moditicd Wrighl-Giemsa slain, and 

 examined by oil immersion microscopy. 

 The red cells were round, moderate- 

 ly anisocytotic ranging from 7.5-9.5 /i 

 in diameter, and appeared well hemo- 

 globinated with only occasional slight 

 central pallor. An occasional red cell 

 displayed polychromasia. and some 

 rare Howell-Jolly bodies were seen. 

 No nucleated red cells were encoun- 

 tered. 



A white cell differential count was 

 as follows; 



Segmented neutrophils b^ percent 

 Band forms 19 percent 



Metamyelocytes < 1 percent 



Monocytes 9 percent 



Lymphocytes 8 percent 



No eosinophils nor basophils were en- 

 countered. The lymphocytes were all 

 of the large type. No small lympho- 

 cytes with the typically scant cyto- 

 plasm and dark staining nuclei were 

 present. Twenty-one percent of the 

 nuclei of the mature, segmented neu- 

 trophils had distinct '"drumstick" ap- 

 pendages. 



The thrombocytes appeared as 

 round platelets, with diameters ap- 

 proximately one-third to one-half of 

 those of the red cells. Their number, 

 estimated from their frequency distri- 

 bution on the smear in relation to the 

 erythrocytes, was in the range of 

 300.000-350,000/cubic mm. 



HEMOGLOBIN 

 ELECTROPHORESIS 



Hemoglobin electrophoresis was 

 performed by the vertical acrylamide 

 gel technique as described in detail 

 elsewhere (Bierman and Zettner. 1967) 

 (Nakaniichi and Raymond. 1963). 

 Briefly, a toluene hemolysate of the 

 washed red cells is prepared and elec- 

 trophoresed in Tris-buffer of pH 9,0 

 for 3'/2 hours at 120 ma. The acryla- 

 mide gel slabs are then stained with 

 amido black and destained electro- 

 phoretically in 5 percent acetic acid. 



The results are shown in Figure 1. 

 The whale hemoglobin (Slots Nos. I 

 and 7) migrated slightly slower than 

 human hemoglobin A. The position 



SLOT I 



3 4 5 6 7 



6-9'71 



Whale non-heme protein 

 (Whale Carbonic Anhydrase ?) 



Human Corbonic Anhydrase B 



Humon HbAg 

 Human HbS 



Gray Whale Hb 

 Human HbA 



fill • 



Figure 1. — Vertical acrylamide gel hemoglobin electrophoresis. Tris-buffer, pH 9.0. The gel contains 

 the toluene hemolysates of the following: Slots No. 1-gray whale; No. 2-human with A-S trait; No. 3, 4- 

 normal humans; No. 5.6-standards; No. 7-gray whale (same as slot No. 1). Original (sample 

 application slots) at top. Cathode-lop; anode-bottom. 



of the band of the whale hemoglobin 

 was indistinguishable from that where 

 human hemoglobin F-' would be ex- 

 pected. No minor hemoglobin com- 

 ponents equivalent to those found in 

 human blood could be detected. The 

 weakly stained band of much slower 

 mobility, as shown in Figure I, is a 

 non-heme protein, as indicated by the 

 failure of this protein band to react 

 with benzidine when a freshly electro- 

 phoresed, unstained strip of the gel 

 containing the whale sample was sub- 

 mersed in a benzidine and peroxide 

 solution. 



The pattern of hemoglobin electro- 

 phoresis performed on blood samples 

 obtained on 17 March and 27 April 

 1971, and 13 March 1972 was iden- 

 tical to that demonstrated here. 



ALKALI DENATURATION 



A quantitative alkali denaturation 

 test performed on the toluene hemoly- 

 sate by the method of Singer, Chernoff, 

 and Singer ( 195 1) revealed the whale's 

 hemoglobin to be 100 percent alkali 

 resistant. The alkali resistance of the 

 hemoglobin was the same in all sam- 

 ples obtained over the course of 1 

 year, as listed above. 



DISCUSSION 



The values of the various red cell 

 parameters, as reported here, are in 

 fair agreement with those published by 

 Lenfant (1969). Relative to most ter- 

 restrial mammals, the California gray 

 whale appears to have lower red cell 

 counts, hemoglobin concentrations, 

 and hematocrits, although the MCV 

 is considerably in excess of 100 jU^. 

 A proportional increase of the MCV 

 of red cells with total body length of 

 marine mammals of different species 

 has been shown (Lenfant. 1969). Of 

 interest is the finding of Lenfant ( 1969) 

 of a high proportion of nucleated red 

 cells in the gray whale. This is in 

 distinct contrast to the complete ab- 

 sence of nucleated red cells in the 

 blood samples examined here. It 

 should be considered that the previous 

 observations were apparently made on 

 sick, wounded, dying, or dead animals: 

 and that under these abnormal condi- 

 tions, immature red cells may have 

 been released into the circulation. 

 The only indication of young red cells 

 in our samples were the rare Howell- 

 Jolly bodies and occasional polychro- 

 masia. 



23 



