In a previous paper we described the isolation and structure of hygrolidin (1), an antibiotic produced by Streptomyces hygroscopicus D-1166 which is specifically active against Valsa ceratosperma, the pathogen of apple canker disease. 1 belongs to a new group of 16-membered macrolide antibiotics. Recently, additional members of this group such as bafilomycins, a new macrolide substance and L-681,110 have been reported. Further screening for minor components of 1 resulted in the isolation of two metabolites named hygrolidin amide (2) and defumarylhygrolidin (3). In this paper, we wish to describe the isolation, structural elucidation and biological activities of these two minor components.