Day, Lawrence E., (Michigan State University, East Lansing) and Ralph N. Costilow. Physiology of the sporulation process in Clostridium botulinum. II. Maturation of forespores. J. Bacteriol. 88:695–701. 1964.—Clostridium botulinum, strain 62-A, did not sporulate endotrophically, but forespores matured to refractile, heat-resistant spores when replaced in solutions containing l-alanine and l-proline, l-isoleucine and l-proline, or l-alanine and l-arginine. Solutions of l-arginine or l-citrulline would not support the maturation process. Acetate, CO₂, and δ-amino valeric acid were produced during sporulation in a replacement solution of l-alanine and l-proline, indicating the operation of the Stickland reaction. There was no large uptake of either exogenous l-alanine or acetate during this process. Similarly, there was no apparent protein or nucleic acid synthesis, since high levels of chloramphenicol, 8-azaguanine, or mitomycin C failed to inhibit, and no significant amount of P³² was incorporated into the spore nucleic acids. Dipicolinic acid (DPA) was synthesized during forespore maturation. It is believed that these final steps in sporulation of C. botulinum require only an exogenous source of energy which can be obtained via the Stickland reaction system, and that the synthesis of DPA and other unknown materials relies primarily on endogenous substrates.