood cells, red cell distribution width, white blood cells, lymphocytes, monocytes, platelets, plateletcrit (higher mean in young animals), mean corpuscular volume, mean corpuscular haemoglobin, neutrophils and eosinophils (higher mean in adults). The RIs described here can be used to evaluate and monitor the health status of animals and herds, as well as to guide diagnoses or select fit and healthy animals for reproduction, contributing to the preservation of the breed.

An Indian perspective for umbilical cord blood haematological parameters reference interval

BackgroundThe Haematological Reference Intervals (RIs) are prone to vary on the basis of various factors such as altitude, age, sex, socioeconomic status, etc. These values play a major role in laboratory data interpretation and determine the necessary clinical treatment. Currently, India has no well–established RI for cord blood haematological parameters of newborns. This study aims to establish these intervals from Mumbai, India.MethodA cross sectional study was conducted in a tertiary care hospital of India from October 2022 to December 2022 on healthy and term neonates having normal birth weight and born to healthy pregnant mothers. About 2 – 3 mL of cord blood was collected from the clamped cord into EDTA tubes from 127 term neonates. The samples were analysed in the haematology laboratory of the institute and the data was analysed. The upper and lower limits were determined using non-parametric method. The Mann–Whitney U test was used to compare the distribution of the parameters between sex of infant, modes of deliveries, maternal age and obstetric history. P value less than 0.05 was considered to declare statistical significance.ResultThe median values and 95% RI for umbilical cord blood haematological parameters of newborns were as follows: WBC = 12.35 [2.56–21.19] × 109/L, RBC = 4.34 [2.45–6.27] × 1012/L, HGB = 14.7 [8.08–21.44] g/dL, HCT = 48 [29–67]%, MCV = 109.6 [59.04–159.1] fL, MCH = 34.5 [30.54–37.79] pg, MCHC = 31.3 [29.87–32.75] %, PLT = 249 [16.97–479.46] × 109/L,LYM = 38 [17–62] %, NEU = 50 [26–74] %, EOS = 2.3 [0.1–4.8] %, MON = 7.3 [3.1–11.4], BAS = 0 [0–1]. This study found no statistically significant difference between sex of infants, except MCHC, and obstetric history. A significant difference was observed in WBC, EOS% and absolute NEU, LYM, MON and BAS by delivery type. A higher platelet count and absolute LYM was observed in the cord blood compared to venous blood.ConclusionsFor the first time, haematological reference intervals in cord blood were established for newborns in Mumbai, India. The values are applicable for newborns from this area. Larger study throughout the country is required.

Hematologic and Biochemical Reference Intervals of a Single Population of Captive Burmese Roofed Turtles (Batagur trivittata)

The Burmese roofed turtle (Batagur trivittata), endemic to Myanmar, is a critically endangered species. Little is known about their physiology and health. Therefore, establishing physiological reference intervals is essential for the implementation of conservation plans. This study establishes hematology and biochemistry reference intervals of a group of 23 captive Burmese roofed turtles. The animals were sampled during a four-month period, outside the breeding season. Most blood values did not differ significantly based on animal sex. However in females, lymphocyte count, and concentrations of glucose and calcium were relatively higher, and potassium lower compared to that of males. Better understanding of blood analytes will help strengthen ex-situ management and healthcare protocols, as well as improve ongoing conservation efforts for this important species.

Establishment and validation of haematological reference intervals for newborns aged 5 to 28 days in Nanjing, China

ABSTRACTBackground:China lacks a standard for reference intervals (RIs) of complete blood cell (CBC) counts in newborns. This study aimed to determine local haematological RIs for newborns.Methods:This prospective study was conducted from January 2020 to December 2020 in Nanjing Maternity and Child Health Care Hospital. We collected capillary blood specimens from 497 healthy newborns aged 5–28 days. We calculated the RIs as nonparametric 2.5th to 97.5th percentiles and 90% confidence intervals following the EP28-A3c guideline. We validated the RIs in another 20 specimens from healthy newborns.Results:The RIs for the 18 CBC parameters were: white blood cell count 7.17–15.69 × 109/L; monocytes# 0.52–1.66 × 109/L; Mono% 5.7–14.1; eosinophils# 0.16–1.08 × 109/L; Eos% 1.5–8.9; Lymphocytes# 4.04–8.08 × 109/L; Lymph% 40.1–67.8; Neutrophils# 1.59–6.41 × 109/L; Neut% 18.1–46.7; Basophils# 0.00–0.04 × 109/L; Baso% 0.0–0.4; red blood cell count 3.38–5.89 × 1012/L; haemoglobin (Hb) 116–198 g/L; Hematocrit % 35.2–61.2; mean corpuscular Hb (MCH) 30.9–36.3 pg/L; MCH concentration 310–341 g/L; mean corpuscular volume 94.5–112.2 fL; and platelets 210–610 × 109/L. The RIs had a conformity rate of 90%–100% in the validation specimens.Conclusion:We established region-specific RIs for CBC parameters in healthy newborns to help diagnose haematological disease in neonates.

