The nutritional requirement of Campylobacter fetus was investigated by static cultivation at pH 6.80 in the presence of 0.05% thioglycollate, which was added aseptically to avoid reduction of the growth-promoting effect due to heating. Remarkable multiplication was recognized by the coexistence of L-cystine and thioglycollate in a vitamin-free amino acid semisolid medium. L-Cystine was very important for growth, and the composition of a minimum number of amino acids was also required for growth, such as L-aspartic acid, L-glutamic acid, L-cystine and DL-methionine. Guanine among the nucleic acid bases brought about a distinctively heavy growth. Ribonucleic acid, ribose, glutathione, pimelic acid and a certain commercial brand of glycogen clearly showed growth-promoting effects as demonstrated by the increase in additional amounts in a conditional amino acid medium. Similarly, taurocholate acted as an effective growth promoter out of four bile salts tested. Guanine and ribonucleic acid were detected by paper chromatographyfrom the ingredients of media in common use for cultivation of C. fetus. A homemade yeast extract medium retained the typical cell form in almost all of the cultures during long term preservation at room temperature, in contrast to the production of various involution forms in commercial media and a meat extract medium. In conclusion, supplies of both thiol and disulfide groups fulfilled an important role by bringing into action some additives as nutritive substances to be utilized by C. fetus.For the past 10 years, investigations on the cultivation of Campylobacter fetus have been carried out by many workers. Above all, the noteworthy efforts was the elucidation of the optimal microaerophilic gaseous environment for multiplication [10,15,29]. In this study large quantities of cells were obtained by the shaking or aerating cultural method using media with a few ingredients [6,22] or a liquid medium in common use [28,30]. The same nutritional ingredients were not useful for semisolid or solid medium cultivation. In addition, by using the Thunberg method [20] and the Warburg apparatus [1,16], several substrates on the TCA cycle or compounds which could easily be introduced into the cycle were proven to have biochemical activity for C. fetus. However, a medium containing these substrates did not support the growth by static cultivation. Later, investigations of the nutritional requirements of this bacterium were made; however, it was difficult to determine a systematic nutritional requirement throughout these reports. Further, the research on nutritional requirements of this bacterium using the shake-culture method presented markedly different requirements for amino acids when compared with the