Camp. Biochm. Physiol. Vol. 98A. No. 3/4, pi. 393-394, 1991 Printed in Great Britain

a~~~~1 53.00 + 0.00 Q 1991 Pergamon Press pk

HEMATOLOGY AND BLOOD CHEMISTRY OF THE WHITE PELICAN (P~L~CA~US O~~C~OTAL~~)

M. L. PUERTA, A. L. GARCIA DEL CAMPO,+ V. HUECM and M. ABELENDA

Departamento de Biologia Animal II (Fisiologia Animal), Facultad de Ciencias Biologicas, Universidad Complutense, 28040 Madrid, Spain

(Recebvd 27 June 1990)

Abstract-l. The hematology and blood chemistry of six captive White Pelicans (Pe1ecnnu.t ancpcroralus) was studied.

2. The number of erythrocytes was 2,5OO,OOO/mm’, the hematocrit was 41.5% and the blood hemoglobin content was 13.9 g/l00 ml.

3. Leukocyte number was 13,500~mm3, the differential counts being heterophik 59.5%, eosinophiis 3.1%, basophils 1 .O%, lyrnph~yt~ 35.3% and monocytes 1.1%. ~rom~yte number was 4~~mm3.

4. Plasma composition was (mg/lOO ml): glucose: 191; triglycerides: 61; cholesterol: 99; phospholipids: 166; urea: 7.8; uric acid: 5.5; creature: 0.5 and bilirubin: 0.1.

5. Total protein concentration was 3.4 g/100 ml. 6. Enzymatic activities (U/l) were: GOT: 268; GPT: 26; LDH: 994; CK: 583; y-GT: 9-19 and LAP: 57.

I~RODU~ON

Pelicans are almost cosmopolitan. Migratory or dis- persive, there is little modem information on routes or winter quarters of either European breeding species. The White Pelican (Pelecanus onacrotaius) is one of the largest Pelicans whose presence has never been recorded in Spain (Cramp and Simmons, 1977). As far as we know, there is no data about its physiology. This paper deals with its hematology and blood composition.

MATERIALS AND METHOD!3

This study was carried out with six adult Pelecanus onocrotalus which were captive in the Madrid Zoo. At the time of blood sampling (October, 1989) they had spent 3 years in the zoo. They could move around the zoo but were unable to fly as a consequence of a surgical intervention. Animals were lightly anaestheaized by inhalating isofluorane and a blood sample (0.5-I ml) was collected from the radial vein in heparimzed syringes (25 U/ml). Aliquots of blood were diluted (ZOO and 50 times for red and white cells, respectively) in hematological pipettes with Natt and Her- rick’s (1952) solution. The number of erythrccytes and leukocytes was counted in 96 small squares and all the large squares of cell counting Thoma chambers, respectively. Hematocrit was measured by means of capillary tubes. Hemoglobin was assayed according to the calorimetric method of Drabkin (1945). Blood smears were fixed (3 min immersion in methanol) at the time of blood coflection. They were stained with commercial Giemsa stain (Merck, West Germany) diluted 1:4.5 (v/v) in phosphate buffer, pH 6.8, for 45 min. Identification and counting of leuko- cytes and thrombocytes were done with a tight microscope with oil immersion lens ( x 100). At least 200 white cells were counted in each sampb. The number of ~rorn~yt~ was transformed into absolute numbers by reference to the total number of white blood cells. No attempt was made to count

*Veterinary of the Madrid Zoo, Casa de Campo, 28011 Madrid, Spain.

immature forms of erythrocytes, or to different~te large and small lymphocytes. Plasma was obtained by centrifugation and stored at -20°C until analysis. All the biochemistry tests were performed in duplicate, using an autoanalyser Hitachi 737. Values in the text are mean + SE.

RESULTS AND DISCUSSION

Heterophils and cosinophils showed a trilobulatcd nucleus. Eosinophils showed densely packed or- ange-pink granules which were not distinguished individually. Heterophils showed a uniform cyto- plasmic distribution of pale pink granules with the presence of what appeared to he a second type of granule, these latter being more scarce, rounded and deep pink in colour. Basophils granules and cyto- plasm stained deep blue.

