Plasma chemistry reference values in the gyr falcon (Falco rusticolus)
Morena Bernadette Wernick, Olga Martin-Jurado, Hugues Beaufrère & Jaime Samour Comparative Clinical Pathology ISSN 1618-5641 Comp Clin Pathol DOI 10.1007/s00580-013-1794-5
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Author's personal copy Comp Clin Pathol DOI 10.1007/s00580-013-1794-5
ORIGINAL ARTICLE
Plasma chemistry reference values in the gyr falcon (Falco rusticolus ) Morena Bernadette Wernick & Olga Martin-Jurado & Hugues Beaufrère & Jaime Samour
Received: 30 October 2012 / Accepted: 31 July 2013 # Springer-Verlag London 2013
Abstract Blood samples were collected from 102 clinically healthy gyr falcons (Falco rusticolus) over a time period of approximately 3 years as part of routine examination procedures. Standard blood chemistry analyses were carried out to establish normal reference values for the species. Plasma chemistry analyses included alkaline phosphatase (ALP), amylase, blood urea nitrogen, bile acids, calcium, cholesterol, creatinine, creatine kinase (CK), aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), glucose, phosphate, iron, total protein, total bilirubin, and uric acid. Reference intervals were determined using a quantile approach with 90 % confidence intervals of the limits. Juveniles and adults differ in ALT, creatinine, AST, glucose, LDH, and uric acid. Sexes differ in ALP, CK, AST, and phosphorus. This study provides valuable information on the plasma biochemistry values for gyr falcons that may help in the medical management of this endangered and commonly used falconry species. Keywords Gyr falcons . Falco rusticolus . Plasma chemistry . Reference values
M. B. Wernick (*) Tierklinik Thun-Süd AG, Burgerstrasse 11, 3600 Thun, Switzerland e-mail:
[email protected] O. Martin-Jurado AO Foundation, Clavadelerstrasse 8, 7270 Davos, Switzerland H. Beaufrère Health Sciences Centre, Ontario Veterinary College, University of Guelph, 50 stone road, Guelph, ON N1G2W1, Canada J. Samour Wrsan Wildlife Division, PO Box 77338, Abu Dhabi, United Arab Emirates
Introduction The gyrfalcon (Falco rusticolus) is the largest falcon, and its range spans the Arctic coast and islands of North America, Greenland, Europe, and Asia, inhabiting tundra and mountains, but also spends a considerable time on sea ice far from land during winter time (Burnham and Newton 2011). On average, males weigh 960–1,304 g, whereas females are larger and heavier, weighing 1,396–2,000 g (Dunning 2008). During the early 1970s, gyr falcons have been favored for Arab falconry because of their size and strength. As they were not able to acclimate with the extreme weather conditions prevalent in the Middle East, their popularity first decreased but came back as falcon traders of the emerging republics of the former Soviet Union tried to open new markets with Middle Eastern countries (Wernery et al. 2004). Even though a large number of gyr falcons are held in captivity, there is still a lack of biomedical and clinical pathology information on this species. Plasma chemistry analysis plays an important role in routine monitoring, assessment, and management of disease in avian patients. Reference values of clinically healthy birds are therefore necessary to evaluate biochemistry changes in specific pathological conditions. To the knowledge of the authors, plasma chemistry reference values for gyr falcons have been published in former studies (Altman et al. 1997; Lierz 2003), but several reference values are still lacking. Validity of results in former studies often remains questionable due to small sample sizes. According to the American Society for Veterinary Clinical Pathology (ASVCP), Quality Assurance and Laboratory Standards Committee (QALS), and the Clinical Laboratory Standards Institute (CLSI) guidelines for the determination of reference intervals in veterinary species, ideally a minimum of 120 reference individuals should be available for determining nonparametric RI and 90 % confidence intervals (CI) of reference limits (Horowiz et al. 2008, ASVCP 2011). Besides, plasma chemistry values are
Author's personal copy Comp Clin Pathol
sometimes reported in former studies without mentioning how they were obtained or values have been obtained with methods which are no longer considered as reliable in birds (Lumeij 2008). This paper describes the results of blood chemistry analyses, carried out on 102 clinically healthy gyr falcons at the Wrsan Wildlife Division, Abu Dhabi, United Arab Emirates.
