Microorganisms differ greatly in their minimal Aw requirement for growth, sporulation, and germination (Table 34.1). In general, molds and yeasts can grow at lower Aw values than bacteria; among pathogenic and spoilage bacteria, Gram-negatives require a slightly higher Aw than Gram-positives for growth. Sta. aureus, however,can grow at an Aw of 0.86. Sporulation by sporeforming bacteria occurs at Aw values in which the species and strains grow, whereas germination may occur at slightly lower Aw values. Toxin production may occur at the Aw of growth (by Clo. botulinum)or at a slightly higher than minimum Aw of growth (by Sta. aureus at 0.867 at 30ºC and 0.887 at 25ºC). The minimum Aw for microbial growth can vary with the type of solutes in a food. Clo. botulinum Type E failed to grow below an Aw of 0.97 when NaCl was used as a solute, but grew up to an Aw of 0.94 when glycerol was the solute. Pseudomonas fluorescens similarly showed a minimum Aw of 0.957 for growth with NaCl but 0.94 with glycerol. This is because glycerol enters freely inside the cell and thus does not cause osmotic stress as nonpermeable NaCl, sucrose,and similar solutes. Germination of spores in glycerol occurs at lower Aw because Clo. botulinum E spores, with glycerol, germinate at an Aw of 0.89, but do not grow below an Aw of 0.94. The growth rate of microorganisms also decreases as the Aw value is lowered. The growth rate of Sta. aureus reduces to about the 10% level at an Aw of 0.90 of its optimum growth rate at an Aw of 0.99.