Blood chemistry reference values in racing pigeons

Avian Pathology

ISSN: 0307-9457 (Print) 1465-3338 (Online) Journal homepage: http://www.tandfonline.com/loi/cavp20

Blood chemistry reference values in racing pigeons (columba livia domestica) J.T. Lumeij & J.J. de Bruijne To cite this article: J.T. Lumeij & J.J. de Bruijne (1985) Blood chemistry reference values in racing pigeons (columba livia domestica) , Avian Pathology, 14:3, 401-408, DOI: 10.1080/03079458508436241 To link to this article: http://dx.doi.org/10.1080/03079458508436241

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Date: 28 January 2016, At: 14:33

Avian Pathology, 14: 401-408, 1985

BLOOD CHEMISTRY REFERENCE VALUES IN

RACING PIGEONS (COLUMBA LIVIA DOMESTICA) J.T. LUMEIJ and J.J. de BRUIJNE

Avian Pathology 1985.14:401-408.

Small Animal Clinic, State University of Utrecht, Yalelaan 8, 3584 CM Utrecht, The Netherlands SUMMARY Reference values for 23 blood chemical parameters in racing pigeons (Columba livia domestica) were established for use in clinical pathology. The inner limits are given for the percentiles P 2 . 5 and P 9 7 . 5 with a probability of 90%. A minimum of 50 blood samples collected between September and January from different animals aged between 6 months and 12 years (median 1.5 years) was used for each parameter. Reference values obtained in the present study are compared with values published previously. INTRODUCTION The diagnostic application of clinical pathology in human and veterinary medicine is a well-established additional procedure for the establishment of a diagnosis in certain disease conditions. Earlier demands for large blood sample volumes and limited veterinary involvement in pet bird disease were major obstacles to a parallel development in avian medicine. The introduction of micro methods in clinical laboratories has removed a major obstacle to the application of clinical pathology in pet bird medicine. The use of clinical pathology in any species, however, is dependent upon an extensive knowledge of reference values obtained from clearly defined normal populations. The present study was undertaken to establish reference values for some blood chemical parameters in racing pigeons (Columba livia domestica), and to compare these with values published previously (Erdös and Fontaine, 1977, 1978; Parker and George, 1976; Ramis and Planas, 1978; Wagner, 1978). MATERIALS AND METHODS Experimental animals Racing pigeons from a wide variety of breeds were used. About half the number of birds had been used for racing and had been housed in our pigeon loft (Small Animal Clinic, State University of Utrecht, The Netherlands) for at least 2 months Received 23 August 1984 Accepted 24 April 1985

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J.T. Lumeij and J J . de Bruijne

prior to the investigation. The animals had not been exposed to stressful situations during this period (e.g. racing). They were given about 30 g of a commercial pigeon ration (Witte Molen, Meeuwen, The Netherlands) each day at 13.30 hour (M.E.T.) and water ad libitum. The remainder of the birds belonged to the breeding stock of a commercial racing pigeon breeding farm (Eijerkamp B.V., Zutphen, The Netherlands).


On clinical examination all pigeons appeared to be healthy. Throat swabs were negative for trichomonads and faecal examinations were negative for parasite ova, oocysts and salmonellae. All pigeons were vaccinated against paramyxovirus disease with an inactivated Newcastle disease vaccine (Newcavac Nobilis, Mycofarm B.V., de But, The Netherlands) at least 1 month prior to the experiment. The age of the birds ranged between 6 months and 12 years (median 1.5 years) and their weights between 456 and 610 g (median 518 g). Blood sampling was done between September and January to exclude changes in blood chemical parameters in relation to the breeding cycle and at evenly spaced times between 08.00 and 18.00 (M.E.T.) to exclude confounding data due to possible variations arising from a circadian rhythm in the metabolic status of the birds or due to a postprandial effect. AU birds were exposed to the local day-night rhythm. A minimum of 50 blood samples from different animals was used to establish reference values for each parameter. Male and female pigeons were approximately equally represented. Sampling Samples were collected from the vena metatarsalis plantons superficialis medialis. To expose the vein the overlying skin was rubbed with a piece of cotton wool moistened with diethyl ether. The vein was punctured with a blood lancet and the blood was allowed to flow freely into blood collecting tubes. For serum total protein estimations and serum protein electrophoresis the blood samples were allowed to coagulate for 1 hour in plain blood collecting tubes before centrifuging, to prevent latent fibrin formation in the serum. For glucose estimations blood was collected in tubes containing 4 mg fluoride and 2 mg K2EDTA for 2 ml of blood. All other parameters were determined in plasma. Blood was collected in tubes containing 45 USP units of lithium heparin per 3 ml of blood. Immediately after drawing of blood plasma and cells were separated by centrifuging. Chemistry All analysis were done on the same day the samples were taken, except uric acid, iron and iron-binding capacity, for which samples were stored at -20°C. All enzyme determinations were carried out according to the Recommendations of the German Society of Clinical Chemistry (1972). The analytical methods and the coefficients of variation (V.C.) showing the day to day variation of these methods are tabulated (Table 1). Statistics Reference values were established using a distribution free method (Conoven, 1971; Riimke and Bezemer, 1972). The inner limits are given for the percentiles P 2 , 5 and P97.5 with a probability of 90%. To make comparison with data from other authors possible results are also expressed as mean ± s.cmean.