Blood hemoglobin content, hematocrit, red blood cells (RBC) number and indices of the White Pelican are depicted in Table 1. They are coincident with those previously found in Brown Pelicans (Pefecanw occidentalis) that were captive in a zoo also located outside their natural geographic distribution (Balasch et al., 1974). The fact that two different species (P, onocrotalus and P. occidentah) with different natural distributions have similar values for RBC related parameters raises the possibility of a narrow range for these parameters in other Pelecams sp. It should be kept in mind that both the White Pelicans of the present study and the Brown Pelicans of the study of Balasch e? al. (1974) were captive and unable to fly. Captivity changes the blood composition (de- Graw et al., 1979) and probably also reduces oxygen demands for flying (Hunter and Powers, 1980; Gesaa- man et af., 1986). Therefore, another plausible hy- pothesis raised from this study could be that RBC parameters have been reduced to satisfy the oxygen demands of unflying animals and that these demands are similar in captive Pelicans of different species.

393

394 M. L. mItTa er al.

Tabk I. Hematology of captive Pelecanus onocroraltu 1980). On the other hand, although birds are said to Erythrocytes (cell/mm’) 2,546,OOO jz 83,000 Hematocrit (%) 41.5 f 1.0 Hemoglobin (g/l00 ml) 13.9 f I.1 MCHb (pg/cell) 54.9 f 4.5 MCHhC (g/l00 ml) 33.5 f 2.0 MCV (pm’) 163+6 Leuko&tes (cell/mm’)

HeteroDhils (%) Eosinobhils i%j Ftasophils (%) Lymphoctya (%) Monocytes (%)

13,500* 1100 59.5 f 10.2

be uticotelic, urea and uric acid are present in ap- proximately similar proportions in the plasma. This proportion differs from that found in captive Chilean Flamingos, which almost lack of urea in plasma (Puerta et al., 1989a) but it has also been recorded in other captive birds (unpublished results).

3.1 ; I.1 1.0 + 0.4

35.3 -+ 9.1 I.1 f 0.2

Acknowledgement-We gratefully appreciate the co- operation of Madrid Zoo.

Thrombocytes (cdl/mm’) 4500 + 1400

MCHb: mean all hemoglobin, MCHbC: mean cell hemoglobin concentration, MCV: mean cell volume. Thrombocytes were only counted in four animals.

Table 2. Plasma concentration of several blood comwnents in captive Pelecanu onocrotolu

Total proteins (g/100 ml) 3.4 + 0.3 Glucose (mg/l@I ml) 191 f 20 Triglycerides (mg/lOO ml) 61 27 Cholesterol (mg/lOO ml) 99+ IO Phospholipids (mg/lOO ml) 166kfl Urea (m&100 ml) 7.8 + 2.0 Uric acid (mg/lC%l ml) 5.5 2 0.5 Creatinc (mg/lOO ml) 0.5 f 0.0 Bilirubin (mg/lOO ml) 0.1 ?I 0.0 GOT (U/l) 268 + 52 GPT (U/I) 26 2 2 LDH (U/I) 994 k 258 CK (U/l) 583 f 76 y-GT (U/l) range t&19 LAP (U/I) 572 I2

GOT: glutamyl-oxala~tic traosaminase; GPT: glyamyl-pyruvate transaminase; LDH: lactic dc- hydrogenax; CK: creatine kinase; y-GT: gamma-glutamyl transferase; LAP: leucin amino peptidase.

To our knowledge, no previous study has dealt with the white blood cells (WBC) total and differen- tial counts of Pelicans. The White Pelican has 13,500 WBC/mm’ (Table l), heterophils (60%) being the most abundant WBC type followed by lympho- cytes (35%). This relative proportion differs from that usually assumed for birds (Hodges, 1979; Jones and Johansen, 1972; Sturkie, 1986) and has also been found in other birds both captive (Hawkey er al., 1983; Sturkie, 1986) and free-living (Puerta et al., 1990). Eosinophils, basophils and monocytes are in smaller proportions (Table 1). Thrombocytes number was small when compared with other birds (Sturkie, 1986; Puerta ef al., 1989b).

Plasma composition is depicted in Table 2. Total proteins are in the range of other captive birds but are greater than in the Brown Pelican. On the contrary, plasma glucose level is somehow smaller than in other captive birds but is similar to that of the Brown Pelican (Balasch ef al., 1974; Gee et al., 1981). Enzymatic activities do not differ markedly from those of other captive birds (Gee et al., 1981; For- shaw and Cooper, 1988; Puerta et al., 1989a). Triglyc- erides, cholesterol and phospholipids are smaller than in other birds both captive and free-living (Gee et al., 1981; Puerta et al., 1989a, 1990). Bilirubin and cre- atine are in the range of other captive birds (Rafael,

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