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During the course of the routine laboratory examinations of leukemic cattle at the Veterinary Hospital Clinic of the University, Dr. M. Sonoda (personal communication) has chanced to see the presence of a considerable number of dividing cells in smears of isolated peripheral leucocytes from apparently normal animals. This prompted us to study the diagnostic feasibility of those mitotic cells to other mammalian species as well as to some pathologic conditions such as f reemartins, leukemias and congenital anomalies. After several preliminary trials we established a simple and useful technique to obtain good metaphase figures available for karyotype analysis in direct preparations from peripheral blood in several species of mammals, procedures of which will be described in this short communication. Peripheral blood was obtained from the carotid or other veins, or directly from the heart in 11 species of mammals listed in Table 1. For larger animals, the cow to the rabbit, about 3 ml of whole blood was mixed with 6-9 ml of Eagle's culture medium (MEM) containing 15 per cent bovine serum, while for smaller animals such as rats, mice and hamsters, 0.25-3 ml of whole blood was obtained. They were incubated for 2.5-5 hours at 37°C, and colchicine was added at concentrations of 0.2pg/ml (Indian munt jac) and 0.03 pg / ml (others) for final 2-3 hours. Chromosome slides were made according to the usual air-drying method after hypotonic treatment with 0.075 M KCl for 20 minutes at 37°C. By means of the hypotonic treatment and repeated changes of the fixative (3:1 methanol-acetic acid), red cells were completely broken and removed. The slides were stained with Giemsa. Usually 3 to 5 slides were prepared from one sample. Table 1 summarizes the results of observations. Mitotic index in each case was scored on the basis of 1,100-4,600 cells having a larger round-shaped nucleus, excluding mature granulocytes and small lymphocytes. Within the scope of the present study larger animals belonging to ungulates yielded good results, in which karyotype analyses were practically possible (Fig. 1, A to D). On the other hand, rodents, carnivores and

A SIMPLE METHOD FOR THE DIRECT CHROMOSOME DEMONSTRATION FROM PERIPHERAL BLOOD IN SOME MAMMALS