Table 1. Plasma or serum chemistry reference values for racing pigeons.


n

P2.S-P97.5 3

Median

Sodium Potassium Calcium Magnesium Anorganic phosphorus Chloride Plasma Iron

68 52 52 50 53 55 50

141-149 mmol/1 3.94.7 mmol/1 1.9-2.6 mmol/1 1.1-1.8 mmol/1 0.57-1.33 mmol/1 101-113 mmol/1 11-33 jUmol/1

145 4.4 2.2 1.3

Iron binding capacity

50

Osmolality Glucose Creatinin Uric acid CK (EC 2.7.3.2.) AP (EC 3.1.3.1.) LD (EC 1.1.1.27) ASAT (EC 2.6.1.1) ALAT(EC2.6.1.2) Total Protein (serum) Prealbumin Albumin a globulin ß globulin 7 globulin

55 96 52 50 50 65 50 50 50

Mean ± s.e.mean V.C.%h

Materials and methods

107 22

145 ±0.3 4.4 ±0.06 2.3+0.03 1.3 ±0.03 0.83 ±0.04 107 ±0.5 23 ±2.1

0.9 1.4 2.1 2.1 3.0 3.2 4.7

flame photometry flame photometry atomic absorption spectrophotometiy atomic absorption spectrophotometry colorimetry, TC 124 974 BMC coulometry, amperometry colorimetry, Guanidine-ferrozine method,

3045 pnol/1

37

37.5 ±0.8

5.2

colorimetry, Guanidine-ferrozine method,

297-317 m0sm/kg 12.9-20.5 mmol/1 23-36 jumol/1 150-765 jumol/1 130-380 U/l 160-780 U/l 32-107 U/l 40-85 U/l 18-31 U/l 21-33 g/1 14g/l 13-22 g/1 2-3 g/1 3-6 g/1 1-3 g/1

305

306 ±0.9 16.6 ±0.2 28 ±0.6 375 ±30 203 ±13.5 367 ± 25 57 ±3.2 58.6 ±2.5 25+1.6 27 ±0.4 2.8 ±0.13 17.5 ±0.36 2.29 ±0.08 4.3 ±0.14 1.9 ±0.1

4.7 3.2 2.4 4.0 4.6 4.3 6.0

vapour pressure osmometry MCAd, Hexokinase method, TC 124 338 BM MCA, Jaffékinetic reaction at 30° C. photometry, uricase method TC 124 737 BM MCA,30°C,TC181 188 BM MCA,30°C,TC415 278BM MCA, 30° C, TC 158 186 BM MCA,30°C,TC158 178BM MCA, 30° C, TC 244 236 BM MCA, biuret method with blank correction

0.81

TiaVpr fTipm

Tialcer Phem

a c

93 58 58 58 58 58

The inner limits for P2.5 and P97.5 are given. Test Combination Boehringer Mannheim, West Germany.

16.4 27 330 170

320 50 55 23

28 2 17 2 4

2

5.1 3.8

2.9

I o

•o

I Serum protein electrophoresis was performed using cellulose acetate membranes. Barbital > buffer pH 8.6, 20 minutes, 200 Volts. Staining with Ponceau S

J

Day to day variation coefficient of analytical method. Multistat FLS Centrifugal Analyser. Allied Instrumentation Laboratory, London, U K.