Indexing hematological and serum biochemical reference intervals of Himalayan snow trout, Schizothorax esocinus to instrument in health assessment.

Schizothorax esocinus, commonly known as snow trout, is one of the main contributors of food and livelihood in the colder zone of Himalayan region. The comprehensive information on its hematological and serum biochemical reference intervals is not reported yet. In the present study an attempt has been made to elucidate the hematological and serum biochemical reference intervals of S. esocinus from River Jhelum using protocols of the American Society of Veterinary Clinical Pathology (ASVCP). Wild fish were sampled over a period of 2years from the pollution free sites of river Jhelum. Fish blood was harvested through caudal venipuncture and hemato-biochemical analysis performed thereof. Data values from a total of healthy 432 adult fish specimens (216 male, 216 female) were systematically recorded. The reference intervals for hematological and serum biochemical parameters of S. esocinus were established using Reference Value Advisor software v 2.1. RIs for hematological and serum analytes ranged as: hemoglobin (Hb) 78.38-116.35 (g/L); white blood cells (WBC) 10-20 (×109/L); red blood cells (RBC) 1.30-2.15 (×1012/L); packed cell volume 27.00-39.45 (%); total protein 39.21-61.62 (g/L); albumin 8.20-22.02 (g/L); globulin 27.58-49.55 (g/L); glucose 3.25-7.18 (mmol/L); urea 0.96-2.38 (mmol/L); cholesterol 3.80-6.90 (mmol/L). The study also depicted that certain blood measurands were influenced with respect to sex. Significantly (p < 0.05) higher values of Hb, red blood cells count and serum glucose were noted in male as compared to female which, on the other hand, registered higher white blood cells count and serum cholesterol level (Mann Whitney U test, p < 0.05). The work, therefore, provides baseline information on hematological and serum biochemical analytes of this species which holds high commercial importance. RIs reported here can help monitor the health status of fish by improving the use of non-lethal diagnostic methods in piscine medicine.

Hematology, Biochemistry, and Protein Electrophoresis Reference Intervals of Western European Hedgehog (Erinaceus europaeus) from a Rehabilitation Center in Northern Portugal

The Western European hedgehog (Erinaceus europaeus) can work as a bioindicator of environmental pollution and be a host for multiple zoonotic agents, making it relevant in terms of One Health studies. It is essential to deepen the knowledge on this species and calculate reference intervals (RIs) for the usual hematological and biochemical parameters. For this retrospective study (2017-2022), the archives of the Clinical Pathology Laboratory (LPC) of University of Trás-os-Montes and Alto Douro (UTAD) Veterinary Teaching Hospital were analyzed. Data of hematology, clinical biochemistry, and protein electrophoresis from 37 healthy hedgehogs of the Wild Animal Rehabilitation Center at UTAD, Northern Portugal, were included. It was possible to calculate RIs for almost all of the variables in the study, using Reference Value Advisor V2.1. Moreover, sex and age effects were investigated: alkaline phosphatase (p = 0.012, higher in males); total proteins (p = 0.034, higher in adults); mean cell volume (p = 0.007) and mean corpuscular hemoglobin (p = 0.010) (both higher in juveniles); and red blood cell distribution width (p = 0.021, higher in adults). Our study allowed for the first time to define RIs for a population of hedgehogs in Portugal, having a potentially relevant impact on species conservation and in the human-animal health interface.

Establishing pregnancy-specific haematological reference intervals in Ghana; a three-center cross-sectional study.

