Abstract
THE following simple method modified from Jacoby (1944) was found suitable for the growth in vitro of Theileria annulata. A small fragment of infected calf's spleen or lymphatic gland is placed on a sterile glass coverslip and a drop each of calf plasma and chickens' embryonic extract are added. After coagulation the coverslip is sealed to the bottom of a large Carrel flask by a drop of plasma and embryonic extract; 3–5 c.c. of a mixture of 30–40 per cent calf serum in Tyrode is then added and the flask closed and incubated. On this medium Koch bodies survive for at least twelve days, but do not multiply. If fragments of normal spleen are placed in juxtaposition to the infected fragment at intervals of 3–5 days, Koch bodies survive for 15–18 days, but there is no obvious multiplication. The addition of glutamine (3 γ per c.c), pyridoxin (0·6 γ per c.c.), inositol (4 γ per c.c.) and riboflavine (0·4 γ per c.c.) to the mixture of serum and Tyrode induces multiplication of Koch bodies, which was observed in ten successive fragments of normal calf spleen during a period of two months. The addition of these factors to the transplant, in which Koch bodies have survived without multiplication for eighteen days, immediately induced multiplication of the surviving parasites.
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TCHERNOMORETZ, I. Multiplication in vitro of Koch Bodies of Theileria annulata. Nature 156, 391 (1945). https://doi.org/10.1038/156391a0
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DOI: https://doi.org/10.1038/156391a0
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