4

8

404

J.T. Lumeij and J J . de Bruijne

RESULTS Reference values for 23 plasma or serum chemical parameters of racing pigeons, as established in this study, are shown in Table 1. Fig. 1 shows a densitometer scan and the electrophoretic pattern from a representation pigeon serum. Plasma glucose concentrations in morning samples were significantly lower than plasma glucose concentrations in samples taken between 14.00 and 18.00 hr


DISCUSSION A subdivision in age-groups, sex and time of sampling would have been impractical for clinical use, although variation due to these factors cannot be excluded. The wide belief that biological data assume a Gaussian distribution is inappropriate (International Federation of Clinical Chemistry, 1978). Also in this study most of the data could not be described by a Gaussian distribution. Therefore non-parametric statistics had to be used to establish reference values (International Federation of Clinical Chemistry, 1984). However, a less appropriate parametric estimation technique was also applied (mean ± s.e.mean) to enable comparison with reference values from other authors who did not give information about distribution of data and may have misused this method in ignorance of its basic assumptions. The plasma uric acid concentration as determined in this study was 375 ± 30 jumol/1 (mean ± s.e.mean). Wagner (1978) found plasma uric acid concentrations of 219 ± 13 jjmol/1 (mean ± s.ejnean) in white Carneaux pigeons. These lower values might be attributed to the fact that this parameter was measured in fasting subjects. Erdös and Fontaine (1977) determined reference values for blood chemical parameters in healthy racing pigeons but did not include uric acid in their study. In analogy with mammals they determined serum urea concentrations as a parameter for renal function. Because pigeons are uricotelic it seems more appropriate to measure plasma or serum uric acid concentrations to assess renal function in this species. The plasma potassium concentrations in the present study was 4.4 ± 0.06 mmol/1 (mean ± s.ejnean). Erdös and Fontaine (1977) have found serum potassium values of 3.8 ± 0.1 mmol/1 (mean ± s.e.mean, n = 20) in 4-year-old female racing pigeons. However, in their study the serum was separated from the cells after the blood samples had been standing for 1 hour at room temperature. In a recent study (Lumeij, 1985) it has been shown that plasma potassium values in pigeons start to decrease immediately after collection of the blood sample. It was demonstrated that reliable results could only be acquired if plasma and cells were separated immediately after collection. Erdös and Fontaine (1977), using the biuret reaction for the determination of serum total protein, give reference values for racing pigeons within the range of the reference values presented in this article. The electrophoretic results, however, cannot be compared because different materials and methods were used. In this study a plasma glucose concentration of 16.6 ±0.2mmol/1 (mean±s.e.mean) was found. Morning values were lower than afternoon values, suggesting a circadian

405


Blood chemistry of racing pigeons

Pre-alb. Alb.

a ß

y

Fig. 1. Densitometer scan and electrophoretic pattern from a representative pigeon serum. Pre-albumin, albumin, a-, ß-and y globulin.

Blood chemistiy of lacing pigeons

407

rhythm in plasma concentrations or a postprandial hyperglycaemia in this species. The reason that other authors report lower reference values for plasma glucose values in racing pigeons might be associated with different times of sampling. Erdös and Fontaine (1978), for example, report values of 10.8 ± 0.4 mmol/1 (mean ± s.e.mean, n = 20) for plasma glucose in 4-year-old female racing pigeons. Sampling was done between 8 a.m. and 9 ajn. (Köln/Germany, February). According to our findings, values in the lower range are to be expected at that time of the day. Parker and George (1976) reported a plasma glucose concentration of 10.1 ± 0.15 (mean ± s.ejnean, n = 26) in randomly bred white Carneaux pigeons {Columba livid). The lower values reported by the latter authors might be associatewith breed differences. The question whether a circadian rhythm can be found in plasma glucose concentrations and the existence of a postprandial hyperglycaemia in pigeons should be investigated.


Reference values for plasma iron were in agreement with values published previously by Erdös and Fontaine (1978) and Ramis and Planas (1978). Reference values for total iron-binding capacity however, were markedly lower than reported by the above mentioned authors. No explanation was found. Reference values for ALAT, ASAT and AP cannot be compared with values reported previously (Erdös and Fontaine, 1978) because different methods were used (Bessey, Karmen and Wrobléwski respectively). These methods can be considered less accurate and inappropriate to modern instrumentation.