Abnormal intra-pregnancy haematological variables are associated with adverse feto-maternal outcomes. However, the reference intervals (RIs) employed in sub-Saharan Africa to inform clinical decisions are generally imported. Since RIs are influenced by age, geographical location, and race, we hypothesized that context specific RIs should be established in Ghana to contextualize intra-pregnancy decision making. This cross-sectional study retrospectively retrieved data of 333 pregnant women with no known clinically determined intra-pregnancy complications; 22 participants in their first trimester (T1; 1-13 weeks), 177 in their T2 (14-27 weeks), and 132 in T3 (28-41 weeks). RIs for haematological parameters were non-parametrically determined at 2.5th and 97.5th percentiles in accordance with CLSI guidance document EP28-A3c. Two-sample comparisons were undertaken using Wilcoxon rank-sum tests whereas more than two-sample comparisons were undertaken using Kruskal-Wallis test. Statistical significance was set at p <0.05 under the two-tailed assumptions. In accordance with WHO trimester-specific haemoglobin cutoffs, anaemia prevalence was a moderate (T1: 36.4%; 8/22 & T2: 31.6%; 56/177) to severe (T3:68.0%; 90/132) public health problem. Additionally, 9.3% (31/333) individuals had high gestational haemoglobin levels (Hb >13.0 g/dL). Moreover, haemoglobin (T2: 8.6-14.3 vs T3: 7.5-13.6 g/dL), MCH (T2: 22.5-69.8 vs T3: 21.6-31.9 pg), MCHC (T2: 30.2-51.8 g/L vs T3: 30.5-37.9 g/L), TWBC (T2: 4.0-13.4 vs T3: 4.1-13.0 x 109/L) required trimester specific RIs, compared to RBC (2.8-5.1 x 1012/L), MCV (66.2-100.2 fL), and platelet counts (106.3-388.3 x 109/L) that each required combined reference intervals. The intra-pregnancy haematological RIs determined have appreciable lower limits; there is the need to determine context-specific thresholds for haematological variables predictive of positive and/or adverse maternal and infant health outcomes.

Reference Intervals and Percentiles for Hematologic and Serum Biochemical Values in Captive Bred Rhesus (Macaca mulatta) and Cynomolgus Macaques (Macaca fascicularis).

Several physiological characteristics and housing conditions are known to affect hematologic and serum biochemical values in macaques. However, the studies that have been conducted either report values calculated based on a small number of animals, were designed specifically to document the effect of a particular condition on the normal range of hematologic and serum biochemical values, or used parametric assumptions to calculate hematologic and serum biochemical reference intervals. We conducted a retrospective longitudinal cohort study to estimate reference intervals for hematologic and serum biochemical values in clinically healthy macaques based on observed percentiles without parametric assumptions. Data were obtained as part of the Biomedical Primate Research Centre (Rijswijk, The Netherlands) health monitoring program between 2018 and 2021. In total, 4009 blood samples from 1475 macaques were analyzed with a maximum of one repeat per year per animal. Data were established by species, gender, age, weight-for-height indices, pregnancy, sedation protocol, and housing conditions. Most of the parameters profoundly affected just some hematologic and serum biochemical values. A significant glucose difference was observed between the ketamine and ketamine-medetomidine sedation protocols. The results emphasize the importance of establishing uniform experimental groups with validated animal husbandry and housing conditions to improve the reproducibility of the experiments.

Establishment of Complete Capillary Blood Counts Reference Intervals for Young Children in Local Area, China.

Blood count reference intervals are important to diagnose diseases and assess overall health, especially for young children. Although, in 2021, the National Health Commission of the People's Republic of China issued "Reference intervals of blood cell analysis for children (WS/T 779-2021)", these RIs may not suitable for small children all over the country due to racial, lifestyle, and geographical differences. The aim of this study was to establish and validate locally determined hematological reference intervals among young children in Nantong district and compare them with WS/T 779-2021 and American data. The reference sample consisted of 4,758 apparently healthy small children aged from age 28 days to 3 years according to the EP28-A3c guideline issued by the Clinical and Laboratory Standards Institute (CLSI). Capillary blood samples collected in K2-EDTA anticoagulant tubes analyzed by standard procedures. Statistical analysis was based on the guidelines of the CLSI. Pediatric reference intervals for 18 capillary complete blood count (CCBC) parameters were established for young children. WBC and differentials did not differ by gender in the combined analysis of all data, but showed some variations among different age groups, especially for NE and LYM. RIs of RBC value, MCV, and MCH were established, especially with regard to the difference among different age and gender groups. An overall increasing trend of PLT value was observed in children with no obvious difference between boys and girls. Further validation with 1,136 healthy subjects demonstrated that the verified proportions of our study were within 90.11% - 100%. RIs determined in the present study were more concentrated than WS/T 779-2021, with slight differences in the upper and bottom boundaries. Establishing appropriate region-specific reference intervals for pediatrics is essential. This study offers local reference intervals of CCBC values for young children and could be used as a benchmark for similar populations in the Yangtze River Delta economic region.

Determining Hematological Reference Intervals of the Blood Count in a Healthy Moroccan Adult Population (Al Hoceima Province).