Material and methods Blood samples were obtained from 102 apparently healthy gyr falcons during routine examinations over a time period of approximately 3 years at the Wrsan Wildlife Division in Abu Dhabi. The mean body weight of the birds was 970±17.43 g for the males and 1,384±23.52 g for the females (mean±SD), and their ages were between 4 months and 3 years. Thirtyeight males and 64 females were included in the study. All birds did not receive any medication for a minimum of 2 weeks before blood sampling and were used to manual restraint. The falcons were manually restrained and anesthetized with a mixture of 5 % isoflurane (Matrx, Spartan, V.M.C.) in 2 l of oxygen/min for the sampling procedure. An average of 2.5 ml of blood was obtained from the right basilic vein (vena cutanea ulnaris superficialis), using 3 ml disposable syringes fitted with 23 gauge×1 in. disposable needles. After collection, 2 ml of the whole blood was immediately mixed with the anticoagulant lithium heparin (1.8 mg/ml of blood) in commercially available storage tubes for blood chemistry analysis (BD, Franklin Lakes, NJ, USA). The birds were fasted over a time period of 12 h prior to blood collection. All samples were analyzed within 30 min after blood collection. No signs for hemolysis could be detected in any of the blood samples. A wet chemistry analyzer, Miura 500 (I.S.E. S.r.L., Roma, Italy), was used for blood chemistry analysis. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), creatine kinase (CK), gamma-glutamyl transferase (GGT), bile acids, total bilirubin, amylase, blood urea nitrogen, creatinine, uric acid, glucose, cholesterol, phosphate, iron, and calcium were determined with this device. The following techniques were used by the biochemistry device to determine the different biochemical values: enzymatic method for AST and ALT, enzymatic/colorimetric method for ALP, enzymatic/catalytic pyrvic method for LDH, kinetic reaction for CK, catalytic method for GGT, Diazyme enzymatic method for bile acids, coupling/hydrolysis of enzymes for amylase, urease method for BUN, Jaffe reaction for creatinine, uricase method for uric acid, glucose oxidase for glucose, hydrolysis into free cholesterol for cholesterol, colorimetric method for phosphorus (NH4 molybdate), oxidation for iron, and Arzenaso III method for the determination of calcium. According to the manufacturer, a
human-based calibration serum was used as a protein standard. Total plasma protein was measured by using the biuret method. Albumin was analyzed by using the bromocresol green dye. Globulin values and albumin/globulin ratios were assessed from total protein and albumin values. Statistical analysis All biochemical parameters were assessed for normality using a Shapiro–Wilk test and quantile plots and for homogeneity of variances across ages and gender using Levene test. Reference intervals were determined according to the CLSI guidelines, also endorsed by the American Society of Veterinary Clinical Pathology (Horowiz et al. 2008, ASVCP 2011). Ninety-five percent reference limits were determined as the interval between the 2.5 and 97.5 % quantiles. Ninety percent confidence intervals of the reference limits were obtained using a bootstrap approach to assess precision of these reference interval limits. Outliers were detected using Tukey method. The influence of age and sex was assessed using two-way ANOVA and Wilcoxon signedrank test when ANOVA assumptions were not met. For parameters that varied with age and gender, stratified reference intervals were not reported because the sample size would be too small for gender and age groups taken individually. R (R development core team (2012). R foundation for statistical computing, Vienna, Austria. http://www.R-project.org/) was used for statistical analysis and Reference Value Advisor for reference intervals determination (Geffré et al. 2011).
Results The results of the plasma chemistry study carried out on clinically healthy gyr falcons are presented in Tables 1, 2, and 3. There were significant differences between juveniles and adults for ALT (median, 69.71 and 33.92 for juveniles and adults, respectively, Wilcoxon rank-sum test, p