REFERENCES Erdös, A. und R. Fontaine (1977). Ermittlung von Notmalwerten des Blutes bei drei Taubenrassen in Abhängigkeit von Alter und Geschlecht. 1. Mitteilung. Archiv für Geflügelkunde 41, 238-245. Erdös, A. und R. Fontaine (1978). Ermittlung von Normalwelten des Blutes bei drei Taubenrassen in Abhängigkeit von Alter und Geschlecht. 2. Mitteilung. Archiv für Geflügelkunde 42, 34-44. Conoven, WJ. (1971). Practical Nonparametric Statistics. John Wiley & Sons, New York. International Federation of Clinical Chemistry (1978). The concept of reference values. Clinica Chimica Acta 87, 459F-465F. International Federation of Clinical Chemistry (1984). Statistical treatment of collected reference values. Determination of reference limits. Clinica Chimica Acta 137, 97F-114F. Lumeif, J.T. (1985). The influence of blood sample treatment on plasma potassium concentrations in avian blood. Avian Pathology 14, 257-260. Parker, GA. and J.C. George (1976). Changes in blood glucose, blood and muscle lactate, and plasma lactic dehydrogenase levels in the pigeon on acute exposure to cold. Archives Internationales de Physiologie et de Biochimie 84, 517-526. Ramis, J. and J. Planas (1978). Iron metabolism in pigeons. Quarterly Journal of Experimental Physiology 63, 383-393. Recommendations of the German Society for Clinical Chemistry (1972). Journal of Clinical Chemistry and Clinical Biochemistry 10, 281-291. Rilmke, C.L. and P.D. Bezemer (1972). Methods for the determination of normal values II New methods. Nederlands Tijdschrift voor Geneeskunde 116, 1559-1568. Wagner, W.D. (1978). Risk factors in pigeons genetically selected for increased atherosclerosis susceptibility. Atherosclerosis 31, 453-463.

RESUME Valeurs de référence des constantes biochimiques du sang chez le pigeon voyageur (Columba livia domestica) Des valeurs de référence concernant 23 paramètres biochimiques du sang chez

408

J.T. Lumeij and J J. de Bruijne

les pigeons voyageurs (Columba livia domestica) ont été établies pour être utilisées en pathologie clinique. Les limites intérieures sont données pour les percentiles P 2 : 5 et P9 7 . 5 avec une probabilité de 90%. Un minimum de 50 échantillons prélevés entre septembre et janvier à partir des différents animaux âgés de 6 mois à 12 ans (medianne 1.5) ont été utilisés pour chaque paramètre. Les valeurs de référence obtenues dans la présente étude sont comparées avec les valeurs publiées précédemment.


ZUSAMMENFASSUNG Referenzwerte für die Blutchemie bei Reisetauben (Columba livia domestica) Für den Gebrauch in der klinischen Pathologie wurden Referenzwerte von 23 Blutparametern bei Reisetauben (Columba livia domestica) aufgestellt. Die Gültigkeit wurde beschränkt auf die Prozentwerte zwischen P 2 . 5 und P 9 7 . 5 mit einer Wahrscheinlichkeit von 90%. Ein Minimum von 50 Blutproben wurde zwischen September und Januar von verschiedenen Tieren im Alter zwischen 6 Monaten und 12 Jahren (Mittelwert 1.5 Jahre) für die Bestimmung jeden Parameters ausgewertet. Die in dieser Untersuchung gewonnenen Referenzwerte wurden mit anderweitig veröffentlichen verglichen. RESUMEN Valores de referencia de la quimica sanguinea en palomas de Carreras Los valores de referencia para 23 parámetros de química sanguínea en palomas de carrera (Columba livia domestica) fueron establecidos para su uso en patología clínica. Los límites internos son expresados por los porcentajes P 2 . 5 y P 9 7 . 5 con una probabilidad del 90%. Un m'inimo de 50 muestras de sangre colectadas entre septiembre y enero a partir de animales diferentes, con edades de 6 meses a 12 años (promedio de 1.5 años) fueron empleados para cada parámetro. Los valores de referencia obtenidos en el presente estudio son comparados con los valores previamente publicados.