The hemogram is the most frequently prescribed laboratory test. It guides the complementary tests essential to the diagnosis and follow-up of the patient. Hematology reference values can be influenced by several parameters such as environmental and genetic factors, rendering it essential to define reference intervals (RIs) for specific populations. This study aimed to determine RIs from a population of healthy adults in Al Hoceima province by following the procedures recommended by the IFCC-CLSI guidelines in 2008 and comparing them to those of similar studies conducted in various countries. We initially recruited 977 healthy adults from 18 to 60 years old including 255 men and 722 women, who presented themselves at the provincial hospital center of Al Hoceima (Morocco) for health checkups from February 2019 to September 2019. The erythrocyte, leukocyte, and platelet parameters were investigated using hematology analyzer ABX Pentra XL80 HORIBA® (HORIBA ABX SAS, Kyoto, Japan). The results showed that the RIs of the blood count parameters searched were more or less close to the RIs determined in Caucasian populations. Nevertheless, they were low compared to the limits of the RIs published in the literature for certain erythrocyte and platelet parameters. Our RIs were also similar in their majority, except for some parameters, to the RIs published very recently in a study conducted on a Moroccan population in the Tangier-Tetouan region. In addition, our intervals differ in their majority from those published in a study of a Ghanaian population. The differences reported by this preliminary work reinforce the need to establish hemogram RIs specific to the Moroccan population through more extensive studies carried out in different regions of Morocco. These studies should avoid misdiagnosis and allow physicians to interpret hematological tests more specifically.

Establishment of preliminary reference intervals and cytochemical staining of blood cells in big-bellied seahorses (Hippocampus abdominalis).

Despite their popularity, hematology reference intervals (RIs) have not been established in big-bellied seahorses (Hippocampus abdominalis). The objectives of this study were to establish hematologic RIs to compare values between sex in regard to cytochemical staining of blood cells. We also sought to compare white blood cell concentrations using the Natt and Herrick technique vs blood smear estimates. Forty-three healthy individuals from the Aquarium du Québec (22 females and 21 males) were included. Normal health status was confirmed by an unremarkable physical examination in five individuals and by necropsy of five other individuals, of which all were excluded from further analyses. Venipuncture was performed from the ventral coccygeal vein in the remaining 33 individuals without anesthesia using heparinized insulin syringes. A blood volume of 0.05 to 0.1ml was collected to prepare Wright Giemsa-stained blood smears and hematocrits immediately after venipuncture. Whole blood was stored in heparinized Eppendorf tubes to determine red and white blood cell concentrations using the Natt and Herrick technique with a hemocytometer in 10 individuals; these results were compared with blood smear estimates. Additional blood smears were stained with alkaline phosphatase substrate, periodic acid Schiff, and toluidine blue stains. The reference intervals included the packed cell volume (27.4-67.5%), thrombocyte count (19.5-197.7 × 109 /L), and white blood cell (WBC) count (2-54.8 × 109 /L), including neutrophils (1.1-21.3 × 109 /L), lymphocytes (2.7-45.5 × 109 /L), and monocytes (0-2.2 × 109 /L). The WBC hemocytometer counts showed no correlation with blood smear estimates (Spearman's rho=0.2). There was also no significant difference between the sexes. These preliminary reference intervals will help assess the health of seahorses.

Hematologic Reference Intervals and Comparison of Natt-Herrick Technique and Smear-Based Leukocyte Estimation in Cockatiels (Nymphicus hollandicus)

Although cockatiels are among the most common avian species maintained as companion animals in the United States, information on standard hematologic reference values for this species is limited. The objectives of this study were to establish hematologic reference intervals (RI) for cockatiels, compare methods using both the Natt-Herrick technique (NHT) and the smear-based estimation technique (SBT), explore age and sex differences in the hematologic findings for this species, and produce the first cockatiel RI for fibrinogen concentration and thrombocyte estimate. Healthy cockatiels (60 males and 60 females, 2-11 years old) from a research colony were included in this study. Blood samples were placed in dipotassium ethylenediaminetetraacetic acid tubes, and erythrocyte counts and thrombocyte estimates were determined via automated analyzer (ADVIA 120) and SBT, respectively. Moreover, leukocyte concentrations were determined using both NHT and SBT to compare these common methods for measuring a complete blood count in cockatiels. Data were analyzed for outliers, distributions, descriptive statistics, and RI via Reference Value Adviser, a set of macroinstructions for Microsoft Excel (Microsoft, Redmond, WA, USA). Lymphocytes were the predominant leukocyte across both methods. According to the NHT, females had significantly higher concentrations of total leukocytes, heterophils, bands, lymphocytes, basophils, and total plasma protein compared with males. Significant inverse polynomial relationships were noted between total leukocyte count and age and lymphocyte counts and age for NHT. Total leukocyte count produced via NHT and SBT were compared using Passing-Bablok and Bland-Altman plots, and no significant constant or proportional biases were found. However, these methods showed wide limits of agreement. While the RI were interchangeable between methods from a clinical standpoint, the same method should be used to assess changes in an individual. The reported RI are uniquely robust given the sample size, balanced sex and age distributions, inclusion criteria, and control over sample collection.