Terminals of olfactory bulb afferent (OB) and association (ASSN) fibers within the piriform cortex were characterized ultrastructurally. Identification was by electron microscopic (EM) autoradiography following injections of tritiated amino acids into the olfactory bulb and anterior piriform cortex. The results show that terminals of both fiber systems contain round vesicles and make asymmetrical synaptic contacts predominantly onto dendritic spines. Profiles with pleomorphic vesicles do not appear to be labeled from either site. Since there is strong evidence that both fiber systems generate excitatory postsynaptic potentials (EPSPs) in pyramidal cells, these results provide additional examples in the mammalian CNS of terminals with round vesicles and asymmetrical contacts that mediate an excitatory effect. Percentage density analysis and quantitative study of a large number of heavily labeled terminals revealed that while OB and ASSN terminals are similar in terms of vesicle shape and contact type, they differ in many morphological details including pre- and postsynaptic profile size, the packing density and distribution of synaptic vesicles, synaptic contact shape, and the presence of overlying neuroglial lamellae. However, large variations in appearance of different terminals of the same type are also present so that a small percentage of OB and ASSN terminals are indistinguishable morphologically in the absence of label. An important finding of the quantitative analysis is that spines contacted by lateral olfactory tract (LOT) terminals appear to be of two types based on a bimodal distribution in size and differences in morphology, while spines contacted by ASSN terminals appear to be of a single type. Comparison of these data with results from Golgi analysis indicates that ASSN terminals predominantly contact pyramidal cell spines while OB terminals contact both pyramidal and semilunar cell spines. Quantitative analysis of synaptic vesicles revealed that histograms of vesicle size for OB and ASSN terminals are virtually identical in shape, but peaks are slightly displaced (ASSN vesicles are 5% larger; significant with P less than .002). An analysis of the laminar distribution of OB and ASSN synaptic terminals revealed that while most OB terminals are segregated in layer Ia and most ASSN terminals in layer Ib, occasional OB terminals are observed up to approximately 50 micro deep to the Ia-Ib boundary and occasional ASSN terminals up to approximately 50 micro superficial to this boundary.